The Paradox Effect of Calcification in Carotid Atherosclerosis: Microcalcification is Correlated with Plaque Instability.

BACKGROUND: this study aims to investigate the possible association among the histopathologic features of carotid plaque instability, the presence of micro- or macrocalcifications, the expression of in situ inflammatory biomarkers, and the occurrence of the major risk factors in this process in a large series of carotid plaques. METHODS: a total of 687 carotid plaques from symptomatic and asymptomatic patients were collected. Histological evaluation was performed to classify the calcium deposits in micro or macrocalcifications according to their morphological features (location and size). Immunohistochemistry was performed to study the expression of the main inflammatory biomarkers. RESULTS: results here reported demonstrated that calcifications are very frequent in carotid plaques, with a significant difference between the presence of micro- and macrocalcifications. Specifically, microcalcifications were significantly associated to high inflamed unstable plaques. Paradoxically, macrocalcifications seem to stabilize the plaque and are associated to a M2 macrophage polarization instead. DISCUSSION: the characterization of mechanisms involved in the formation of carotid calcifications can lay the foundation for developing new strategies for the management of patients affected by carotid atherosclerosis. Data of this study could provide key elements for an exhaustive evaluation of carotid plaque calcifications allowing to establish the risk of associated clinical events.

Plaque calcification is driven by different mechanisms of mineralization associated with specific cardiovascular risk factors.

BACKGROUND AND AIMS: The aim of this study was to investigate possible associations among markers of mineralization, plaque instability and the main risk factors of atherosclerosis. METHODS AND RESULTS: A Tissue MicroArray containing 52 samples of calcified carotid plaques from 52 symptomatic and asymptomatic patients were built. TMA serial sections were used to study the expression of inflammatory and mineralization markers (BMP-2, BMP-4, VDR, RANKL, Osteopontin, Sclerostin, ß-catenin and calmodulin) by immunohistochemistry. Our data clearly demonstrated the expression of mineralization markers in atheromatic plaques. Indeed, with the exception of RANKL, all investigated markers were expressed in at least 60% of cases. Specifically, multivariate analysis displayed significant associations between both the expression of BMP-2 and the presence of unstable plaques as well as between the expression of ß-catenin and the presence of stable plaques. We also found a significant inverse association between both a) the presence of hypertension and VDR and b) smoking habits and calmodulin expression. Finally, we noted a higher density of RANKL positive cells in plaques from diabetic patients as compared to non-diabetic ones and a significant positive association between hypertriglyceridemia and BMP-4 expression. CONCLUSION: Our results support the hypothesis that the process of atherosclerotic plaque calcification presents a number of similarities with the physiological processes that occur in bone, involving both osteoblasts- and osteoclasts-like arterial cells. Finally, the present study suggests that risk factors, such as hypertension, cigarette smoke and diabetes, can cause the destabilization of the atheromatic plaque acting on calcification process as well as inflammation.

Prostate cancer and inflammation: A new molecular imaging challenge in the era of personalized medicine.

The relationship between cancer and inflammation is one of the most important fields for both clinical and translational research. Despite numerous studies reported interesting and solid data about the prognostic value of the presence of inflammatory infiltrate in cancers, the biological role of inflammation in prostate cancer development is not yet fully clarified. The characterization of molecular pathways that connect altered inflammatory response and prostate cancer progression can provide the scientific rationale for the identification of new prognostic and predictive biomarkers. Specifically, the detection of infiltrating immune cells or related-cytokines by histology and/or by molecular imaging techniques could profoundly change the management of prostate cancer patients. In this context, the anatomic pathology and imaging diagnostic teamwork can provide a valuable support for the validation of new targets for diagnosis and therapy of prostate cancer lesions associated to the inflammatory infiltrate. The aim of this review is to summarize the current literature about the role of molecular imaging technique and anatomic pathology in the study of the mutual interaction occurring between prostate cancer and inflammation. Specifically, we reported the more recent advances in molecular imaging and histological methods for the early detection of prostate lesions associated to the inflammatory infiltrate.

Ovarian Clear Cell Carcinoma: From Morphology to Molecular Biology.

