PI3K/AKT Signaling in Breast Cancer Molecular Subtyping and Lymph Node Involvement.

Lymph node metastatic involvement persists to be among the most important predictors of recurrence and survival in breast carcinoma (BC). This study is aimed at investigating possible gene expression differences in primary BC between patients with or without lymph node involvement at the time of diagnosis. In a retrospective study, we investigated the potential prognostic role of 9 candidate biomarkers at the mRNA level in a cohort of 305 breast cancer patients, 151 lymph node-negative (LN-) and 154 lymph node-positive (LN+) individuals. The analyzed genes belonged to the RAS pathway (RAF1, ERBB2, PIK3CB, AKT1, AKT2, and AKT3), RB pathway (RB1 and CDK2), and cellular differentiation (KRT8). Their expression profiles were investigated by RT-qPCR and were correlated to immunohistochemically based molecular subtypes and BC clinical and pathological features. The differential expression of several genes in the primary tumor tissue was related to the LN involvement. Some of those genes, including PIK3CB, RB1, and AKT3, were more expressed in LN- BC patients, while some others, notably ERBB2 and AKT1, in LN+ ones. Among the candidate biomarkers, the expression levels of AKT isoforms influenced also patients' survival rates. In detail, higher expression levels of AKT1 and AKT2 negatively influenced overall patients' survival, and in particular, AKT2 expression levels defined a group of luminal B BC patients with shorter cancer-specific survival. On the contrary, longer cancer-specific survival was recorded in luminal A BC patients with higher expression levels of AKT3. That finding was also confirmed by Cox multivariate analysis. The same AKT3 resulted to be a possible candidate predictive biomarker for Tamoxifen response. In conclusion, our study highlighted the complex regulation of the PI3K/AKT pathway in BC and its differences in BC patients with and without lymph node involvement.

Prevalence of intestinal metaplasia in the distal oesophagus, oesophagogastric junction and gastric cardia in symptomatic patients in north-east Italy: a prospective, descriptive survey. The Italian Ulcer Study Group "GISU".

BACKGROUND: Incidence of adenocarcinoma of distal oesophagus and gastric cardia, probably arising from areas of intestinal metaplasia, has been increasing rapidly. AIMS: To define prevalence of intestinal metaplasia of distal oesophagus, oesophagogastric junction and gastric cardia and to evaluate potential associated factors, by means of a prospective multicentre study including University and teaching hospitals, and primary and tertiary care centres. PATIENTS: Each of 24 institutions involved in study enrolled 10 consecutive patients undergoing first-time routine endoscopy for dyspeptic symptoms. METHODS: Patients answered symptom questionnaires and underwent gastroscopy Three biopsies were taken from distal oesophagus, oesophago-gastric junction and gastric cardia, and were stained with haematoxylin and eosin. Specimens were also evaluated for Helicobacter pylori infection. RESULTS: A total of 240 patients (124 male, 116 female; median age 56 years, range 20-90) were enrolled in study. Intestinal metaplasia affected distal oesophagus in 5, oesophago-gastric junction in 19 and gastric cardia in 10 patients. Low-grade dysplasia was found at distal oesophagus and/or oesophago-gastric junction of 3/24 patients with intestinal metaplasia vs 2/216 without intestinal metaplasia (p<0.05). A significant association was found between symptoms and presence of intestinal metaplasia, regardless of location, and between Helicobacter pylori infection and intestinal metaplasia at oesophago-gastric junction. CONCLUSIONS: Intestinal metaplasia of distal oesophagus, oesophagogastric-junction and gastric cardia is found in a significant proportion of symptomatic patients undergoing gastroscopy and is associated with dysplasia in many cases. Although prevalence of dysplasia seems to decrease when specialized columnar epithelium is found in short segment, or even focally in oesophago-gastric junction, these small foci of intestinal metaplastic cells may represent source of most adenocarcinomas of cardia.

Angiomyolipoma of the parotid gland: a case report.

Angiomyolipoma is a hamartomatous process that most frequently occurs as a single lesion or multiple foci in the kidneys of patients affected by tuberous sclerosis. Angiomyolipoma can also arise in extrarenal sites, among which the liver is the most frequently recorded. Only rare cases of angiomyolipoma located in the head and neck region (ear and oral and nasal cavity) have been described. The purpose of the present article is to report a case of angiomyolipoma of the parotid gland. A 68-year-old woman appeared for treatment with a slow-growing nodule located in her right parotid gland. Ultrasound examination revealed a heterogeneous nodule with well-defined margins. The nodule was surgically removed by total parotidectomy and showed the characteristic appearance of angiomyolipoma, with an admixture of fat smooth muscle cells, and tortuous, thick-walled blood vessels. Careful physical examination of the patient failed to reveal features of tuberous sclerosis. Angiomyolipoma should be considered in the differential diagnosis of mesenchymal lesions involving the salivary gland.

