Detection of an EGFR mutation in cytological specimens of lung adenocarcinoma.

OBJECTIVE: Epidermal growth factor receptors (EGFR) mutation status is crucial for the prediction of a tumour response to treatment with EGFR tyrosine kinase (EGFR-TK) inhibitors. The aim of the study was to establish a protocol for the detection of EGFR-activating somatic mutations on cytological samples collected using a standard bronchoscopy procedure and to determine the frequency of EGFR mutations among pre-selected Croatian patients with non-small cell lung cancer (NSCLC) of an adenocarcinoma histological subtype. METHODS: A total of 177 cytological samples were collected from the patients diagnosed with NSCLC. DNA was isolated from the cytological material recovered from the fixed and stained slides. EGFR mutations were analysed using the polymerase chain reaction (PCR)- mediated Sanger sequencing method. RESULTS: Out of 177 collected samples, EGFR mutation analyses were successfully performed on 167 samples (94.4%); 77 (46.1%) of these were from male and 90 (53.9%) from female patients. EGFR mutations/deletions were found in 33 (19.8%) of the tested patients; exon 19 deletions in 17 (10.2%) and point mutations of exon 21 in 16 (9.6%) patients. CONCLUSION: The PCR-mediated Sanger sequencing method was found to be reproducible and reliable. Cytological samples can be used successfully to determine the EGFR mutation status in NSCLC patients providing information for targeted therapy at an early stage of the disease.

EGFR expression is linked to osteopontin and Nf-κB signaling in clear cell renal cell carcinoma.

AIM AND BACKGROUND: Epidermal growth factor receptor (EGFR) is a receptor tyrosine kinase involved in many important aspects of cell biology that are related to tumorigenesis. There are opposite evidences of the role of EGFR in renal cancer and the outcome of EGFR-targeted therapies, suggesting the complexity of EGFR signaling pathways. In vitro, osteopontin (OPN) and nuclear factor kappa B (NF-κB) are thought to be involved in specific ligand-independent EGFR activation that could have a role in resistance to EGFR mAb therapy. Aim of this study was to analyze the relationship between EGFR and OPN at the protein and mRNA level, as well as their relation to NF-κB in clear cell renal cell carcinoma (CCRCC). MATERIALS AND METHODS: Expression of EGFR, OPN, and p65 NF-κB protein was analyzed using immunohistochemistry and compared mutually in 88 CCRCC samples. Expression of EGFR and OPN mRNAs was analyzed using quantitative Real-time PCR in 22 CCRCC samples and compared mutually and with NF-κB protein expression. RESULTS: Epidermal growth factor receptor mRNA level was higher in CCRCC samples in comparison with normal renal tissue (p = 0.012) and was associated with high OPN mRNA level, and with NF-κB activation (p < 0.001 and p = 0.045, respectively). Immunohistochemical staining showed the inverse association; high EGFR protein expression was related with low OPN and NF-κB protein expression (p < 0.001 and p = 0.047, respectively). CONCLUSION: Epidermal growth factor receptor gene is upregulated in CRCC and associated with OPN gene expression and NF-kB signaling. The inverse relation between OPN and EGFR at the protein level could probably reflect dynamic changes that EGFR undergoes following activation.

Apoptotic cell death and rupture of abdominal aortic aneurysm.

The abdominal aortic aneurysm (AAA) development and expansion is characterized by an extracellular matrix degradation and widespread inflammation. In contrast, the processes that characterize the AAA rupture are not completely understood. The results obtained in animal and clinical studies have shown the importance of inflammation, proteolysis, and antioxidant mechanisms in the aortic degeneration and formation of AAA. We hypothesize that the rupture of the AAA could have a similar pathway like an atherosclerotic plaque rupture, and in both the cases the apoptotic cell death of smooth muscle cells could play a significant role. If the apoptotic cell death significantly contributes to the expansion and rupture of aneurysm, the hypothesis is that aggressive medical antiapoptotic treatment with high doses of appropriate drugs could decrease the apoptotic index of smooth muscle cells, reduce the aneurysm expansion and prevent rupture.

