Nuclear CK19-immunopositive pseudoinclusions as a new additional objective diagnostic feature of papillary thyroid carcinoma.

One of the key parameters in the diagnosis of papillary thyroid carcinoma (PTC) are true nuclear pseudoinclusions (NPs), which constitute invaginations of the cytoplasm into the nucleus. On the other hand, strong cytoplasmic expression of CK19 is a well-known attribute of PTC tumor cells. We analyzed NPs using CK19 immunohistochemistry in histological sections of 52 PTCs and seven noninvasive follicular thyroid neoplasms with papillary-like nuclear features (NIFTPs). Strong CK19+ NPs were present in 77% of PTCs, whereas NPs in hematoxylin and eosin (HE)-stained slides (HE NPs) were identified in only 48% of PTCs. Detection of CK19+ NPs enabled easier and objective recognition of NPs and better discrimination of NPs from pseudo-pseudoinclusions than detection of HE NPs. In the 15 of the 27 (55.5%) PTCs in which we could not discern HE NPs, strong CK19+ NPs could be identified reliably, quickly and easily. Moreover, all NIFTPs were negative for both CK19+ NPs and HE NPs. Detection of CK19+ NPs may refine the assessment of this important diagnostic feature and, hence, the microscopic diagnostic criteria of PTC. Thus, these findings may have implications for the accurate diagnosis of PTC and NIFTP.

BRCA1 and PARP1 mRNA expression during progression from normal breast to ductal carcinoma in situ and invasive breast cancer: a laser microdissection study.

The contribution of DNA damage repair mechanisms to the progression of normal breast to ductal carcinoma in situ (DCIS) and invasive ductal carcinoma is largely unknown. The purpose of this report was to assess the mRNA expression levels of two important genes associated with DNA repair, BRCA1 and PARP1, in normal breast tissue, DCIS G1, G2 and G3, and co-existing adjacent invasive ductal carcinoma. BRCA1 and PARP1 mRNA expression was assessed in 32 ductal carcinomas in situ of the breast using a laser microdissection and pressure catapulting system and quantitative real-time PCR. The relative expression of BRCA1 mRNA was significantly increased in DCIS G2 and DCIS G3 relative to normal breast tissue (p = 0.02, p = 0.001, respectively). Significant differences in BRCA1 expression were observed between DCIS G1 and G2 (p = 0.02) and between DCIS G1 and G3 (p = 0.0007). No significant differences in BRCA1 expression were observed between normal breast tissue and DCIS G1 and between DCIS component and adjacent invasive ductal carcinoma. No significant differences in the relative expression of PARP1 mRNA were observed between groups. Increased BRCA1 mRNA expression (but not PARP1 mRNA) occurs early in the development of breast cancer, i.e. at the noninvasive (DCIS) stage, suggesting a demand for increased activity of a DNA double-strand break repair by homologous recombination. DCIS G1 and normal breast tissue share highly similar BRCA1 and PARP1 expression level. This finding supports the idea that DCIS G1 belongs to a separate family of precursor lesions with low malignant potential.

Different CHEK2 germline mutations are associated with distinct immunophenotypic molecular subtypes of breast cancer.

Germline mutations in BRCA1 were already linked to basal-like subtype of immunophenotypic molecular classification of breast cancer (BC). However, it is not known whether mutations in other BC susceptibility genes are associated with molecular subtypes of this cancer. We tested the hypothesis that distinct mutations in another BC susceptibility gene involved in DNA repair, i.e., CHEK2 may be associated with particular immunophenotypic molecular subtypes of this cancer. Two groups of patients: 1255 with BCs and 5496 healthy controls were genotyped for four CHEK2 mutations (I157T and three truncating mutations: 1100delC, IVS2 + 1G > A, del5395). BCs were tested by immunohistochemistry on tissue microarrays for ER, PR, HER-2, EGFR, and CK5/6 and were assigned to appropriate subtypes of immunophenotypic molecular classification. There was a significant association between CHEK2 mutations and the immunophenotypic molecular classification (P = 0.004). CHEK2-associated cancers were predominantly luminal (108/117 = 92.3%). CHEK2-I157T variant was associated with the luminal A subtype (P = 0.01), whereas CHEK2-truncating mutations were associated with the luminal B subtype (P = 0.005). Comparing the prevalence of CHEK2 mutations in BC with controls revealed that carriers of an I157T variant had OR of 1.80 for luminal A subtype and carriers of truncating mutations had OR of 6.26 for luminal B subtype of BC. To our knowledge, this is the first study showing that specific mutations in the same susceptibility gene are associated with different immunophenotypic molecular subtypes of BC. This association represents independent evidence supporting the biological significance of immunophenotypic molecular classification of BC.

