Increased B Regulatory Phenotype in Non-Metastatic Lymph Nodes of Node-Positive Breast Cancer Patients.

Tumour-draining lymph nodes (TDLNs) are centre in orchestrating the immune responses against cancer. The cellularity and lymphocyte subpopulations change in the process of cancer progression and lymph node involvement. B lymphocyte subsets and their function in breast cancer-draining lymph nodes have not been well elucidated. Here, we studied the influence of tumour metastasis on the frequencies of different B cell subsets including naïve and memory B cells as well as those which are known to be enriched in the regulatory pool in TDLNs of 30 patients with breast cancer. Lymphocytes were obtained from a fresh piece of each lymph node and stained for CD19 and other B cell-associated markers and subjected to flow cytometry. Our investigation revealed that metastatic TDLN showed a significant decrease in active, memory and class-switched B cells while the frequencies of B cells with regulatory phenotypes were not changed. However, CD27(hi) CD25(+) and CD1d(hi) CD5(+) B regulatory subsets significantly increased in non-metastatic lymph nodes (nMLNs) of node-positive patients compared with node-negative patients. Our data provided evidence that in breast cancer, metastasis of tumour to axillary lymph nodes altered B cell populations in favour of resting, inactive and unswitched phenotypes. We assume that the lymphatic involvement may cause an increase in a subset of regulatory B cells in non-metastatic lymph nodes.

Does the missing data imputation method affect the composition and performance of prognostic models?

BACKGROUND: We already showed the superiority of imputation of missing data (via Multivariable Imputation via Chained Equations (MICE) method) over exclusion of them; however, the methodology of MICE is complicated. Furthermore, easier imputation methods are available. The aim of this study was to compare them in terms of model composition and performance. METHODS: Three hundreds and ten breast cancer patients were recruited. Four approaches were applied to impute missing data. First we adopted an ad hoc method in which missing data for each variable was replaced by the median of observed values. Then 3 likelihood-based approaches were used. In the regression imputation, a regression model compared the variable with missing data to the rest of the variables. The regression equation was used to fill the missing data. The Expectation Maximum (E-M) algorithm was implemented in which missing data and regression parameters were estimated iteratively until convergence of regression parameters. Finally, the MICE method was applied. Models developed were compared in terms of variables significantly contributed to the multifactorial analysis, sensitivity and specificity. RESULTS: All candidate variables significantly contributed to the MICE model. However, grade of disease lost its effect in other three models. The MICE model showed the best performance followed by E-M model. CONCLUSION: Among imputation methods, final models were not the same, in terms of composition and perform-ance. Therefore, modern imputation methods are recommended to recover the information.

Survival of male breast cancer in fars, South of iran.

BACKGROUND: Although breast cancer in men is uncommon, its incidence rate has an increasing trend. Due to its low incidence, there are few studies in this subject and limited information is available. The purpose of this study was to investigate clinicopathological characteristics and survival of male breast cancer (MBC) in Fars Province, south of Iran. METHODS: The data for this study were obtained from the population based cancer registry of Vice-Chancellor for Health Affairs of Shiraz University of Medical Sciences and Shiraz hospitals between January 1, 1989 and January 1, 2008, including 64 patients with MBC. Demographic, clinical and pathological aspects were investigated. The Kaplan-Meier method was used for the determination of survival rate and Log Rank test for the comparison. The Cox proportional hazards model was used for the multiple analysis. RESULTS: The patients' mean age at the time of diagnosis was 60.3 years (SD=12.7). The most frequent age group (26.6%) was 51-60 years. The most common symptom (96.8%) was a palpable mass. The majority of patients (44.4%) had a symptom duration of less than or equal to 6 months. 56.3% of the patients had a tumor size of 2-4.9 cm. Forty six percent of the cases had axillary lymph node involvement. The median survival time was 10.0 years [95% confidence interval (CI): 6.0-14.0]. The 5 year overall survival rate was 66.0% (95% CI=51.0-81.0%). The median survival time of patients with axillary lymph node involvement was 8.2 years (95% CI=6.7-9.6) and for the cases without involvement was 12.0 years (95% CI=8.4-15.2). In addition to axillary lymph node involvement, positive family history in contrast to negative family history and left tumors in compari-son with right tumors were poorer prognostic factors in univariate analysis respectively (p=0.006, p=0.031). In multiple analysis, axillary lymph node involvement was an independent predictor of poorer survival (Hazard ratio=1.6, 95% CI=1.1-6.4, p=0.030) and the other variables did not have a significant effect. CONCLUSION: The mean age of MBC in this series is lower than that in western countries. It is compatible to the mean age of female breast cancer which is approximately one decade less than that in developed countries. The survival rate of MBC is relatively lower than that in western countries. Axillary lymph node involvement is an important prognostic factor in the survival of MBC. Multicenter population based studies with greater number of patients are required for better estimation of different aspects of MBC in Iran.