Primary leiomyosarcoma of the tibia: five years without recidivism

Primary leiomyosarcoma of the bone is a rare malignancy. Clinical follow-up suggests that primary osseous leiomyosarcoma has an aggressive biologic behaviour with poor survival time. We report a case of primary leiomyosarcoma arising from the proximal metaphysis of the right tibia of a 55-year old woman with a long follow-up period, without recidivism. Primary leiomyosarcoma has to be considered as a differential diagnostic possibility in the case of bone tumours seen on routine initial plain radiographs as lytic lesions. If the tumour has been adequately excised at the time of diagnosis, as in the present case, with adjuvant therapy protocol, the long-term prognosis of such an aggressive tumour can be exceptionally good.

El leiomiosarcoma primario del hueso es un tumor maligno raro. El seguimiento clínico sugiera que el leiomiosarcoma óseo primario tiene un comportamiento biológico agresivo con escaso tiempo desobrevivencia. Reportamos un caso de leiomiosarcoma primario proveniente de la metáfisis proximal de la tibia derecha de una mujer de 55 años de edad con un largo período de seguimiento, sin recidivas. El leiomiosarcoma tiene que ser considerado una posibilidad de diagnóstico diferencial en el caso detumores óseos observados en radiografías simples iniciales de rutina como lesiones líticas. Si el tumorha sido debidamente extirpado en el momento del diagnóstico – como en el caso presente – con un protocolo de terapia adyuvante, la prognosis a largo plazo de un tumor tan agresivo puede ser excepcionalmente buena.

Primary leiomyosarcoma of the tibia--five years without recidivism.

Primary leiomyosarcoma of the bone is a rare malignancy. Clinical follow-up suggests that primary osseous leiomyosarcoma has an aggressive biologic behaviour with poor survival time. We report a case of primary leiomyosarcoma arising from the proximal metaphysis of the right tibia of a 55-year old woman with a long follow-up period, without recidivism. Primary leiomyosarcoma has to be considered as a differential diagnostic possibility in the case of bone tumours seen on routine initial plain radiographs as lytic lesions. If the tumour has been adequately excised at the time of diagnosis, as in the present case, with adjuvant therapy protocol, the long-term prognosis of such an aggressive tumour can be exceptionally good.

Macrophage level is not affected by monocyte chemotactic protein-1 in invasive ductal breast carcinoma.

PURPOSE AND METHOD: Monocyte chemotactic protein-1 (MCP-1) is a chemokine involved in the macrophage infiltration of tumor tissue. Tumor-associated macrophages (TAMs) are a population of mononuclear phagocytic cells that can have a complex function in tumor biology. The aim of this study was to determine the possible correlation between parenchymal MCP-1 expression and TAM level by immunohistochemical analysis of 97 invasive ductal breast carcinomas, not otherwise specified (NOS), and to investigate their relation with tumor size, histological grade, mitotic activity index (MAI) and lymph node status. Secondly, the MCP-1 mRNA was determined by reverse transcriptase-polymerase chain reaction (RT-PCR) in eight samples of normal breast tissue and 27 samples of invasive breast carcinomas and compared with TAMs. RESULTS: MCP-1 immunoreactivity was present in tumor cells (17/97), but also in TAMs, fibroblasts and endothelial cells. The statistical analysis did not show a significant correlation between MCP-1 expression in tumoral epithelium and tumor size, histological grade, MAI, lymph node status or TAMs. The results of RT-PCR showed that, in all cases of breast carcinomas (27/27) and the majority of normal breast tissues (7/8), the number of detected MCP-1 cDNA copies was above the detection limit. However, carcinomas showed higher levels of MCP-1 mRNA than normal breast tissue. Nevertheless, the statistical analysis did not find a significant correlation between MCP-1 expression and macrophage infiltrations. CONCLUSION: These results indicate that MCP-1 is probably not the only and/or crucial factor involved in macrophage attraction to tumor locus in breast carcinoma.

Primary non-Hodgkin lymphoma of the humerus following traumatic injury: case report.

A case of primary non-Hodgkin lymphoma of the right humerus which occurred in a 21-year-old male patient after an impact to the right shoulder in a car accident in July 1983 is described. Seventeen years after the injury, due to a civil lawsuit, the biopsy material was revised. Immunohistochemical analysis showed CD20 and CD79a positivity on large pleomorphic cells, while small reactive lymphocytes were CD3, Bcl-2 and CD20 positive. Molecular analysis carried out with PCR revealed a monoclonal B-lymphocyte population. The diagnosis of diffuse large peripheral B cell lymphoma of the bone was confirmed. The present case concurs with the literature on primary bone lymphoma, in which the diagnostic problem, trauma-related presentation and an excellent prognosis of malignant tumour are emphasized.