Ovarian clear cell carcinoma (oCCC) is a distinctive subtype of ovarian carcinoma, with peculiar genetic and environmental risk factors, precursor lesions, molecular events during oncogenesis, patterns of spread, and response to treatment. Because of low response to chemotherapy and poor prognosis in advanced stages, there is growing interest in investigating the molecular pathways involved in oCCC development, in order to individualize novel/molecular targeted therapies. Until now, the main molecular genetic changes associated with oCCC remain to be identified, and, although several molecular changes have been reported in clear cell tumors, most studies have analyzed a limited number of cases; therefore, the true prevalence of those changes is not known. The present review will present the clinicopathologic features of oCCC, from morphology to molecular biology, discussing the diagnostic and treatment challenges of this intriguing ovarian carcinoma.

Cytologic Diagnosis of Oncocytic Neoplasms of the Thyroid Gland: The Importance of the Clinical Scenario.

It is a diagnostic challenge to differentiate benign and malignant thyroid neoplasms made up of Hürthle (or oncocytic) cells on cytologic material. They are large, polygonal cells with marked eosinophilic, granular cytoplasm reflective of overly abundant mitochondria. These cells commonly occur in nodular goiters and dominant adenomatous or hyperplastic nodules though they may also be the predominant component of neoplastic lesions. There are significant controversies concerning the optimal management of patients with oncocytic cell carcinoma. This review provides an overview of the most significant studies addressing the distinction between benign and malignant Hürthle cell lesions on cytology and histology.

Small Bowel Carcinomas in Coeliac or Crohn's Disease: Clinico-pathological, Molecular, and Prognostic Features. A Study From the Small Bowel Cancer Italian Consortium.

BACKGROUND AND AIMS: An increased risk of small bowel carcinoma [SBC] has been reported in coeliac disease [CD] and Crohn's disease [CrD]. We explored clinico-pathological, molecular, and prognostic features of CD-associated SBC [CD-SBC] and CrD-associated SBC [CrD-SBC] in comparison with sporadic SBC [spo-SBC]. METHODS: A total of 76 patients undergoing surgical resection for non-familial SBC [26 CD-SBC, 25 CrD-SBC, 25 spo-SBC] were retrospectively enrolled to investigate patients' survival and histological and molecular features including microsatellite instability [MSI] and KRAS/NRAS, BRAF, PIK3CA, TP53, HER2 gene alterations. RESULTS: CD-SBC showed a significantly better sex-, age-, and stage-adjusted overall and cancer-specific survival than CrD-SBC, whereas no significant difference was found between spo-SBC and either CD-SBC or CrD-SBC. CD-SBC exhibited a significantly higher rate of MSI and median tumour-infiltrating lymphocytes [TIL] than CrD-SBC and spo-SBC. Among the whole SBC series, both MSI─which was the result of MLH1 promoter methylation in all but one cases─and high TIL density were associated with improved survival at univariable and stage-inclusive multivariable analysis. However, only TILs retained prognostic power when clinical subgroups were added to the multivariable model. KRAS mutation and HER2 amplification were detected in 30% and 7% of cases, respectively, without prognostic implications. CONCLUSIONS: In comparison with CrD-SBC, CD-SBC patients harbour MSI and high TILs more frequently and show better outcome. This seems mainly due to their higher TIL density, which at multivariable analysis showed an independent prognostic value. MSI/TIL status, KRAS mutations and HER2 amplification might help in stratifying patients for targeted anti-cancer therapy.

Primitive "Spindle Cell Variant" (Sarcomatoid Variant) Diffuse Large B-Cell Lymphoma of the Uterine Cervix: Description and Outcome of a Rare Case.

A very rare case of primary diffuse large B-cell lymphoma of the uterine cervix characterized by "spindle cell variant" morphology ("sarcomatoid subtype") is described along with a discussion of the challenging diagnosis due to its rarity and presenting clinical and pathological features.

Well-differentiated thyroid cancer with a minor poorly differentiated component: clonal heterogeneity through the prognostic role of CXCR4 and BRAF analysis.