Multiple imputation in survival models: applied on breast cancer data.

BACKGROUND: Missing data is a common problem in cancer research. While simple methods such as completecase (C-C) analysis are commonly employed for handling this problem, several studies have shown that these methods led to biased estimates. We aim to address the methodological issues in development of a prognostic model with missing data. METHODS: Three hundred and ten breast cancer patients were enrolled. At first, patients with missing data on any of four candidate variables were omitted. Secondly, missing data were imputed 10 times. Cox regression model was fitted to the C-C and imputed data. Results were compared in terms of variables retained in the model, discrimination ability, and goodness of fit. RESULTS: Some variables lost their effect in complete-case analysis, due to loss in power, but reached significance level after imputation of missing data. Discrimination ability and goodness of fit of imputed data sets model was higher than that of complete-case model (C-index 76% versus 72%; Likelihood Ratio Test 51.19 versus 32.44). CONCLUSION: Our findings showed inappropriateness of ad hoc complete-case analysis. This approach led to loss in power and imprecise estimates. Application of multiple imputation techniques to avid such problems is recommended.

Association of interleukin-18 gene promoter polymorphisms with breast cancer.

Interleukin-18 [IL-18] gene promoter polymorphism is reported to be a genetic risk factor for several types of cancer. The aims of this investigation were to evaluate and compare the frequencies of IL-18 gene promoter polymorphisms at positions -137 [G/C] and -607 [C/A] in breast cancer patients and healthy controls as well as to study the contribution of these data with clinicopathological parameters at diagnosis. The studied populations comprised 250 cases with breast carcinoma and 206 healthy subjects. IL-18 gene promoter polymorphisms at positions -137 and -607 were amplified in patient and control groups using allele specific polymerase chain reaction [AS-PCR]. The frequencies of GG, GC and CC genotypes of -137 SNP were 141 [56.4%], 96 [38.4%] and 13 [5.2%] in patients vs. 110 [53.4%], 72 [34.9%] and 24 [11.7%] in controls, respectively. A significant decrease of the CC genotype was observed in patients [p = 0.04]. The frequency of the CC genotype at position -137 was also significantly higher in patients with metastasis than non-metastatic patients [21.4% vs. 4.3%] [p = 0.02]. There was no significant association between genotype frequencies at position -607 with breast cancer or its clinicopathological parameters at diagnosis. Moreover, allelic frequencies at these positions did not contribute to breast cancer incidence. The distribution of IL-18 gene haplotypes and genotype combinations were not significantly different between patients and normal control individuals. This is the first report investigating the contribution of IL-18 gene promoter polymorphisms to breast cancer. These results suggest contrast effects of IL-18 gene in cancer induction and progression. Key words: Breast cancer, IL-18, polymorphism.

Splenic lesions: FNA findings in 48 cases.