Immunoglobulin heavy chain diversity genes rearrangement pattern indicates that MALT-type gastric lymphoma B cells have undergone an antigen selection process.

Gastric MALT lymphoma usually develops from chronic gastritis, the vast majority of which (>90%) is associated with Helicobacter pylori infection. We sequenced the third complementarity determining region (CDR3) of immunoglobulin heavy chain genes in 19 gastric MALT lymphoma clones to determine the pattern of variable (V), diversity (D) and joining (J) gene utilization during immunoglobulin gene rearrangement. DNA was extracted from paraffin-embedded sections and the rearranged CDR3 regions were amplified using a semi-nested polymerase chain reaction (with primers complementary to the conserved framework-three segment of the variable region [FR3A] and J regions). The DNA used for cloning and sequencing was obtained after purification of monoclonal bands excised from polyacrylamide gels. The N-D-N region specific to each clone was compared with known germline D sequences. Similarly to that observed in normal and leukaemic B cells, our series of gastric MALT lymphomas showed apparent preferential utilization of genes from the DXP family. In two cases no similarity between the CDR3 nucleotide sequences of the neoplastic clones and the known germline D sequences could be found. In 10/19 analysed alleles the lymphoma B-cell clones appeared to contain two D gene segments (D-D recombination), a rare occurrence in normal individuals but one which has been described as a significant event in the determination of idiotype expression and antigen-binding affinity. Remarkably, despite the use of different D and J segments, the resultant amino acid sequences matched in two patients, suggesting the presence of a common selecting antigen. The observed pattern of D gene rearrangement suggests that MALT lymphoma B-cell clones have undergone antigen selection, which seems to indicate that the antigen stimulation plays a pivotal role in the development of the lymphoma.

Role of fine-needle aspiration cytology in the preoperative evaluation of smooth muscle tumors.

A preliminary study was undertaken to assess the feasibility and the diagnostic role of fine-needle aspiration cytology (FNAC) in the preoperative evaluation of eight uterine smooth muscle tumors manifesting as single large masses with signs of growth. Percutaneous FNAC was performed under echographic control with a 22-gauge needle and the material was stained according to conventional techniques. Histology of surgically resected specimens was available for final diagnosis and comparative analysis in all the cases, including five leiomyomas (LM), one smooth muscle tumor of uncertain malignant potential (TUMP), and two low-grade leiomyosarcomas (LMS). Cellularity, as indicated by the density (crowding) of nuclei reflecting the amount of cytoplasmic volume, and the cohesiveness of the tissue fragments in the smears appeared to be the most important diagnostic parameters in the distinction between LM and LMS. LM usually showed few scattered poorly cellular fragments of highly cohesive tapering cells without nuclear crowding and with abundant cytoplasm. LMS usually showed a large number of single cells and fragments of loosely arranged tapering cells with nuclear enlargement and crowding and ill-defined scanty cytoplasm. Borderline forms such as TUMP were hardly distinguishable from LMS and LM. FNAC appears to be a feasible preoperative procedure in uterine smooth muscle tumors and may play a diagnostic role, especially in distinguishing frankly benign from overtly malignant forms.

Cyclin-D1-gene amplification and expression in breast carcinoma: relation with clinicopathologic characteristics and with retinoblastoma gene product, p53 and p21WAF1 immunohistochemical expression.

Cyclin D1 is a major positive regulator of the G1 restriction point promoting inactivation of the retinoblastoma protein (RB). The cyclin D1 gene is rearranged, amplified and/or over-expressed in several human neoplasms. In the present series of 64 human breast carcinomas, cyclin D1 amplification (4- to 8-fold) was seen in 24% of cases, and cyclin-D1 immunohistochemical over-expression was seen in 50% of cases. Amplification and over-expression were statistically associated; however, divergent result were seen in 30% of cases. Some of these discrepancies may reflect the fact that cyclin-D1 expression may be due to mechanisms other than gene amplification. Cyclin-D1 over-expression, but not cyclin-D1 amplification, was associated with positive oestrogen-receptor immunoreactivity. Cyclin-D1 amplification was associated with high RB expression, and 4 cases (7%) with absent RB immunoreactivity showed no cyclin-D1 amplification nor expression. Our data support the hypothesis that cyclin-D1 amplification may be associated with enhanced gene transcription and with high RB expression, that high ER expression may cooperate in maintaining high levels of cyclin-D1 protein, and that loss of RB function, as assessed by the lack of RB immunoreactivity, may be related to normal cyclin-D1 gene copy number and low cyclin-D1 expression.