KRAS mutation testing in colorectal cancer as an example of the pathologist's role in personalized targeted therapy: a practical approach.

Identifying targets for personalized targeted therapy is the pathologist's domain and a treasure. For decades, pathologists have had to learn, understand, adopt and implement many new laboratory techniques as they arrived on the scene. Pathologists successfully integrate the results of those tests into final pathology reports that were, and still are, the basis of clinical therapeutic decisions. The molecular methods are different but no more difficult to comprehend in the era of "kit procedures". In recent years, the development of targeted therapies has influenced routine practices in pathology laboratories because the use of molecular techniques is required to include clinically useful predictive information in the pathology report. Pathologists have the knowledge and expertise to identify particular gene mutations using the appropriate molecular tests currently available. This review focuses on the most important recent developments in KRAS mutation testing in metastatic colorectal cancer (CRC), and shows that a pathologist is involved in 10 stages of this procedure. Recent studies have shown that highly sensitive, simple, reliable and rapid assays may significantly improve the identification of CRC patients resistant to anti-EGFR therapy. Thus, direct sequencing does not seem to be an optimal procedure of KRAS testing for clinical purposes. Twelve currently available high-sensitivity diagnostic assays (with the CE-IVD mark) for KRAS mutation testing are briefly described and compared. The suggested pathology report content for somatic mutation tests is described. In conclusion, evidence is presented that sending away paraffin blocks with tumor tissue for KRAS mutation testing may not be in the best interest of patients. Instead, an evidence-based approach indicates that KRAS mutation testing should be performed in pathology departments, only with the use of CE-IVD/FDA-approved KRAS tests, and with the obligatory, periodic participation in the KRAS EQA scheme organized by the European Society of Pathology as an independent international body.

PARP-1 expression in breast cancer including BRCA1-associated, triple negative and basal-like tumors: possible implications for PARP-1 inhibitor therapy.

Despite ongoing trials of PARP inhibitors in the treatment of breast cancer (BC), the extent of poly(ADP-ribose)polymerase-1 (PARP-1) protein expression in BCs, which may influence treatment results, is not known. The purpose of this report is to assess expression of PARP-1 in BC including BRCA1-associated, triple negative (TN), and basal-like tumors. Immunohistochemistry with a PARP-1 antibody on tissue microarrays from 130 BRCA1-associated and 594 BRCA1-non-related BCs was used. The vast majority of breast carcinomas expressed high level of nuclear PARP-1 protein and a small percentage of tumors exhibited both nuclear and cytoplasmic PARP-1 expression. There was a significant difference between the mean nuclear PARP-1 quickscore in BRCA1-associated versus BRCA1-non-associated carcinomas in all tumors (P < 0.0001), in the basal-like group (P = 0.0086), TN (P = 0.0015), and non-basal-like groups (P = 0.016) but not in the non-TN group. Among BRCA1-associated BCs, low PARP-1 expression was found in 18.5% of all cases, 18.9% of basal-like and 21% of TN cancers. Among BRCA1-non-related tumors, low PARP-1 expression was found in 8.8% of all cases, 3.1% of basal-like, and 2.7% of TN cancers. PARP-1 expression is significantly associated with BRCA1 status in basal-like and TN BCs. The assessment of PARP-1 expression in tumor samples may improve the selection of BC patients for PARP inhibitor therapy.

