Anna Martorana

BRAFV600E mutation and p27kip1 expression in papillary carcinomas of the thyroid ≤1 cm and their paired lymph node metastases

BRAFV600E mutation and p27kip1 expression have been introduced as novel indicators that may predict prognosis in different tumors, as well as in papillary thyroid carcinomas.

Nanostructured Lipid Carriers-Containing Anticancer Compounds: Preparation, Characterization, and Cytotoxicity Studies

This article describes the development of nanostructured lipid carriers (NLC) as colloidal carriers for two antitumor compounds that possess a remarkable antineoplastic activity. But their limited stability and low solubility in water could give a very low parenteral bioavailability. Results revealed an enhancement of the cytotoxicity effect of drug-loaded NLC on human prostate cancer (PC-3) and human hepatocellular carcinoma (HuH-6, HuH-7) cell lines with respect to that of both free drugs. Results of characterization studies strongly support the potential application of these drugs-loaded NLC as prolonged delivery systems for lipophilic drugs by several administration routes, in particular for intravenous administration.

BRAF(V600E) mutation influences hypoxia-inducible factor-1alpha expression levels in papillary thyroid cancer.

Hypoxia-inducible factor-1α is found frequently overexpressed in solid tumors cells, exerting an important role in angiogenesis, glucose metabolism, cell proliferation, survival and invasion. In thyroid carcinomas, hypoxia-inducible factor-1α expression was found increased in differentiated, poorly differentiated, medullary and anaplastic variants. Hypoxia represents the principal stimulus responsible for hypoxia-inducible factor-1α induction. Other nonhypoxic stimuli increase hypoxia-inducible factor-1α synthesis through the activation of phosphatidylinositol 3-kinase and mitogen-activated protein kinase pathways in a cell-type-specific manner. We have previously shown the role of BRAFV600E mutation in papillary thyroid cancer cells as a factor that facilitates tumor cell growth and progression. In this study, we tested the hypothesis that BRAFV600E mutation influences hypoxia-inducible factor-1α expression in papillary thyroid carcinoma cells. We analyzed 27 papillary thyroid carcinomas, 13 of which presented BRAFV600E mutation. In tumor tissues, immunoreactivity for hypoxia-inducible factor-1α was detected in the majority of analyzed BRAFV600E mutated cases. Transcriptional analyses revealed elevated hypoxia-inducible factor-1α levels with significant differences between wild-type and mutated group. A BRAF wild-type papillary thyroid carcinoma cell line and a BRAFV600E mutated papillary thyroid carcinoma cell line were selected to study the effects of BRAF mutation on hypoxia-inducible factor-1α expression in vitro. Knockdown of mutant BRAFV600E or both the wild type and the BRAFV600E by RNA interference induced a significant reduction of hypoxia-inducible factor-1α expression at mRNA and protein levels. Pharmacological inhibition of BRAF significantly reduces hypoxia-inducible factor-1α expression levels in papillary thyroid carcinoma cell line harboring BRAFV600E mutation. Our results suggest that hypoxia-inducible factor-1α is expressed in papillary thyroid carcinomas and is regulated not only by hypoxia but also by BRAFV600E-mediated signaling pathway.

Hsp60 and Hsp10 increase in colon mucosa of Crohn’s disease and ulcerative colitis

The purpose of this work was to determine in colon mucosa of Crohn’s disease (CD) and ulcerative colitis (UC) in relapse: a) the levels of the chaperonins Hsp60 and Hsp10; b) the quantity of inflammatory cells; and c) if the levels of chaperonins parallel those of inflammation cells. Twenty cases of CD and UC and twenty normal controls (NC) were studied using immunohistochemistry, Western blotting and immunofluorescence. Immunohistochemically, Hsp60 and Hsp10 were increased in both inflammatory bowel diseases (IBD) compared to NC. These results were confirmed by Western blotting. Hsp60 and Hsp10 occurred in the cytoplasm of epithelial cells in CD and UC but not in NC. Hsp60 and Hsp10 co-localised to epithelial cells of mucosal glands but not always in connective tissue cells of lamina propria, where only Hsp60 or, less often, Hsp10 was found. Cells typical of inflammation were significantly more abundant in CD and UC than in NC. Since chaperonins are key factors in the activation of the immune system leading to inflammation, we propose that they play a central role in the pathogenesis of the two diseases, which, consequently, ought to be studied as chaperonopathies.