Cytomorphological variations, proliferation and angiogenesis in the prognosis of cutaneous melanoma.

Depth of invasion and stage of the disease are well known prognostic indicators in cutaneous melanoma (CM). However, the role of other parameters, such as the variations in cytomorphology of melanocyte tumours, mitotic activity and angiogenesis is still open to question. The aim of this study was to analyse proliferation by mitotic activity index (MAI) and immunostaining of proliferating cell nuclear antigen (PCNA), and the intensity of neovascularization (microvessel density; MVD) in CM clinical stage I in relation to epithelioid, spindle and nevoid cell type, histological type (superficial spreading melanoma and nodular melanoma), Clark's level and Breslow thickness. Finally, the role of all parameters in the prognosis of CM was evaluated. Statistical analysis demonstrated that cytological characteristics of CM correlate only with Clark's level, while histological types correlate with MAI, PCNA and MVD. MAI and PCNA also showed correlation between groups according to Clark's level and Breslow thickness. Finally, tumour cell PCNA was found to correlate with MVD. Survival of patients with CM correlated significantly with MAI. These results suggest that cytological variation, histological type, PCNA and MVD alone are not independent prognostic parameters, whereas MAI is a potentially important prognostic marker in CM.

Influence of tumor thickness and vascularity on survival in cutaneous melanoma.

Eighty-eight cases of cutaneous melanoma (CM) were analyzed in order to elucidate the relationship between thickness, angiogenesis, and prognosis. The thickness of the tumor was measured according to the Breslow method, and the microvessels were identified by an immunohistochemical study using anti-factor VIII monoclonal antibody on specimens from 40 patients with superficial spreading melanoma (SSM), and 48 with nodular type (NM). Microvessels were counted in the area of highest density. The overall survival and disease-free period were analyzed retrospectively. The proportion of patients with thicker CMs (> 1.5 mm) increased with age in both sexes. Mean vascular count was statistically significant different only between thinner and thicker tumors in the SSM group (P < 0.05). Prognosis was correlated with the thickness of CM (P = 0.0002), mean vascular count alone (P = 0.004), mean vascular count in association with CM thickness less than 1.5 mm (P = 0.0005), and with mean vascular count in NM (P = 0.02). These findings suggest that increasing microvessel density indicates a worsening prognosis.

[Intra-abdominelle Lymphangiome]. / Intra-abdominal lymphangioma - Case Report.

Intra-abdominal lymphangiomas are very rare. We report the unusual case of a 4-year-old boy, who presented with acute abdominal symptoms due to intracystic hemorrhage following a blunt abdominal trauma. After the diagnostic management, the radical resection was done with a good outcome.

Gastric epithelium proliferation in early Hp+ and Hp- gastritis: a flow cytometry study.

BACKGROUND: Helicobacter pylori increases cellular turn-over causing hyperproliferation and possible assumption of neoplastic characteristics by the gastric epithelial cells. To verify whether patients at risk of cancer can be identified at the very first stages of gastric disease, we studied a sample of patients affected by early Hp+ and Hp- gastritis by flow cytometry and assessed the methods commonly adopted to study gastric cell proliferation. METHODS: 48 fresh biopsies taken from the gastric antrum and body of 24 patients who had undergone endoscopy for dyspepsia, and 48 paraffin-embedded antrum and body biopsies taken from the files of our Department were studied by flow-cytometry. The following markers and parameters were considered: S-phase, proliferation index, PCNA and ploidy. RESULTS: No correlation was found between Hp+ or Hp- gastritis and gastric cell proliferation and no cases of aneuploidy were observed. Gastric proliferation was found to vary depending on the methods, markers and type of biopsy employed. Furthermore, proliferation expressed by PCNA was significantly different in antrum and body. CONCLUSIONS: The commonly studied proliferation markers do not allow the early detection of patients at risk of gastric cancer by flow cytometry. Proliferation differences between body and antrum must be taken into account in the investigation of gastric diseases.

Carl von Rokitansky and the Italian translation of the Handbuch der Pathologischen Anatomie: a linguistic and doctrinal enigma.