Morphometric distinction of signet-ring cell adenocarcinoma cells from foamy macrophages in gastric endoscopic biopsies.

AIMS: To distinguish signet-ring cancer cells from foamy macrophages in the small gastric endoscopic biopsies using objective morphometric measurements of nuclei. MATERIAL AND METHODS: Using computerized image analysis, the mean nuclear area, length, breadth, perimeter and roundness were analyzed in histological sections of ten gastric endoscopic biopsies with signet-ring cell adenocarcinoma and four benign lesions with numerous foamy macrophages. RESULTS: Nuclei of signet-ring cell adenocarcinoma were significantly bigger than nuclei of foamy macrophages. Mean nuclear area (34.25 µm2 for carcinoma cells vs. 25.41 µm2 for macrophages) and mean nuclear breadth (5.82 µm vs. 4.99 µm, respectively) differed significantly (P < 0.05), whereas the remaining parameters did not. CONCLUSION: Nuclear morphometry can distinguish foamy macrophages from signet-ring cell adenocarcinoma cells in endoscopic gastric biopsies.

Reed-Sternberg cells in classical Hodgkin lymphoma in children seem to be predominantly oestrogen receptor α negative and oestrogen receptor ß positive.

Oestrogen receptor α (ERα) is responsible for activation of gene transcription, while oestrogen receptor ß (ERß) serves as a negative regulator of ERα function. Since ER status is a prognostic and predictive factor in some cancers, we analysed the immunohistochemical expression of ERα and ERß in Reed-Sternberg (RS) cells in paraffin-embedded lymph node specimens from 27 children with classical Hodgkin lymphoma (HL) in relation to histological type, clinical stage, age, and gender. Percentage of RS cells with positive nuclear reaction for the presence of ERα and/or ERß was assessed. ERα positive RS cells were present in 11% (3/27) of lymph nodes (range 1-8%, mean 0.4%) whereas ERß positive RS cells were detected in 96% (26/27) of lymph nodes (range 1-97.5%, mean 61.8%). The highest percentage of ERß positive RS cells was observed in patients with the most advanced (IVB) disease as compared to patients with lower stages (90.3% vs. 56.9% respectively, p = 0.004). To the best of our knowledge this is the first report on the expression of ERß in RS cells in children. We conclude that RS cells in classical HL in children seem to be mainly ERß positive and ERα negative.

Activation of NF-ĸB in leukemic cells in response to initial prednisone therapy in children with acute lymphoblastic leukaemia: relation to other prognostic factors.

Nuclear factor ĸB (NF-ĸB) is a transcription regulator of proliferation and cell death. Increased activation of NF-ĸB may be responsible for treatment failure in children with acute lymphoblastic leukaemia (ALL). This study aimed to assess changes in NF-ĸB activation in peripheral blood mononuclear cells prior to and after 6 and 12 h of prednisone administration in relation to age, initial WBC count at diagnosis and early treatment response in childhood ALL. The study comprised 55 children with de novo ALL. Cells were stained with mouse anti-NF-ĸB (p65) antibody followed by goat anti-mouse antibody conjugated with FITC and measured by laser scanning cytometer. The nuclear/cytoplasmic (N/C) ratio of NF-ĸB reflecting activation of NF-ĸB was decreased 12 h after treatment in the standard risk group patients, whereas it remained statistically unchanged in the non-standard risk group patients. Changes in the N/C ratio of NF-ĸB were not associated with age and early treatment response; however, in children with an initial WBC count higher than 20 000/µl at diagnosis, this ratio was increased after 6 and 12 h from prednisone administration. The association of higher activation of NF-ĸB with an elevated initial WBC count suggests that activation of NF-ĸB may be responsible for treatment failure in children with ALL.

Mutational analysis of CDKN2A gene in a group of 390 larynx cancer patients.

