Expression profiling of extramedullary acute myeloid leukemia suggests involvement of epithelial-mesenchymal transition pathways.

Extramedullary (EM) colonization is a rare complication of acute myeloid leukemia (AML), occurring in about 10% of patients, but the processes underlying tissue invasion are not entirely characterized. Through the application of RNAseq technology, we examined the transcriptome profile of 13 AMLs, 9 of whom presented an EM localization. Our analysis revealed significant deregulation within the extracellular matrix (ECM)-receptor interaction and focal-adhesion pathways, specifically in the EM sites. The transcription factor TWIST1, which is known to impact on cancer invasion by dysregulating epithelial-mesenchymal-transition (EMT) processes, was significantly upregulated in EM-AML. To test the functional impact of TWIST1 overexpression, we treated OCI-AML3s with TWIST1-siRNA or metformin, a drug known to inhibit tumor progression in cancer models. After 48 h, we showed downregulation of TWIST1, and of the EMT-related genes FN1 and SNAI2. This was associated with significant impairment of migration and invasion processes by Boyden chamber assays. Our study shed light on the molecular mechanisms associated with EM tissue invasion in AML, and on the ability of metformin to interfere with key players of this process. TWIST1 may configure as candidate marker of EM-AML progression, and inhibition of EMT-pathways may represent an innovative therapeutic intervention to prevent or treat this complication.

The Efficacy and Safety of Lenalidomide plus Rituximab in an Orbital Relapse of Diffuse Large B-Cell Lymphoma.

A 74-year-old male with diffuse large B-cell lymphoma, with an Ann Arbor stage IV-A, was submitted to immune-chemotherapy in 2014, with complete remission of the disease. Two years later, he presented with a left eye swelling leading to exophthalmos and blurred vision. A core biopsy was performed and revealed a local relapse of the disease. He was considered unfit for intensive salvage chemotherapy and was treated with a combination of rituximab and lenalidomide. After six courses of rituximab plus lenalidomide, the patient showed complete remission and was submitted to maintenance therapy with lenalidomide. After 24 months since the start of lenalidomide monotherapy, we did not observe any progression. In this experience, rituximab plus lenalidomide, without radiotherapy, was a safe and effective therapeutic combination in an elderly patient with a localized relapse of DLBCL who was unfit to receive more aggressive therapies.

Carotid plaque instability is not related to quantity but to elemental composition of calcification.

BACKGROUND AND AIMS: Recent studies highlighted the role of calcification processes in the clinical progression of chronic cardiovascular diseases. In this study we investigated the relationship between the chemical composition of calcification and atherosclerotic plaque stability in carotid arteries. METHODS AND RESULTS: To this end, we characterized the calcification on 229 carotid plaques, by morphology, immunohistochemistry, transmission electron microscopy and energy dispersive X-ray microanalysis. Plaques were classified into two categories: unstable and stable. No significant differences were found in the incidence of the various risk factors between patients with and without carotid calcification, with the exception of diabetes. The energy dispersive X-ray microanalysis allowed us to identify two types of calcium salts in the atheromatous plaques, hydroxyapatite (HA) and calcium oxalate (CO). Our results showed that calcification is a common finding in carotid plaques, being present in 77.3% of cases, and the amount of calcium is not a factor of vulnerability. Noteworthy, we observed an association between HA calcification and unstable plaques. On the contrary, CO calcifications were mainly detected in stable plaques. CONCLUSIONS: The presence of different types of calcification in atheromatous plaques may open new perspectives in understanding the molecular mechanisms of atheroma formation and plaque instability.

Elemental analysis of histological specimens: a method to unmask nano asbestos fibers.

There is recent mounting evidence that nanoparticles may have enhanced toxicological potential in comparison to the same material in the bulk form. The aim of this study was to develop a new method for unmask asbestos nanofibers from Formalin-Fixed, Paraffin-Embedded tissue. There is an increasing amount of evidence that nanoparticles may enhance toxicological potential in comparison to the same material in the bulk form. The aim of this study was to develop a new method to unmask asbestos nanofibers from Formalin-Fixed Paraffin-Embedded (FFPE) tissue. For the first time, in this study we applied Energy Dispersive X-ray (EDX) microanalysis through transmission electron microscopy to demonstrate the presence of asbestos nanofibers in histological specimens of patients with possible occupational exposure to asbestos. The diagnostic protocol was applied to 10 randomly selected lung cancer patients with no history of previous asbestos exposure. We detected asbestos nanofibers in close contact with lung cancer cells in two lung cancer patients with previous possible occupational exposure to asbestos. We were also able to identify the specific asbestos iso-type, which in one of the cases was the same rare variety used in the workplace of the affected patient. By contrast, asbestos nanofibers were not detected in lung cancer patients with no history of occupational asbestos exposure. The proposed technique can represent a potential useful tool for linking the disease to previous workplace exposure in uncertain cases. Furthermore, Formalin-Fixed Paraffin-Embedded (FFPE) tissues stored in the pathology departments might be re-evaluated for possible etiological attribution to asbestos in the case of plausible exposure. Since diseases acquired through occupational exposure to asbestos are generally covered by workers' insurance in most countries, the application of the protocol used in this study may have also relevant social and economic implications.