Increased expression of transketolase‐like‐1 in papillary thyroid carcinomas smaller than 1.5 cm in diameter is associated with lymph‐node metastases

Patients with small papillary thyroid carcinoma (PTC) may have a high incidence of regional lymph‐node (LN) metastases at presentation, and these are considered to be an independent risk factor for tumor recurrence. A mutated transketolase transcript (TKTL1) has been found up‐regulated in different human malignancies, and strong TKTL1 protein expression has been associated with aggressiveness and poor patient survival in several epithelial cancers.

Associations between Notch-2, Akt-1 and HER2/neu expression in invasive human breast cancer: a tissue microarray immunophenotypic analysis on 98 patients.

Objective: We aimed to investigate the existence of associations between well-established and newly recognized biological and phenotypic features of breast cancer involved in tumor progression and prognosis. Methods: Ninety-eight cases of invasive breast cancer were assessed for the immunohistochemical expression of estrogen and progesterone receptors, Ki-67, HER2, Akt-1, and Notch-2, using the tissue microarray technique. Data regarding tumor histotype, histological grade, tumor size and lymph node status were collected for each patient and included in the analysis. Results: Several significant associations between histological and/or immunophenotypic features came from the analysis of our data. Positive associations were observed between estrogen and progesterone receptors, tumor grade and proliferation index, tumor grade and HER2, Akt-1 and estrogen receptors, and Notch-2 and HER2. Inverse associations were noted between hormone receptors and tumor grade, hormone receptors and HER2, Akt-1 and tumor grade, and Akt-1 and nodal invasion. Conclusions: Our results, showing the existence of a number of estrogen receptor-positive tumors with Akt-1 expression, better degree of differentiation, and no lymph node involvement, along with the presence of HER2-positive tumors with strong Notch-2 expression, support the role of Notch and Akt in breast cancer progression and suggest that they may also represent new appealing therapeutic targets.

Rare localization of low-grade fibromyxoid sarcoma to the nail region.

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Predictive role of histological features and Ki67 pattern on high-risk HPV presence in atypical cervical lesions.