CDKN2A gene belongs to the genes involved in cell cycle regulation. When is absent or inactivated by mutation or promoter hypermethylation a cell may undertake an uncontrolled proliferation. Inactivation of CDKN2A gene is observed in many human malignancies, including larynx cancer. In this study we investigated mutations in exon 1 and exon 2 of CDKN2A gene in a large group of 390 laryngeal cancers. We found 40 different alterations (17%) and nearly half of them was not described previously. Out of these alterations two transversions in codon 108: c.322G>C (Asp108His) and c.322G>T (Asp108Tyr) as well as a G>A transition in codon 110 (Trp110X) were found more frequently (altogether: 7 cases in codon 108 and 10 cases in codon 110). This result, concerning the location of these codons in the ankyrin repeat structures, may suggest that these two codons may be critical hot-spots in larynx carcinogenesis.

Effects of recombinant human growth hormone (rhGH) replacement therapy on detailed immunologic parameters in somatotropine--deficient paediatrics patients prior and after 6 months of rhGH treatment.

OBJECTIVE: This study aims at assessing how recombinant human growth hormone treatment of children and young people suffering from isolated growth hormone deficiency affects some selected parameters of the immune system: a percentage of lymphocytes, granulocytes, monocytes, concentrations of A, G, and M immunoglobulins, a percentage of T lymphocytes divided into subpopulations CD4 and CD8, a percentage of NK and B lymphocytes, and phagocytic activity of granulocytes and monocytes. MATERIALS AND METHODS: The study comprised 30 children and young people aged 4.2-18 years with isolated growth hormone deficiency both prior to and 6 months after rhGH (recombinant human growth hormone) treatment with a dose of 0.093 IU/kg every 24 hr. The control group comprised 25 healthy children with normal height in the respective age bracket. Labelling was conducted by flow cytometry FACS manufactured by Becton-Dickinson using both labelled antibodies and PHAGOTEST® commercial kit (Orpegen). Concentrations of A, G and M immunoglobulins in blood serum were assessed by means of immunoturbidimetric method using COBAS (manufactured by Roche). RESULTS: The lowest percentage of active granulocytes in PHAGOTEST® was found in a group examined prior to treatment compared to the control group. The percentage increased after 6 months of rhGH treatment to values comparable with the control group. Although mean concentrations of IgM and IgA after 6 months of treatment with rhGH significantly decreased in comparison with those determined prior to treatment, they still remained within the baseline norm. No significant differences in the phagocytic activity of monocytes, IgG concentration, % of NK lymphocytes, T lymphocytes divided into CD4 and CD8 lymphocytes, B lymphocytes and CD4/CD8 lymphocytic index were found. None of the patients exhibited any clinical symptoms of immune disorders. CONCLUSION: rhGH treatment of patients with isolated growth hormone deficiency can have positive influence on the phagocytic activity of scavenger cells, mainly granulocytes, which in children with isolated growth hormone deficiency seems to be lower than in their health peers. Growth hormone treatment of children with isolated growth hormone deficiency does not significantly affect the activity of the immune system expressed by the phagocytic activity of monocytes, the percentage of B, T and NK lymphocytes and IgG concentration in blood serum.

Simultaneous assessment of p53 and MDM2 expression in leukemic cells in response to initial prednisone therapy in children with acute lymphoblastic leukemia.

Ineffective apoptosis is one of main causes of a treatment failure in childhood acute lymphoblastic leukemia (ALL). p53 plays a crucial role in triggering apoptosis of ALL in response to prednisone treatment. MDM2 is the endogenous inhibitor of apoptosis that downregulates the functional activity of p53 protein. This study is aimed to evaluate changes in MDM2 and p53 expression in peripheral blood mononuclear cells collected from children with ALL prior to and after 6 and 12 h of prednisone administration in relation to early treatment response. The study comprised 35 children with newly diagnosed ALL, subdivided into good (n = 24) and poor (n = 11) early treatment responders. MDM2 - associated APC fluorescence and p53 - associated FITC fluorescence were measured by the laser scanning cytometer. In the group of poor responders, p53 and MDM2 fluorescence were significantly higher than in the group of good responders. In the group of good early treatment responders, a statistically significant rise of p53 fluorescence measured in the nucleus and in the cytoplasm 12 h after prednisone administration as well as increase in MDM2 fluorescence measured in the cytoplasm 6 and 12 h after prednisone administration were seen. These data suggest that pretreatment overexpression of MDM2 protein may contribute to poor early treatment response.

