CMA mediates resistance in breast cancer models.

BACKGROUND: Breast cancer (BC) is the most common malignancy in women and the second leading cause of cancer-related death; chemoresistance is still a clinical challenge mainly because of the different molecular features of this kind of tumour. Doxorubicin (Doxo) is widely used despite its adverse effects and the common onset of resistance. Chaperone-Mediated Autophagy (CMA) has been identified as an important mechanism through which chemotherapeutics can exert their cytotoxic effects and, in this context, LAMP-2A, the key player of CMA, can be a useful biomarker. METHODS: A cohort of patients and breast cancer cells have been screened for Doxo effect and CMA activation by analysing the LAMP-2A level. Molecular silencing has been used to clarify CMA role in BC responsiveness to treatments. Low Doxo doses were combined with other drugs (TMZ or PX-478, a HIF-1α inhibitor) to evaluate their cytotoxic ability and their role in modulating CMA. RESULTS: In this paper, we showed that CMA is an important mechanism mediating the responsiveness of breast cancer cell to different treatments (Doxo and TMZ, as suggested by triple negative cells that are TMZ-resistant and fails to activate CMA). The LAMP-2A expression level was specific for different cell lines and patient-derived tumour subtypes, and was also useful in discriminating patients for their survival rates. Moreover, molecular silencing or pharmacological blockage of HIF-1α activity reverted BC resistance to TMZ. The combination of low-dose Doxo with TMZ or PX-478 showed that the drug associations have synergistic behaviours. CONCLUSION: Here, we demonstrated that CMA activity exerts a fundamental role in the responsiveness to different treatments, and LAMP-2A can be proposed as a reliable prognostic biomarker in breast cancer. In this context, HIF-1α, a potential target of CMA, can also be assessed as a valuable therapeutic target in BC in view of identifying new, more efficient and less toxic therapeutic drug combinations. Moreover, the possibility to combine Doxo with other drugs acting on different but coherent molecular targets could help overcome resistance and open the way to a decrease in the dose of the single drugs.

Finite-Temperature Disordered Bosons in Two Dimensions.

We study phase transitions in a two dimensional weakly interacting Bose gas in a random potential at finite temperatures. We identify superfluid, normal fluid, and insulator phases and construct the phase diagram. At T=0 one has a tricritical point where the three phases coexist. The truncation of the energy distribution at the trap barrier, which is a generic phenomenon in cold atom systems, limits the growth of the localization length and in contrast to the thermodynamic limit the insulator phase is present at any temperature.

Diagnostic Circulating miRNAs in Sporadic Amyotrophic Lateral Sclerosis.

Amyotrophic Lateral Sclerosis (ALS) is a fatal neurodegenerative disease characterized by the neurodegeneration of motoneurons. About 10% of ALS is hereditary and involves mutation in 25 different genes, while 90% of the cases are sporadic forms of ALS (sALS). The diagnosis of ALS includes the detection of early symptoms and, as disease progresses, muscle twitching and then atrophy spreads from hands to other parts of the body. The disease causes high disability and has a high mortality rate; moreover, the therapeutic approaches for the pathology are not effective. miRNAs are small non-coding RNAs, whose activity has a major impact on the expression levels of coding mRNA. The literature identifies several miRNAs with diagnostic abilities on sALS, but a unique diagnostic profile is not defined. As miRNAs could be secreted, the identification of specific blood miRNAs with diagnostic ability for sALS could be helpful in the identification of the patients. In the view of personalized medicine, we performed a meta-analysis of the literature in order to select specific circulating miRNAs with diagnostic properties and, by bioinformatics approaches, we identified a panel of 10 miRNAs (miR-193b, miR-3911, miR-139-5p, miR-193b-1, miR-338-5p, miR-3911-1, miR-455-3p, miR-4687-5p, miR-4745-5p, and miR-4763-3p) able to classify sALS patients by blood analysis. Among them, the analysis of expression levels of the couple of blood miR-193b/miR-4745-5p could be translated in clinical practice for the diagnosis of sALS.

Distinct populations of tumor-initiating cells derived from a tumor generated by rat mammary cancer stem cells.