Carl von Rokitansky was the author of a treatise that came out between 1842 and 1846 with the title Handbuch der Pathologischen Anatomie. A historical milestone in pathological anatomy, Rokitansky's work represented the first attempt to systematically classify pathological specimens. Its publication inevitably made a great impact on Vienna, at that time the major European medical centre. The Italian translation of Rokitansky's masterpiece, Trattato Completo di Anatomia Patologica, published in Venice in 1852, was carried out by Ricchetti and Fano: the former a philologist and the latter a Triestine physician who, in 1873, had worked at Simon Pertot's side, the first prosector to be assumed in Trieste. From the start, the two translators not only made no secret of the linguistic obstacles they came up against, but also how unconvincing Rokitansky's doctrines were; a scepticism emerged from their words that inevitably contributed to the realization of a translation difficult to read. Undoubtedly, Rokitansky elaborated a theory of disease containing a certain degree of unclarity and in this respect it is interesting to emphasize that even the English translation, Manual of Pathologic Anatomy (1849-1854), demonstrated similar conceptual problems. A convinced supporter of gross pathology, Rokitansky put forth a theory of disease, the so-called Krasenlehre, resting upon humoral doctrines. This new knowledge inevitably exerted a great influence over Viennese, as well as German, medicine. Rokitansky's humoral pathology survived until the 1850s, when it was attacked by young Virchow, the future, universally recognized, father of cellular pathology, who definitively extinguished speculative humoral pathology.

Prognostic significance of DNA ploidy pattern and nucleolar organizer regions (AgNOR) in colorectal carcinoma.

AIM: To investigate the prognostic significance of DNA ploidy and silver stained nucleolar organizer regions (AgNOR), as well as their relation to the histological grade and Dukes' stages of colorectal carcinomas, and the relation of tumor cells proportion in the S-phase and Dukes' stage, histologic grade, DNA ploidy, or AgNOR count. METHODS: DNA flow cytometric analysis and AgNOR were performed on 94 surgically removed colorectal carcinomas. The mean AgNOR count was calculated in 200 tumor cells for each case. Survival rates and tests for significance were evaluated using the log-rank test and Cox regression model. RESULTS: There were no significant correlations between the ploidy pattern, histological grade, and Dukes' stage. Diploid tumors had a significantly lower AgNOR count (median 2.5, range 2.1-7.7) than aneuploid (median 6.2, range 2.0-7.9). Dukes' C stage tumors exhibited higher AgNOR count than Dukes' A or B stages. The proportion of tumor cells in S-phase did not correlate with any other parameter. Each of these parameters failed to show any correlation with survival. After dividing the tumors into those with high (>5) and low AgNOR count (<5), no correlation was found in the latter group between AgNOR and any other studied parameters, whereas in the group with high AgNOR count correlations to Dukes' stage, DNA ploidy, and histological grade were established.Conclusions. The difference in survival between well, moderately, and poorly differentiated tumors were significant in the group with high AgNOR counts. Dukes' C stage and aneuploid tumors had the worst prognosis.

Monoclonal antibodies against maternal major histocompatibility complex class I molecules induce rapid abortion in mice.

PROBLEM: The role of antibodies against fetal or maternal antigens in maintaining or losing pregnancy is not clear. METHOD OF STUDY: Term-pregnant mice were injected with monoclonal antibodies against only fetal or fetal and maternal major histocompatibility complex class I molecules. The development of pregnancy was then followed. RESULTS: Antibodies against maternal, but not fetal, major histocompatibility complex class I molecules induced abortion in mice. The abortion occurred 6-8 hr after the administration of autoreactive antibodies. The abortion could only be induced after the formation of placenta. Antibodies against tumor necrosis factor-alpha could not prevent or postpone the abortion. Extensive bleeding has been detected in the placenta of aborting mice 3 hr after the administration of the antibodies. CONCLUSIONS: This study indicates that autoreactive antibodies present risk for pregnancy and that the damage leading to abortion induced by such antibodies most likely occurs at the maternal side of placenta.

CD30-positive cutaneous anaplastic large cell lymphoma with ichthyosis acquisita.

Anaplastic large cell lymphoma (ALCL) is a high-grade non-Hodgkin's lymphoma recognized by the expression of the CD30 marker and by its morphology. We report an unusual case of ALCL in a 42-year-old woman. For ten years the patient only had pruritic erythematous skin lesions, then a plaque with nodules and ulcers on the right thigh and leg developed, followed by ichthyosis acquisita. The development of ALCL in women and the association of ichthyosis acquisita with this type of lymphoma are uncommon.