Well-differentiated carcinoma (WDC) accounts for up to 90% of all thyroid cancers. The presence of a minor poorly differentiated (PD) component (mainly insular pattern) might represent an additional critical parameter for patients' prognosis and outcome. The role of both CXCR4 (a chemokine inducing cytoskeletal rearrangement and cell adhesion) and BRAF mutation have been studied in WDC (mainly papillary thyroid cancer and its variants), highlighting their critical role in tumor progression, local infiltration, and metastases. We discussed the clonal heterogeneity through the prognostic role of CXCR4 and BRAF mutation in WDC with a minor PD/insular component. Of our 16 WDC cases with a PD/insular component, up to 40% underwent surgery. The cases were subclassified according to the PD percentage as (1) <20% PD and (2) 20% to 40% PD, and were studied for CXCR4 expression and BRAF mutation. CXCR4 and molecular testing were distinctly performed on both components of each lesion. The majority of the cases (69%) showed an extrathyroid and metastatic dissemination. Regardless of the 2 categories, we had 8/16 (50%) patients with disease-free status. CXCR4 was negative in all 16 cases, whereas 3 of them (19%) had a mutated BRAF only in the WDC component of the lesion. WDCs with a minor PD pattern, even when <20%, showed more aggressive features than pure WDCs and should be entirely considered as PD carcinoma. The absence of CXCR4 expression and BRAF mutation in cancers with a minor PD component underlined different pathogenic and metastatic processes in comparison with WDCs.

Primary pulmonary hodgkin lymphoma simulating a mediastinal tumour: an uncommon occurrence.

The case of a patient with primary pulmonary Hodgkin Lymphoma simulating a mediastinal tumour is reported for its rarity and the diagnostic concerns encountered by us.

Diagnostic and prognostic role of HBME-1, galectin-3, and ß-catenin in poorly differentiated and anaplastic thyroid carcinomas.

AIM: Thyroid cancer represents the first endocrine malignant neoplasm, accounting for 1% of human malignancy. The majority of which are well-differentiated cancer representing up to 90% of thyroid cancer and pursuing a favorable clinical course. The groups of poorly differentiated thyroid cancer (PDC) and anaplastic thyroid cancer (ATC) have a poor outcome and need a strict clinical surveillance. MATERIALS AND METHODS: Thirty-four cases including 23 PDC/insular cancer and 9 ATC were examined for the expression of an immunohistochemical panel made up by HBME-1, galectin-3, and ß-catenin and correlated either with histologic prognostic parameters or the overall surveillance. RESULTS: HBME-1 and galectin-3 were expressed in 100% of the PDC/insular cases and in none of the ATC cases. The data for ß-catenin pointed out an 80% expression (12/15) in the PDCs and only a focal and nonspecific positivity in the ATCs. A ß-catenin-positive expression was found in all patients with a worse outcome/death and in the presence of vascular invasion and metastatic disease. All 3 PDC patients with ß-catenin negativity are alive, whereas only 41% (5/12) are alive in the ß-catenin-positive group. CONCLUSIONS: Our data set up the idea that PDC represents an intermediate step in the biological process of dedifferentiation of thyroid tumors toward ATC. This shift is underlined by the ß-catenin expression, which seems to be related to a worse prognostic behavior. HBME-1 and galectin-3 show a similar pattern in PDC compared with well-differentiated carcinoma, whereas they are not expressed, as well as ß-catenin, in anaplastic carcinomas.

Application of liquid-based cytology to fine-needle aspiration biopsies of the thyroid gland.

Fine-needle aspiration biopsy is regarded as an important tool for diagnosing thyroid lesions because of its simplicity, safety, and cost-effectiveness. Its role in correctly characterizing the group of indeterminate lesions or follicular-patterned neoplasms (FN) might be more decisive. Liquid-based cytology (LBC) is a technique based on the use of a semi-automated device that has gained popularity as a method of collecting and processing both gynecologic and non-gynecologic cytologic specimens. It achieves a diagnostic sensitivity as accurate as conventional preparations especially for its excellent cell preservation and for the lack of background which decrease the amount of inadequate diagnoses. Moreover, the cellular material which has been stored in the preservative solution could be effectively used for the application of immunocytochemical and molecular techniques especially for the Follicular proliferations. In many cases the cytologic features are similar in both methods but the colloid film and the lymphocytic component are more easily evaluated on direct smears whereas nuclear details and colloid globules are better evaluated in LBC slides. The LBC-processed biopsies represent a valid alternative to conventional cytology. The possibility of applying special techniques enhance the efficacy of the cytological diagnosis of thyroid lesions.

Mucinous adenocarcinoma of the urinary bladder.