Tumors derived from rat LA7 cancer stem cells (CSCs) contain a hierarchy of cells with different capacities to generate self-renewing spheres and tubules serially ex vivo and to evoke tumors in vivo. We isolated two morphologically distinct cell types with distinct tumorigenic potential from LA7-evoked tumors: cells with polygonal morphology that are characterized by expression of p21/(WAF1) and p63 and display hallmarks of CSCs and elongated epithelial cells, which generate tumors with far less heterogeneity than LA7 CSCs. Serial transplantation of elongated epithelial cells results in progressive loss of tumorigenic potential; tumor heterogeneity; CD44, E-cadherin, and epithelial cytokeratin expression and increased alpha-smooth muscle actin I and vimentin expression. In contrast, serial transplantation of LA7 CSCs can be performed indefinitely and results in tumors that maintain their heterogeneity, consistent with self-renewal and multilineage differentiation potential. Collectively, our data show that polygonal cells are CSCs, whereas epithelial elongated cells are lineage-committed progenitors with tumorigenic potential, and suggest that tumor progenitors, although lacking indefinite self-renewal potential, nevertheless may make a substantial contribution to tumor development. Because LA7 cells can switch between conditions that favor maintenance of pure CSCs vs. differentiation into other tumor cell types, this cell system provides the opportunity to study factors that influence CSC self-renewal and differentiation. One factor, p63, was identified as a key gene regulating the transition between CSCs and early progenitor cells.

Isolation of canine mammary cells with stem cell properties and tumour-initiating potential.

Recent data suggest that mammary carcinogenesis may be driven by cancer stem cells (CSCs) derived from mutated adult stem cells, which have acquired aberrant cell self-renewal or by progenitor cells that have acquired the capacity for cell self-renewal. Spontaneous mammary cancers in cats and dogs are important models for the understanding of human breast cancer and may represent alternative species model systems that can significantly contribute to the study of human oncogenesis. With the goal of identifying markers for isolating human breast CSCs, we have generated a canine model system to isolate and characterize normal and CSCs from dog mammary gland. Insight into the hierarchical organization of canine tumours may contribute to the development of universal concepts in oncogenesis by CSCs. Cells with stem cell properties were isolated from normal and tumoural canine breast tissue and propagated as mammospheres and tumourspheres in long-term non-adherent culture conditions. We showed that cells obtained from spheres that display self-renewing properties, have multi-lineage differentiation potential, could generate complex branched tubular structures in vitro and form tumours in NOD/SCID mice. We analysed these cells for the expression of human stem and CSC markers and are currently investigating the tumour-initiating properties of these cells and the hierarchical organization of normal and neoplastic canine mammary tissue.

[An unusual localization of thryoglossal duct cyst]. / Localizzazione inusuale di cisti del dotto tireoglosso.

Thyroglossal duct cysts are most common neck masses after benign lymphonodes. They originate from primitive thiroglossal duct, so they could be locate along its course. Every mass in the middle line of the neck can be considered as a thyroglossal cyst. Best treatment is surgery (Sistrunk procedure). We present a case of unusual localization at floor of the mouth of thyroglossal in a 34 years old woman. To our knowledge in literature, only two cases, have been reported both.

p53 but not bcl-2 immunostaining is predictive of poor clinical complete response to primary chemotherapy in breast cancer patients.

Preoperative chemotherapy administered to breast cancer (BC) patients is a model for studying in vivo the interaction between cytotoxic treatment and clinical and biological parameters. Apoptosis induced by anticancer agents is a mechanism of treatment activity; therefore, overexpression of genes inhibiting the apoptotic pathway could produce drug resistant tumors. In the present study, the two most studied inhibitors of apoptosis, the bcl-2 gene and the mutant p53, have been evaluated to assess whether they may play a role in modulating response of BC to primary chemotherapy. From August 1990 to January 1997, 143 patients bearing T(2-4)N(0-1)M0 primary BC were submitted to two different chemotherapeutic regimens before surgery. The first 64 received the cyclophosphamide, methotrexate, 5-fluorouracil (CMF) regimen (on days 1 and 8 and every 28 days thereafter) associated with tamoxifen (30 mg daily) in case of estrogen receptor (ER)-positive BC, and the remaining 79 were submitted to single agent epirubicin (120 mg/m2 every 21 days). The expression of p53, bcl-2, Ki67, ER, progesterone receptor, c-erbB2, and the multidrug resistance P-glycoprotein (gp-170) was evaluated in BC specimens obtained at diagnosis by incision biopsy and at postchemotherapy surgery. At the end of chemotherapy administration (median, 3 cycles; range, 2-6), the clinical complete response (cCR) rate was superimposable in the patient subgroups with bcl-2-positive or -negative primary tumors; conversely, p53 expression, at a cutoff of 10% positive cells, was significantly associated with a lower cCR rate (9.4 versus 27.0%; P < 0.04). p53 was a significant predictor for poor cCR in the subset submitted to epirubicin (3.6 versus 25.5%; P < 0.02; in patients with p53+ and p53- BC, respectively); by contrast, only a trend toward lower cCR has been observed in patients with p53+ tumors receiving CMF +/- tamoxifen with respect to p53- ones. The distribution of cCR according to the gp-170-positive or -negative tumors was 8 versus 22% in patients submitted to epirubicin and 29 versus 30% in those receiving CMF +/- tamoxifen, respectively. In a multivariate regression analysis, after adjusting for treatment administered (epirubicin versus CMF +/- tamoxifen), menopausal status, tumor and node status, histology grade, ER, progesterone receptor, c-erbB2, Ki67, bcl-2, and gp-170 expression, the p53 status maintained an independent predictive role for cCR. Most of the tumors undergoing change in percentage of p53 expression after both treatments originally harbored mutant protein, and only four BC specimens that were p53 negative before chemotherapy became positive afterward. These data confirm in vivo the concept that the responsiveness of tumors to chemotherapy in part derives from the capability of BC cells to undergo apoptosis. The role of mutated p53 in preventing response is more evident in patients submitted to epirubicin, and this may be caused by the up-regulation of multidrug resistance gene expression by p53 inactivation. p53 is a stable phenotype and is not inducible by at least three or four chemotherapy cycles.