OBJECTIVE: To study fine needle aspiration (FNA) cytological findings of splenic lesions and assess the role of FNA in the diagnosis of splenomegaly or splenic tumours. METHODS: This study consisted of 48 cases, 25 males and 23 females. The ages ranged between 3 and 71 years. Most of these cases were aspirated under ultrasonographic guidance and a small number were also aspirated directly by using a 22- to 23-gauge needles. The smears were stained with Wright-Giemsa and Papanicolaou methods. Special stains were used whenever necessary. RESULTS: In this study 14 cases were diagnosed as lymphoma-leukaemia, 7 cases as tuberculosis, 12 cases as kala-azar, 2 cases as hydatid cyst, 3 cases as storage diseases, 3 cases as simple cyst, 2 cases as myeloproliferative disorders, 2 cases as malignant tumours and 3 cases as hamartomas (these were wrongly diagnosed as malignant tumours). CONCLUSION: Splenic aspiration is a safe procedure and is very useful in the diagnosis of parasitic and infectious diseases, especially in endemic countries like Iran.

Cytotoxic T lymphocyte antigen-4 promoter variants in breast cancer.

CTLA4 is a coinhibitory molecule expressed mainly on activated T lymphocytes. To test the putative involvement of CTLA-4 in inhibitory state of immunity to breast cancer, we genotyped 283 patients and 245 healthy control subjects for -1722 T/C, -1661 A/G, and -318 C/T single nucleotide polymorphisms in the promoter region of the CTLA4 gene. There were no significant differences in genotype, allele, or haplotype frequencies in all three loci between patients and healthy controls. Moreover, the incidence of the most frequent haplotype combination (TAC/TAC, T -1722, A -1661, C -318) was only slightly higher among healthy controls than patients (68.4 vs. 64.8%, P = 0.2). This haplotype combination was associated with lower stages of the disease (P = 0.0007), however, and higher estrogen receptor (ER) expression in patients (P = 0.006). Association with tumor prognostic or predictive factors was also observed with certain genotypes: the -1661 AA genotype was associated with lesser lymph node (LN) involvement (P = 0.017) and higher ER expression (P = 0.004), and the -318 CC genotype with lesser LN involvement (P = 0.007). These results suggest that CTLA4 promoter variants participate in the progression of breast cancer rather than in its initial development.

Insulin adsorbance to polyvinylchloride (PVC) surfaces of fluid container and infusion-set.

INTRODUCTION: In total parenteral nutrition (TPN) solution, adsorbance of insulin to polyvinylchloride (PVC) surfaces of fluid containers and infusion-sets, decrease the amount of insulin that reaches the patients. OBJECTIVE: To clarify the biding sites of insulin and to propose a solution to overcome this problem. METHODS AND MATERIALS: To each of four 1000ml. PVC bottles of 5 percent dextrose solution, 300 microunit of insulin per each milliliter of dextrose solution were added. Each bottle was then connected to an infusion-set and the system made to run at an infusion rate of 100ml. per hour. One milliliter samples were then collected from both the PVC bottles and infusion-sets-terminal, separately, immediately at the starting point (time zero) and 15, 30, 45 and 60 minutes thereafter. The concentrations of insulin were checked using insulin kits. RESULTS: At the starting point (time zero) the mean of insulin concentrations among four PVC bottles was 213.79 microunit per each milliliter of 5 percent dextrose solution. No significant fluctuation was noted in the concentration of insulin in the PVC bottles through 60th minute period. However the concentration of insulin at infusion-set- terminal decreased significantly at the end of the same hour (p. value = 0.004). CONCLUSION: Our results demonstrate that the adsorbance of insulin takes place at the surfaces of infusion sets. It follows therefore that increase in the primary dosage of insulin added to PVC infusion solutions and the selection of a suitable infusion set (polyethylene) seem to be beneficial for overcoming this problem.

Assessment of a breast cancer screening programme in Shiraz, Islamic Republic of Iran.

A breast cancer screening programme was evaluated for approximately 10,000 women aged 35 years and older. There were 67 cases of breast cancer. Highest rates of attendance were seen among younger women (35-44 years) and middle socioeconomic groups. Lowest rates were among those aged over 65 years and low socioeconomic groups. The rate of detection by self-examination was similar to that by health personnel examination. At all stages of screening, positive findings were most common among the high socioeconomic class. Attendance decreased steadily from first to last stages of serial screening. Although mammography is the most sensitive method of detection, because of its high cost we suggest establishing breast self-examination education programmes and encouraging women to self-examine.