A fatal case of invasive fungal sinusitis by Scopulariopsis acremonium in a bone marrow transplant recipient.

A fatal case of Scopulariopsis acremonium sinus infection in an allogeneic hematopoietic stem cell transplant patient is reported. Rapid vascular diffusion of the fungus to the major head vessels was observed, which led to subsequent repeated cerebral ischemia and death. The presence of hyphae in the right carotid wall might be considered an indirect sign of fungal blood diffusion in the absence of positive blood cultures. The infection developed during the course of prolonged voriconazole prophylaxis, which was found to be effective in the in vitro antifungal drug assay. This finding induced us to consider the capacity of this drug to reach infected paranasal sinuses, and the need in cases such as this of a combined systemic and local pharmacological therapy or a combined medical and surgical approach.

Small bowel adenocarcinoma as first presentation of coeliac disease.

Coeliac disease is a chronic inflammatory disease of the gut with increased risk of gastrointestinal malignancy. Although enteropathy T-cell lymphoma is the most common neoplasm in coeliacs, an increased frequency of small bowel carcinoma has been described. A case is described of jejunal carcinoma as first presentation of coeliac disease, in which gastrointestinal and extraintestinal symptoms of disease developed only after surgical resection and disappeared after gluten withdrawal.

Flow cytometry analysis of atherosclerotic plaque cells from human carotids: a validation study.

BACKGROUND: Atherosclerotic plaques are heterogeneous vascular lesions. Changes in cell plaque composition are fundamental events inside the plaque microenvironment that are strictly related to the clinical outcome of these lesions (organ damage). The knowledge of these modifications may help to better understand the pathophysiological mechanisms of atherosclerosis. METHODS: We report on a flow cytometry method to characterize and quantify the cell subpopulations in human atherosclerotic plaques. Cells were obtained from endarterectomy specimens after collagenase digestion. Both surface and intracytoplasmic antigens were labeled. RESULTS: Our data demonstrated that the method we described allowed the characterization of cell populations that compose the atherosclerotic plaque, avoiding contamination by tunica media smooth muscle cells and the noise of cellular debris. Moreover this validation study showed that about 50% of cells in the atherosclerotic plaques are inflammatory mononuclear cells (T lymphocytes and monocytes/macrophages). CONCLUSIONS: Reproducible quantitative methods for cell population characterization may increase the understanding of pathophysiological mechanisms responsible for plaque progression. The methodology herein described gave us the possibility of quickly calculating the relative amount of each cell population and studying both surface and intracellular markers to analyze the functional stage of the cells. The clinical correlation was not assessed in the present study, because we used a small patient group to validate the method, but should be the subject of further analyses in a larger patient population.

[Metaplastic tumors of the breast: a case of primary squamous cell carcinoma]. / Tumori metaplasici della mammella: un caso di carcinoma squamoso primitivo.

We report a case of primary squamous cell carcinoma of the breast in a 58 years old woman. The diagnosis of this rare tumor is possible after excluding a skin primary lesion or an epidermoid cancer of a distant site. In reviewing the reported cases we didn't find any significant prognostic difference between this form and the breast adenocarcinoma with squamous metaplasia.

Densitometric and morphometric study of immunocytochemical estrogen receptors detection in breast carcinomas.

Immunohistochemical quantitative evaluation of estrogen receptors (ER) detected in tissue sections from 30 breast tumors by monoclonal antibody was performed using a densitometric method. In particular, ER concentration was calculated by nuclear mean optical density (nMOD), while heterogeneity in ER content was calculated by the coefficient of variation (CV) of the nuclear optical density histogram. Tumors which showed more than 60% of positive cells had a mean value of ER-nMOD of 0.116 +/- 0.002 a.u. and of ER-CV of 33.74 +/- 0.68. Tumors which showed 30% to 60% of positive cells had a mean value of ER-nMOD of 0.082 +/- 0.006 a.u. (arbitrary units) and of ER-CV of 36.25 +/- 3.44. Tumors showing less than 30% of positive cells had ER-nMOD of 0.052 +/- 0.009 a.u. and ER-CV of 48.49 +/- 5.61. These results indicate that the greater the concentration the lower the ER heterogeneity within the tumor sample. No significant differences between ER-ICA results, nuclear size and form factors were found.

Carcinoma of the vulva with sarcomatoid features: a case report with immunohistochemical study.

A tumor of the vulva with sarcomatoid features was studied by immunocytochemistry to characterize the phenotype of the spindle-shaped and giant cells. Sarcomatoid-looking cells were positive for intermediate filament keratin polypeptides of stratified epithelium. These results favor histogenesis of the sarcomatoid-looking cells from a metaplastic alteration of the malignant squamous component.

Carbohydrate histochemistry of human Brunner's glands.

Brunner's glands are known to produce neutral mucins. In order to achieve a better knowledge of their carbohydrate profile, we used five peroxidase-labeled lectins on surgical specimens of human duodenum. This method allowed us to identify at least two different types of neutral mucins in Brunner's glands secretion, thus demonstrating a heterogeneous mucin production. The structure of terminal oligosaccharidic chains in these glycoproteins has also been hypothesized.