Genetic contribution to all cancers: the first demonstration using the model of breast cancers from Poland stratified by age at diagnosis and tumour pathology.

The aim of the study is to verify the hypothesis that genetic polymorphisms are associated with the predisposition to all malignancies. Using as a model breast cancers from the homogenous Polish population (West Pomeranian region) after stratification of 977 patients by age at diagnosis (under 51 years and above 50 years) and by tumour pathology (ductal cancers--low and high grade, lobular cancers, ER-positive/negative) we tested this hypothesis. Altogether 20 different groups of breast cancer cases have been analyzed. The results were compared to a group of unaffected controls that were matched by age, sex, ethnicity and geographical location and originated from families without cancers of any site among relatives. Molecular alterations selected for analyses included those which have been previously recognized as being associated with breast cancer predisposition. Statistically significant differences between the breast cancer cases and controls were observed in 19 of the 20 analyzed groups. Genetic changes were present in more than 90% of the breast cancer patients in 18 of 20 groups. The highest proportion of cases with constitutional changes-99.3% (139/140) was observed for lobular cancers. The number and type of genetic marker and/or the level of their association with the specific cancer predisposition was different between groups. Markers associated with majority of groups included: BRCA1, CHEK2, p53, TNRnTT, FGFRnAA, XPD CC/AA and XPD GG. Some markers appeared to be group specific and included polymorphisms in CDKN2A, CYP1B1, M3K nAA, and RS67.

Good early treatment response in childhood acute lymphoblastic leukemia is associated with Bax nuclear accumulation and PARP cleavage.

BACKGROUND: Bax activation and intracellular redistribution as well as its prognostic significance during steroid-induced apoptosis of leukemic cells in childhood acute lymphoblastic leukemia (ALL) remain a matter of controversy. The purpose of this study was to assess time-resolved changes in Bax activation and its intracellular distribution as well as the percentage of apoptotic cells evaluated by PARP cleavage in response to prednisone treatment in childhood ALL. MATERIAL/METHODS: The study comprised 43 children with de novo ALL. Bax activation and PARP cleavage were measured by laser scanning cytometry in peripheral blood mononuclear cells collected prior to and 6 and 12 hours after prednisone administration. RESULTS: The mean pretreatment proportion of p89 PARP-positive cells was 3.5%. Six and twelf hours after prednisone administration it increased significantly (p<0.01) only in the group of good treatment responders. A significant rise (p<0.05) in cytoplasmic Bax expression was seen in the good responders as early as 6 hours after prednisone administration followed by a significant rise in Bax nuclear expression after 12 hours. At the same time points the mean percentage of apoptotic cells as well as Bax expression in the cytoplasm and nucleus remained unchanged in the group of poor responders. CONCLUSIONS: Increased Bax nuclear accumulation (and possibly also aggregation) together with increased PARP cleavage observed within 12 hours after prednisone oral administration were associated with and may predict good outcome in children with newly diagnosed ALL.

Prognostic significance of morphometric parameters of nucleoli and nuclei of invasive ductal breast carcinomas.