BACKGROUND: Mucinous adenocarcinoma of the urinary bladder is a rare primary urologic disease, poorly responsive to radiation or chemotherapy as first-line treatment. CASE REPORT: After trans-urethral resection of the bladder, a 62-year-old woman was diagnosed with mucinous adenocarcinoma of the urinary bladder. An upper gastro-intestinal endoscopy and a colonoscopy excluded any primary site of origin from those gastro-intestinal tracts. After whole-body CT staging scans, an anterior pelvectomy was performed, confirming a mucinous adenocarcinoma of the bladder, with no extra-vesical spreading. Some onco markers were sampled before surgery, and Ca 19-9 showed very high values, with a decreasing trend after pelvectomy. Six month after surgery, bilateral inguinal lymph node dissection was performed because of bilateral palpable masses - histologic examination showed a single metastatic node. The patient also received external radiotherapy of the inguinal area. Twenty-eight months after pelvectomy, the patient appears healthy. CONCLUSIONS: Early radical surgery with or without adjuvant radio-chemo-therapy appears to be the best option for mucinous adenocarcinoma of the bladder, and a good outcome is likely to be related with a confined disease and small tumor size. In addition, Ca 19-9 sampling proves to be useful in tumors that produce markers.

Correlation between pathological data and the RNA expression of p53 or p53-targeted genes in primary invasive ductal breast carcinomas: a preliminary study.

The protein expression of the growth suppressive p53 transcription factor and its inhibitor human double minute 2 (Hdm2) is altered in ductal breast carcinomas (DBC). However, the assessment of p53 and/or Hdm2 protein levels in DBC tissues was found to have a questionable prognostic significance. We evaluated the RNA expression of p53, hdm2, and the p53-targeted p21waf-1 and thrombospondin (tsp)-1 by primary DBC tissues, then correlated the RNA levels with patient clinicopathological data. The mean RNA expression level of p53 and that of hdm2 were elevated in large-sized, poorly differentiated, node-positive DBC, while a high p21waf-1 or tsp-1 mean expression level comprised small-sized, low-grade, node-negative tumors. Further analyses found that the correlation between the RNA expression of p53 and that of its targeted genes was reduced as tumor aggressiveness increased. However, for all the examined genes, association of the intensity of RNA expression with the pathological data was not statistically significant (p>0.05). Altogether, our preliminary RNA data confirm the results from previous protein studies, indicating that despite p53 expression and activity show a trend to vary in association with DBC clinical features, neither p53 nor its transcriptional targets can accurately monitor the behaviour of invasive DBC.

Laparoscopic resection of sporadic synchronous gastric and jejunal gastrointestinal stromal tumors: report of a case.

Multicentricity of gastrointestinal stromal tumors (GISTs) has been described only in patients with neurofibromatosis type 1 (NF1) or within the small intestine, and different pathogenetic mechanisms are involved. We report a case of synchronous sporadic gastric and jejunal GISTs, which were resected laparoscopically in a 67-year-old man. Immunohistochemical analysis revealed that both lesions were KIT (CD117)-positive, but that the gastric lesion was CD34-positive, whereas the jejunal one was Vimentin-, S-100-, and SMA-positive. Molecular analysis of mutations in KIT exons 9, 11, 13, and 17, and in PDGFRA exons 12 and 18 revealed the presence of a gastric sporadic GIST with a KIT mutation of the exon 11 and a jejunal sporadic GIST without KIT or PDGFRA mutations. To our knowledge, this is the first report of laparoscopically resected synchronous sporadic gastric and jejunal GISTs.

Relevance of immunocytochemistry on thin-layer cytology in thyroid lesions suspicious for medullary carcinoma: a case-control study.

BACKGROUND: Fine needle aspiration cytology represents the most important tool in the diagnosis of thyroid nodules, mostly in discriminating malignant from benign lesions. The diagnosis of medullary thyroid carcinoma (MTC) may present some problems related to its deceptive morphologic picture. This diagnosis may be supported by immunocytochemistry (ICC), which may be difficult to carry out on the conventional smears. DESIGN: The diagnostic efficacy of ICC for the diagnosis of MTC with respect to other thyroid neoplasms on slides processed by thin-layer cytology (TLC) is evaluated. PATIENTS: In the period between January 2002 and December 2005, 8,200 FNAB were processed. ICC on TLC slides was required in 33 cases. Conventional smears were fixed in ethanol, whereas TLC slides were processed with the Thin Prep 2000 method. All slides were then stained with Papanicolaou. In all cases where MTC was morphologically suspected, ICC for calcitonin, monoclonal carcinoembryonic antigen, and thyroglobulin was carried out only on TLC slides. RESULTS: Thirty-three thyroid cytologic cases had ICC on the TLC slides, including 22 follicular proliferations and 11 malignant lesions. The application of ICC on TLC was conclusive in 32 cases and inconclusive in 1 case. Twenty cases underwent surgery. No false-positive and false-negative cases were found. Sensitivity and specificity were 100%, and the overall diagnostic accuracy was 100%. CONCLUSIONS: ICC can be successfully applied on TLC slides. The combined results of morphology and a small immunopanel including thyroglobulin, calcitonin, and carcinoembryonic antigen yields a 100% diagnostic efficacy for MTC. CONDENSED ABSTRACT: Fine needle aspiration cytology is an excellent technique for diagnosing malignant neoplasms of the thyroid, especially those derived from the follicular cells. A correct preoperative diagnosis of C-cell-derived tumors (MTC), which is essential for both the surgical approach to the primary tumor and the management of the patient, should rely not only on the morphologic picture but also on the immunocytochemical yielding using an immunopanel, which is particularly satisfactory on the TLC slides.