Assessment of a breast cancer screening programme in Shiraz, Islamic Republic of Iran

A breast cancer screening programme was evaluated for approximately 10,000 women aged 35 years and older. There were 67 cases of breast cancer. Highest rates of attendance were seen among younger women [35-44 years] and middle socioeconomic groups. Lowest rates were among those aged over 65 years and low socioeconomic groups. The rate of detection by self-examination was similar to that by health personnel examination. At all stages of screening, positive findings were most common among the high socioeconomic class. Attendance decreased steadily from first to last stages of serial screening. Although mammography is the most sensitive method of detection, because of its high cost we suggest establishing breast self-examination education programmes and encouraging women to self-examine

Fine needle aspiration cytology of foreign bodies presenting as cystic abdominal masses. A report of three cases.

BACKGROUND: Foreign body material (gauze sponges) presented as cystic abdominal masses and were confused with malignant tumors. CASES: Two females and one male presented with abdominal masses. They had undergone laparotomy 5-12 years earlier. Clinically the masses were diagnosed as benign or malignant cystic lesions. Fine needle aspiration revealed necrotic material, hemosiderin-laden macrophages, foreign body giant cells, cholesterol crystals and many fragments of birefringent material. The possibility of malignancy was ruled out. Cut sections of the excised cystic lesions revealed gauze sponges surrounded by a thick, fibrotic wall. CONCLUSION: This report underscores the usefulness of fine needle aspiration in ruling out malignancy.

Bull's eye (target) inclusions in neoplastic cells in malignant serous effusions. A study of 289 cases.

OBJECTIVE: To study the prevalence and significance of bull's eye (target) inclusions in neoplastic cells in malignant serous effusions. STUDY DESIGN: We reviewed malignant pleural, peritoneal and pericardial effusions from 289 patients who had proven cancer at known primary sites. The ages of the patients ranged from 5 to 72 years; 166 were male and 123 female. RESULTS: Bull's eye inclusions are an uncommon finding and appeared in only 13 cases of metastatic adenocarcinoma of the breast, stomach, colon, lung, ovary, pancreas and urinary bladder. They were positively stained with periodic acid-Schiff stain with diastase. The inclusions were not seen in cells of nonadenocarcinomatous neoplasms, such as squamous cell carcinoma, oat cell (small cell) carcinoma, neuroblastoma, lymphoma and germ cell tumors. CONCLUSION: Bull's eye inclusions are found in about 5% of malignant serous effusions containing cells of metastatic adenocarcinoma. The primary site of an adenocarcinoma cannot be deduced on the basis of the presence of inclusions.

Reproductive behaviour in women in Shiraz, Islamic Republic of Iran.

Data on the reproductive behaviour of women in Shiraz are presented. A total of 9934 randomly selected women aged > or = 35 years of all socioeconomic levels were interviewed. The mean age at menarche +/- standard deviation was 13.96 +/- 1.23 years, mean age at first marriage was 17.10 +/- 4.24 years and mean age at first pregnancy was 19.50 +/- 3.90 years. The mean number of children was 4.56 +/- 1.70 children, although for illiterate women it was 6.76 +/- 1.76 children and for high-school- or university-educated women it was 3.36 +/- 1.70 children. The mean age at menopause was 47.80 +/- 3.78 years.

Papillary carcinoma of the breast. Cytologic study of nine cases.