the Laboratory of HealthCare Associated Infection, Specialist and Reference Microbiology Division, Colindale, UK, demonstrated S. aureus positive for the PVL gene. Molecular studies by polymerase chain reaction on the postmortem tissue were negative for all influenza, Para influenza, respiratory syncytial and adeno-viruses. Antemortem serological tests for Legionella, mycoplasma, Q fever and psittacosis were also negative. It was concluded that this was haemorrhagic necrotizing pneumonia occurring in a young immunocompetent patient due to S. aureus strain carrying the PVL gene. This was regarded as ‘community acquired’ in our case, as the patient already had established features of pneumonia and septicaemia on admission. Epidemiological evidence so far also suggests that this PVL strain of MRSA is community acquired in most circumstances. As the patient was not immunocompromised, her neutropenia appeared to be the effect of the leukocytotoxic PVL toxin as well as secondary to septicaemia and DIC. Staphylococcus aureus is responsible for about 2% of cases of community-acquired pneumonia and at least 10% of cases of nosocomial pneumonia. Staphylococcus aureus carrying the PVL gene strain is responsible for the community-acquired pneumonias. CA-MRSA has several distinguishing features from hospital acquired (HA)-MRSA, including: presence of the SCCmec IV element and the presence of a gene encoding the PVL toxin; they are more virulent, although relatively sensitive to antibiotics. Depending on multi-locus sequence typing and pulsed-field gel electrophoresis, Vandenesch et al. and Charlebois et al. have concluded that the genetic substance of CA-MRSA and HA-MRSA is diverse and distinct, indicating that CA-MRSA are true strains originating from the community. Therefore, PVL-MRSA ⁄ CAMRSA seems to be a specific disease entity with a poor prognosis. This poor prognosis is probably related to the PVL toxin, which is an extracellular product with a leucocytotoxic action. The prevalence of CA-MRSA is not known, but it can be distinguished from the nosocomial strains clinically by its susceptibility profile (Ciprofloxacin susceptible, fusidic acid resistant). The pathological findings in this case are similar to the (limited) pathology findings described in the literature. The gross appearance of the lungs is similar to either viral pneumonia or adult respiratory distress syndrome, and is different from suppurative pneumonia due to other bacterial infections. Histological features are also distinctly different from other forms of staphylococcal pneumonia with no evidence of suppuration. They more resemble viral pneumonia, except, of course, for the lack of viral inclusions and the presence of large colonies of Gram-positive cocci. Although PVL gene-carrying MRSA pneumonia ⁄ CA-MRSA pneumonia has been reported and recognized in the literature, it is not widely recognized by histopathologists and the detailed pathology of lungs has not been well described. This has important implications. CA-MRSA is likely to present as an acute fulminating illness in previously healthy immunocompetent young adults, culminating in death (within a short period of time). As histopathologists are likely to be performing autopsies in such cases, lack of knowledge of the existence (and pathology) of CA-MRSA ⁄ PVL-MRSA could result in an erroneous diagnosis and cause of death.

Difference in Ki67 and Thymidylate Synthase Expression in Primary Tumour Compared with Metastatic Nodes in Breast Cancer Patients

Breast cancer is a heterogeneous disease, so therapeutic predictive biological markers need to be identified. To date an accurate evaluation of predictive markers is mainly done at the primary site; however, the main goal of adjuvant therapy for breast cancer is the control of micrometastases. The aim of this study is to assess as therapeutic and/or prognostic marker, the proliferation status of primary tumors and involved nodes as measured by Ki67 and thymidylate synthase (TS) expression, in 30 breast cancer node positive patients. TS is the main target of 5-fluorouracil (5-FU) activity, and its overexpression is one of the mechanisms of 5-FU drug resistance; however, in some studies its absence is responsible for a worse response to 5-FU. Our results show that malignant cells of involved nodes were in a post mitotic phase of the cell cycle, and show a low proliferation index and TS expression, while the primary tumours and controls, were strongly positive. On these basis we can hypothesize that these cells could be less sensitive to 5-FU. Further studies are necessary to identify other mechanisms responsible for their metastasing capability and/or for their aggressiveness.

Seeding nerve sutures with minced nerve-graft (MINE-G): a simple method to improve nerve regeneration in rats

Abstract Background: The aim of this study was to assess the effect of seeding the distal nerve suture with nerve fragments in rats. Methods: On 20 rats, a 15 mm sciatic nerve defect was reconstructed with a nerve autograft. In the Study Group (10 rats), a minced 1 mm nerve segment was seeded around the nerve suture. In the Control Group (10 rats), a nerve graft alone was used. At 4 and 12 weeks, a walking track analysis with open field test (WTA), hystomorphometry (number of myelinated fibers (n), fiber density (FD) and fiber area (FA) and soleus and gastrocnemius muscle weight ratios (MWR) were evaluated. The Student t-test was used for statistical analysis. Results: At 4 and 12 weeks the Study Group had a significantly higher n and FD (p = .043 and .033). The SMWR was significantly higher in the Study Group at 12 weeks (p = .0207). Conclusions: Seeding the distal nerve suture with nerve fragments increases the number of myelinated fibers, the FD and the SMWR. The technique seems promising and deserves further investigation to clarify the mechanisms involved and its functional effects.