The aim of this study was to evaluate associations between seven morphometric parameters of the nucleoli and nuclei of methyl green and pyronin Y (MG-PY) stained tumour cells of invasive ductal breast carcinoma with relapse-free survival (RFS) and overall survival (OS) time. Histological sections from 150 invasive ductal breast cancers were stained with MG-PY and the following parameters were evaluated by computer image analysis: the nucleolar area, long to short nucleolar axis ratio, nucleolar shape parameter assessing the degree of nucleolar roundness, long to short nuclear axis ratio, number of nucleoli in the nucleus and the percentage of the nuclear cross-section surface area occupied by the nucleoli. A statistically significant association between a nucleolar shape polymorphism and the number of nucleoli in the nuclei of tumour cells and the RFS but not OS was found in the entire group of patients as well as patients with axillary lymph node metastases. A higher polymorphism of nucleolar shape and a higher number of nucleoli in the nuclei of breast cancer cells were associated with decreased relapse-free survival (p < 0.05). The remaining morphometric parameters showed no statistically significant association with RFS or OS. The results indicate that morphometry of nucleoli in MG-PY stained histological sections can be useful in the analysis of associations between nucleolar parameters and prognosis of patients with invasive breast cancer.

Fatty acids distribution and content in oral squamous cell carcinoma tissue and its adjacent microenvironment.

Squamous cell carcinoma of the oral cavity mucosa grows under conditions of poor oxygenation and nutrient scarcity. Reprogramming of lipid biosynthesis accompanies tumor growth, but the conditions under which it occurs are not fully understood. The fatty acid content of the serum, tumor tissue and adjacent tumor microenvironment was measured by gas chromatography in 30 patients with squamous cell carcinoma grade 1-3. Twenty-five fatty acids were identified; their frequencies and percentages in each of the environments were assessed. Nineteen of the twenty-five fatty acids were found in tumor tissue, tumor adjacent tissue and blood serum. Of them, 8 were found in all thirty patients. Percentages of C16:0 and C18:1n9 were highest in the tumor, C18:1n9 and C16:0 were highest in tumor adjacent tissue, and C16:0 and C18:0 were highest in blood serum. The frequencies and amounts of C22:1n13, C22:4n6, C22:5n3 and C24:1 in tumor adjacent tissues were higher than those in blood serum, independent of the tumor grade. The correlations between the amount of fatty acid and tumor grade were the strongest in tumor adjacent tissues. The correlations between particular fatty acids were most prevalent for grade 1+2 tumors and were strongest for grade 3 tumors. In the adjacent tumor microenvironment, lipogenesis was controlled by C22:6w3. In blood serum, C18:1trans11 limited the synthesis of long-chain fatty acids. Our research reveals intensive lipid changes in oral cavity SCC adjacent to the tumor microenvironment and blood serum of the patients. Increase in percentage of some of the FAs in the path: blood serum-tumor adjacent microenvironment-tumor, and it is dependent on tumor grade. This dependency is the most visible in the tumor adjacent environment.

In vitro assessment of adhesion molecules expression by human endothelial cells cocultured with c-erbB2-positive and c-erbB2-negative breast carcinoma cell lines.

OBJECTIVE: In vitro assessment of ICAM-1, VCAM-1 and P-selectin expression on endothelial cells cocultured with c-erbB2-positive (SK-BR-3) and c-erbB2-negative (MCF-7) breast carcinoma cell lines. MATERIAL AND METHODS: The expression of adhesion molecules was determined by immunofluorescence, and measured by a laser scanning cytometer. RESULTS: Endothelial cells cocultured with c-erbB2-negative breast carcinoma cell line--as compared to endothelial cells cultured without carcinoma cells--demonstrated an increase in the expression of VCAM-1 and P-selectin, a decrease in the expression of ICAM-1, and an increase of the maximal pixel intensity of ICAM-1, VCAM-1 and P-selectin orange fluorescence. A higher increase of the expression and maximal pixel intensity of ICAM-1, VCAM-1 and P-selectin was observed in endothelial cells cocultured with c-erbB2-positive breast carcinoma cells as compared to endothelial cells cocultured with c-erbB2-negative cell line. CONCLUSIONS: An increase in the expression of adhesion molecules on endothelial cells cocultured with c-erbB2-positive breast carcinoma cells may be responsible for the increased adhesion of tumor cells to endothelium and the increased invasiveness of c-erbB2-positive breast carcinomas.