OBJECTIVE: To study the cytologic findings of papillary breast carcinoma by fine needle aspiration. STUDY DESIGN: The study group consisted of fine needle aspiration (FNA) specimens of breast tumors from nine patients performed during the period 1988-1997. Eight were female, and one was male. The FNA results were compared with the final histologic diagnosis. RESULTS: The tumor sizes were 4-6.5 cm. The aspirations yielded a good amount of bloody material. The smears revealed high cellularity, papillary clusters, isolated low-to-tall columnar cells, mild to moderate atypia, hemorrhagic background, foam and hemosiderin-laden macrophages, calcification, rare mitoses, palisading row of cells and bipolar cytoplasmic eosinophilic granules. The smears were diagnosed as either suspicious or suggestive of papillary carcinoma. The histologic examination revealed invasive papillary carcinoma. CONCLUSION: Papillary carcinoma of the breast can be diagnosed by using a panel of cytologic findings that includes hypercellularity, papillary clusters, hemorrhagic background, palisading rows of tall columnar cells, cellular atypia and calcification. The interesting finding in this study was the presence of eosinophilic bipolar cytoplasmic granules, which has not been reported before.

Primary squamous cell carcinoma of the thyroid diagnosed by fine needle aspiration cytology. A report of two cases.

BACKGROUND: Primary squamous cell carcinoma of the thyroid is a rare malignant tumor that needs to be distinguished from other neoplasms. CASES: Two males aged 65 and 68 years presented with an enlarged right lobe of the thyroid. Thyroid scintigraphy revealed a cold nodule. Clinically malignancy was suspected. Fine needle aspiration (FNA) revealed numerous clusters and isolated malignant cells, dyskeratotic cells and deposits of eosinophilic granular keratin material. Subtotal thyroidectomy was performed. Histologic findings confirmed the cytologic diagnosis of squamous cell carcinoma. CONCLUSION: Primary squamous cell carcinoma of the thyroid is a rare tumor. FNA findings of this tumor were not reported before. FNA study cannot help to differentiate primary from metastatic squamous cell carcinoma of the thyroid.

Follicular variant of papillary carcinoma of the thyroid. A cytologic study of 15 cases.

OBJECTIVE: To study the cytologic findings of follicular variant of papillary thyroid carcinoma (FVPTC) and to compare them with the cytologic findings on other thyroid lesions. STUDY DESIGN: The study group consisted of aspirate smears from 15 cases of histologically proven FVPTC. The control group consisted of 152 cases, including adenomatous colloid goiter (70), usual papillary carcinoma (40), follicular adenoma (30), Hürthle cell neoplasm (7) and medullary carcinoma (5). RESULTS: The smears of FVPTC revealed numerous colloid balls in the background, multilayered microfollicles (rosettes), numerous nuclear grooves and inclusions in the monolayer sheets of follicular cells, very rare giant cells, absence of calcification and papillary clusters. Rosettelike microfollicles and numerous colloid balls were not seen in the control group. CONCLUSION: The combination of numerous colloid balls and rosettelike microfollicles was frequently seen in FVPTC. This combination was not observed in the control group.

Extramedullary plasmacytoma. Fine needle aspiration findings.

OBJECTIVE: To determine the role of fine needle aspiration cytology in the diagnosis of extramedullary plasmacytoma. STUDY DESIGN: The study group consisted of 13 patients with palpable masses at various sites. The tumors were aspirated for cytologic study. RESULTS: The smears revealed groups of mature and immature plasma cells at various stages of maturation. Mature plasma cells showed an eccentric nucleus and abundant, deep, basophilic cytoplasm with a paranuclear halo. Plasmablasts (immature plasma cells) showed a prominent, eccentric nucleus with single, large nucleolus and abundant, deep, basophilic cytoplasm with no paranuclear halo. Binucleate and multinucleate forms were also seen quite often. The tumors were excised, and the histologic sections confirmed the cytologic diagnosis. All the patients received radiotherapy. One patient (18 years old) developed recurrence and died due to extensive infiltration into the maxilla and mandible. Two patients (57 and 62 years) developed multiple myeloma one to two years after the excision of tumors, and both died two to three months later. The remaining 10 patients were alive and well at this writing. CONCLUSION: The smears from all 13 patients were diagnosed as extramedullary plasmacytomas by fine needle aspiration cytology.