Solitary fibrous tumour of thyroid: report of two cases with immunohistochemical features and literature review.

Solitary fibrous tumour (SFT) is a rare tumour principally found in adults in the pleural cavity. Extrapleural occurrences are rare. Two cases of SFT of the thyroid gland are described in this paper showing their distinctive microscopical architecture, namely "patternless growth pattern". It is characterized by a bland spindle-cell proliferation alternating hyper- and hypo-cellular areas, keloid-like hyalinization and a focal hemangiopericytoma-like vascular pattern. Tumour cells revealed a diffuse strong positivity for CD34, CD99, bcl-2 and Vimentin, but negativity for Desmin, EMA, AE1/AE3, SMA, S-100 and CD31 antibodies. The differential diagnosis of thyroid SFT includes different types of spindle cell proliferation, benign and malignant mesenchymal tumours, medullary thyroid carcinoma, fasciitis-like papillary carcinoma, and undifferentiated (anaplastic) carcinoma. However, the morphologic and immunohistochemical findings of SFT are so characteristic that this diagnosis seldom represent a difficulty.

Apoptotic cells are present in ischemic zones of deep partial-thickness burns.

Deep partial-thickness burns exhibit ambiguous behavior, either spontaneously healing or evolving into full-thickness burns. The aim of this study was to investigate these lesions for the presence of apoptotic cells and to compare their rate with that of superficial and full-thickness burns. We used colocalization of DNA fragments (ie, terminal deoxynucleotidyl transferase Biotin-dUTP nick end labeling) and Fas ligand CD95 antibodies to calculate the apoptotic rate of superficial, deep partial-thickness and full-thickness burns in 45 patients after the thermal injury. Biopsies were collected mainly during the acute postburn phase (first week of hospitalization). Deep partial-thickness burns presented apoptotic cells, both in the dermis and in cutaneous adnexa, and showed a higher apoptotic rate than superficial and full-thickness burns (44.5% in deep partial thickness, interquartile range 6.3-90.5%; 5.6% in superficial partial thickness, interquartile range: 0-13%; 0% for full-thickness burn; P = .000243). A significant greater apoptotic rate was present in cells of deep partial-thickness burns when compared with superficial and full thickness. These data would suggest that deep burns sustain an ischemic damage that forces cells to undergo apoptosis and could represent the biologic basis for their clinical evolution into full-thickness burns. Further correlation studies are now required to confirm this hypothesis.

Tissue microarray analysis of FAS, Bcl-2, Bcl-x, ER, PgR, Hsp60, p53 and Her2-neu in breast carcinoma.

BACKGROUND: The aim of this study was to detect immunohistochemical markers in breast carcinoma by means of tissue microarray analysis (TMA) and to associate their expressions with clinicopathological features and prognosis. Fatty acid synthase, bcl-2, bcl-x, p53, estrogen and progesterone receptors, heat shock protein 60 and Her2-neu (c-erbB-2) were evaluated in a group of 149 breast carcinoma patients with a 5-year follow-up period. MATERIALS AND METHODS: TMA blocks were made by using duplicate 0.6-mm diameter tissue cores from each paraffin block. RESULTS: Statistical analysis revealed that tumor stage (p=0.003) and node status (p=0.001) were the only two prognostic markers of disease-free survival. Moreover, FAS and bcl-x showed an independent effect on recurrence (p=0.005). The node status was the only marker of overall survival (p=0.05). CONCLUSION: Our data confirmed recent reports associating the stage of disease, FAS and Bcl-x expressions with recurrence and outcome. These data demonstrated that TMA is an effective substitute for conventional histochemical-immunohistochemical techniques.