Clinical significance of cyclin D1 expression in patients with node-positive breast carcinoma treated with adjuvant therapy.

BACKGROUND: Experimental and clinical studies suggest that cyclin D1 is involved in transformation and tumour progression. However, there is little and contradictory data on the clinical significance of cyclin D1 in human invasive breast carcinoma. PATIENTS AND METHODS: We investigated whether the determination of cyclin D1 has prognostic value in a series of 180 patients with node-positive breast carcinoma and treated with adjuvant therapy with a median follow-up exceeding 6 years. We assessed cyclin D1 expression using the CDS-6 monoclonal antibody and a highly sensitive immunohistochemical technique. RESULTS: We found that most of the evaluable tumours (117 of 167; 70.1%) presented nuclear cyclin D1 staining and that its expression was significantly associated with both the hormone receptors (P = 0.009 and P = 0.005 for ER and PgR, respectively). Furthermore, 29 (17%) of 167 tumours had a weak (15 cases) or strong (9 cases) cytoplasmic cyclin D1 staining. In a subgroup of cases we also studied the amplification of the cyclin D1 gene and a moderate agreement between cyclin D1 nuclear overexpression assessed immunohistochemically and the gene amplification was found. In univariate analysis, cyclin D1 nuclear positivity was significantly associated with improved 6-year relapse-free survival (RFS) (P = 0.004), but not with overall survival (OS) (P = 0.12). The results of the Cox multivariate analysis (final model) indicate that cyclin D1 expression (P = 0.0049) as well as the number of involved nodes (P < 0.001) and tumour size (P = 0.036) are significant prognostic indicators for RFS. Only the number of involved nodes retained significance (P < 0.001) for OS in our series. The joint assessment of the variables considered in the final model of the multivariate analyses had a moderate prognostic capability as determined using the Harrell c statistic (c = 0.66 and 0.64 for RFS and OS, respectively). CONCLUSIONS: The patients with node-positive breast cancer who have a higher likelihood of gaining benefit from adjuvant therapy are those with tumours with cyclin D1 nuclear expression, small size and less than 3 metastatic nodes. Further studies are needed to verify the prognostic value of cyclin D1 in relation to different adjuvant treatments and to deepen the biological pathways that regulate its activation/ suppression in human breast carcinoma.

[Audit of histopathological activities in the laboratories of 7 general hospitals. 1. Types of resources and quantitative aspects of the workload]. / Audit dell'attività istopatologica nei servizi di sette ospedali generali. 1. Tipi di risorse e aspetti quantitativi delle prestazioni.

Following a major reorganization of the National Health Service, competition for resources will depend to a great extent on the quality of the service offered, and audit will be of increasing importance. The present investigation deals with the quantitative aspects of the histopathology workload. A retrospective analysis of the biopsy handling of 1,000 consecutive cases was conducted in each histopathology lab of seven general hospitals. The data collected included information on specimen type, diagnosis, block/section/stain details for each specimen, as well as information about human resources, total workload and technical equipment. Among the seven labs, the histopathology workload ranged between 6,600 and 15,600 cases/year. The workload per person/year ranged between 1,400 and 2,600 (mean 1,900) for pathologists, between 800 and 3,000 (mean 1,500) for technicians, and between 2,200 and 8,300 (mean 4,400) for secretarial staff. The prevalence of biopsy fragments (endoscopic, endometrial curettage , etc.), "small" surgery (skin, appendix, gallbladder, etc.) and surgery was, respectively, between 42 and 50%, 34 and 47%, 9 and 21%. In all labs but one, 80% of cases were within 1-3 blocks; the seven labs had a mean of blocks/case ranging from 1.8 to 4.0 (total mean = 2.8). Differences in performing special stains were astonishing: one lab performed special stains in about 40%, and another in only 0.6% of cases (mean of the seven labs = 15%). Finally, the labs performed immunohistochemical stains within a range of 2.7-8% of cases (mean of the seven labs = 4.6%). The data we have collected provided baseline information on the laboratory running, and this is likely to be an integral part of laboratory administration in the near future.

Lipomas of the esophagus: a new case.

We present a new case of esophageal lipoma. The rarity of this tumor and its submucosal origin can create problems for the correct diagnosis. Symptoms, when present, are of mechanical origin (obstruction) with dysphagia. Treatment is endoscopic or surgical removal.

Relationship between cathepsin D and other pathological and biological parameters in 1752 patients with primary breast cancer.