Intranuclear nucleolin distribution during cell cycle progression in human invasive ductal breast carcinomas in relation to estrogen receptor status.

BACKGROUND: Nucleolin (NU) can be found in the nucleus in aggregates and in a diffused form. The expression and distribution of these two intranuclear pools of nucleolin was compared in estrogen receptor (ER)-positive and -negative invasive ductal breast carcinomas in relation to cell cycle phases. MATERIALS AND METHODS: Using a laser scanning cytometer NU was measured within the whole nucleus, within nucleolin aggregates (NUA) and within the NUA-free karyoplasm. The G1-, S- and G2M-phase cell subpopulations were distinguished on DNA histograms. RESULTS: The nuclei of ER-positive and ER-negative breast carcinomas in the G2M-phase differed in the expression of NU in the NUA and in the NUA-free karyoplasm, whereas in the S-phase they only differed in the NUA-free karyoplasm expression. In ER-positive carcinomas NU levels rose from G1 to G2M in both the NUA and the NUA-free karyoplasm, whereas in ER-negative carcinomas the NU levels increased only between G1/S in the NUA and between S/G2M in the NUA-free karyoplasm. CONCLUSION: There are significant differences in nucleolin expression and localization during cell cycle progression depending on ER status.

Immunohistochemical analysis of thymidylate synthase expression in gastric carcinoma: correlation with clinicopathological parameters and survival.

The correlation of thymidylate synthase (TS) expression in gastric cancers with tumor histology and prognostic or predictive information remains unclear. Most studies have involved Asian populations, with few conducted in European cohorts. Moreover, all published studies analyze TS expression using semi-quantitative methods. This retrospective study evaluated the association of TS expression in tumor cells with gastric carcinoma histological type, with selected clinicopathological parameters, and with the prognosis of patients who underwent surgical treatment. TS expression was detected using immunochemistry and objectively assessed by computerized image analysis of tumor cells in 100 gastric cancers. We found that high TS expression was significantly more common in intestinal than in diffuse type of gastric cancer according to Lauren classification (P=0.0003); in type I carcinomas compared to type IV according to Goseki classification (P=0.002); and in gastric cancers in men than women (P=0.04). Low TS expression was found more often in carcinomas in the middle and lower third of the stomach than in cancers in the upper third of the stomach (P=0.009 and P=0.001, respectively). In the subgroup of 25 patients without lymph node metastases (stage I+II), high TS expression was associated with better DFS (83% for high TS expression versus 38,5% for low TS expression, P=0.03). The results (1) indicate significant correlation between the Lauren and Goseki histopathological classifications of gastric cancer and TS expression in tumor cells, (2) suggest that high TS expression may be a positive prognostic marker with regard to DFS in patients with gastric cancer without involvement of regional lymph nodes who underwent radical surgical treatment and were not treated with preoperative chemotherapy. Prognostic results need confirmation in larger cohorts.

The NOD2 3020insC mutation and the risk of colorectal cancer.

Several predispositions to colorectal cancer have been identified, but little is known about genetic susceptibilities to disease in older persons. Colorectal cancer is a risk in Crohn's disease and is believed to be associated with an inappropriate inflammatory response. Recently, the NOD2 gene has been associated with Crohn's disease, which further strengthens the notion that the inflammatory response plays a crucial role in this disease. Several mutations have been identified in the NOD2 gene, which appear with significantly higher frequency in patients with the disease. One such mutation (3020insC) is believed to be clearly causative because it results in a prematurely truncated protein with a predicted reduction in functional efficiency. In this report, we have examined the frequency of the 3020insC mutation in a series of 856 individuals including 556 patients with colorectal cancer. The frequency of the 3020insC mutation in a consecutive series of 250 non-hereditary nonpolyposis colorectal cancer patients >50 years of age was significantly elevated compared with the control population (odds ratio, 2.23; P = 0.0046). The results indicate that NOD2 may be a predisposing factor to colorectal cancer characterized by an older average age of disease onset in persons who do not harbor any other genetic predisposition to disease.