The relationship between cathepsin D and other pathological or biological prognostic parameters has not yet been defined through systematic studies in breast cancer. The aim of the present investigation was to define the relationship between cathepsin D and nodal status, tumour size, steroid receptors and tumour grade in a wide patient series. Cytosol cathepsin D was assayed with an immunoradiometric assay in tumour samples from 1752 patients. A statistically significant, but not biologically meaningful association was found between cathepsin D and both tumour size and grade. Cathepsin D was significantly higher in node-positive than in node-negative tumours. However, cathepsin D is not of great use in order to predict the risk of axillary metastases in individual patients, due to overlapping of cathepsin D values between node-positive and node-negative cases. A significant, direct association was found between cathepsin D and both oestrogen receptor and progesterone receptor cytosol levels. Nevertheless, preliminary data indicate that cathepsin D and steroid receptors provide independent prognostic information.

Accuracy of bronchial aspiration cytology in typing operable (stage I-II) pulmonary carcinomas.

The accuracy of bronchial aspiration cytology in typing resectable (stage I-II) lung cancer has been investigated in 100 cases, comparing preoperative cytologic features with pulmonary tumor histology seen at surgery. The accuracy has been 100% for small-cell carcinoma (two cases), 98.8% for squamous-cell carcinoma (86 cases), and 91.6% for adenocarcinoma (12 cases). The overall accuracy rate has been 98%. No case of undifferentiated large-cell carcinoma has been identified. It is suggested that the high accuracy in cytologic typing of operable lung cancer is basically related to adequate preservation of differentiation features, thus allowing for correct identification of most non-small-cell carcinoma. Moreover, the absence in this study of any large-cell carcinoma, compared with its frequency in advanced stage series, would indicate that such a histotype reflects excessive dedifferentiation of an original squamous or glandular form.

Cytological typing of primary lung cancer: study of 100 cases with autopsy confirmation.

In order to assess the accuracy of bronchial aspiration cytology in typing lung cancer, tissue sections from 100 autopsy cases of lung cancer were compared with the cytology features observed in the same patients prior to death. There was 100% accuracy in the cytology of small-cell carcinoma; 90% in squamous-cell carcinoma; 70% in adenocarcinoma; and 50% in undifferentiated large-cell carcinoma. The observed discrepancies probably reflect intrinsic tumor properties rather than problems attributable to either the bronchial aspiration method or cytology interpretation, especially in cases involving advanced lung carcinoma. Because the highest accuracy rate was in detecting small-cell carcinoma, it is recommended that only the distinction between small-cell and non-small-cell forms be made on cytologic grounds and that further categorizations only be rendered in cases with unquestionable cytomorphological features.

Elastofibroma: CT and MR findings.

Two cases of elastofibroma of the chest wall are reported. Each patient was investigated with CT and, in 1 case, MR imaging was also performed. Elastofibroma appears like a mass in the subscapular region and often presents problems of differential diagnosis. The contribution of MR imaging is reported.

Hepatic and subcutaneous abscesses due to aspergillosis. Initial diagnosis of a case by intraoperative fine needle aspiration cytology.

Laparotomy of a 64-year-old woman with bleeding caused by a perforated anastomotic peptic gastric ulcer disclosed a mycotic abscess in the lower surface of the left hepatic lobe covering the ulcer. Fine needle aspiration (FNA) of the abscess produced a clear milky liquid that was positive for Aspergillus fungi by cytology and subsequent microbiologic and cultural studies. Postoperative diffusion of the aspergillosis into the subcutaneous tissue was similarly confirmed by cytology and Sabouraud solid medium culture. This appears to be the first case of disseminated aspergillosis diagnosed by intraoperative FNA cytology; it confirms that FNA cytology is a rapid, sensitive and important tool for the diagnosis of mycotic infections.

CT and MR imaging of melanocytic schwannomas; report of three cases.

Melanocytic Schwannomas are rare tumours which can arise in soft tissues, spinal nerve roots and in the central nervous system. The literature suggests that they have a malignant behaviour with local recurrence after surgery. We present three patients with this lesion, two in the thoracic spine and one in the head of the pancreas. The clinical outcome in these cases has been disappointing, since two patients died from complications due to local spreading of the tumour in spite of surgery and radiotherapy. CT could not distinguish these tumours from other neurogenic neoplasms. When these tumours occur near the vertebral body, CT could only detect early bone erosion. MR findings show promising features, with a high signal on T1-weighted images, due to melanin. However, more experience is necessary to establish the possible specific features of melanocytic Schwannomas in MRI.

Localization of hepatitis C virus (HCV) antigen by immunohistochemistry on fixed-embedded liver tissue.

An immunohistochemical method of staining HCV-related antigens in fixed-embedded liver biopsies is described. Two primary antisera were used: 1) a high titre anti-HCV human IgG separated from an anti-HCV positive serum; and 2) rabbit anti-HCV antibodies. In our experience this method proves to be reproducible and shows a good correlation with serologic results.