The C-terminal domain of yeast Ero1p mediates membrane localization and is essential for function.

In eukaryotes, members of the Ero1 family control oxidative protein folding in the endoplasmic reticulum (ER). Yeast Ero1p is tightly associated with the ER membrane, despite cleavage of the leader peptide, the only hydrophobic sequence that could mediate lipid insertion. In contrast, human Ero1-Lalpha and a yeast mutant (Ero1pDeltaC) lacking the 127 C-terminal amino acids are soluble when expressed in yeast. Neither Ero1-Lalpha nor Ero1pDeltaC complements an ERO1 disrupted strain. Appending the yeast C-terminal tail to human Ero1-Lalpha restores membrane association and allows growth of ERO1 disrupted cells. Therefore, the tail of Ero1p mediates membrane association and is crucial for function.

Effect of neoadjuvant chemotherapy on Ki67 labelling index, c-erbB-2 expression and steroid hormone receptor status in human breast tumours.

The administration of neoadjuvant chemotherapy to breast cancer (BC) patients with operable disease allowed studies aimed of exploring the interaction between cytotoxic treatment and tumour biology in vivo. 99 patients with T2-4, NO-1, M0 primary BC received a median of 3 cycles of either CMF regimen (cyclophosphamide, methotrexate, 5-fluorouracil) or single agent epirubicin. Endocrine therapy was also concomitantly administered in the first 45 patients with estrogen receptor positive (ER+) BC. 92 ended the treatment plan. Ki67 labelling index, estrogen receptor (ER), progesterone receptor (PgR), and c-erbB-2 oncoprotein expression were evaluated immunohistochemically in tumour biopsies obtained before and after chemotherapy. At post-chemotherapy evaluation, tumour shrinkage greater than 50% was obtained in 71 patients (79.7%), 27 of them being complete responders (30.3%). The median Ki67 labelling index, which was 13% in the first biopsy, decreased to 4.5% (p < 0.001) upon mastectomy. No significant differences were observed in steroid hormone receptor and c-erbB-2 expression before and after neoadjuvant treatment. In conclusion, neoadjuvant chemotherapy, whether associated or not to endocrine therapy, leads to a significant decrease in BC proliferation without any appreciable impact on c-erbB-2 and steroid hormone receptor expression.

Video-based system for intraoperative facial nerve monitoring: comparison with electromyography.

OBJECTIVE: To validate a recently developed intraoperative facial nerve monitoring system that is based on video control of facial movements. STUDY DESIGN: In a single-subject design study, involving 15 otoneurosurgical patients, the relationship between intensity of neural stimulation, facial movements, and electrophysiologic voltage were measured. The analysis was performed by measuring the ipsilateral oral commissure displacement in relation to different levels of current administered to the nerve during surgical procedures. SETTING: Electromyography and video system intraoperative facial nerve monitoring. PATIENTS: 15 patients (9 men, 6 women; mean age, 61 yr) undergoing a translabyrinthine approach for removal of acoustic neuroma. RESULTS: Electromyography showed slightly greater sensitivity. With regard to the stimulation-response ratio, facial movement and electromyographic amplitude showed very similar responses. CONCLUSIONS: The video system was considered useful in terms of validity and reliability. Furthermore, the authors' surgical experience showed some limitations of electromyography, especially in terms of electrical artifact during cauterization, totally masking the electrophysiologic monitoring.

[Role of platelet serotonin in hyperperistalsis after digestive hemorrhages]. / Ruolo della serotonina piastrinica nell'iperperistaltismo dopo emorragie digestive.

Intestinal transit accelerates during gastroduodenal haemorrhage. To test a personal theory that the reason is to be found in serotonin released by platelets and absorbed by the intestine, the behaviour of serotoninaemia has been studied. This proved to be 1.35-9.7 microgram/l (mean: 4.175 microgram/l; E.S. +/- 1.39) in patients with haematemesis and melena, compared to 0.49-0.9 microgram/l (mean: 0.728 microgram/l; E.S. +/- 0.07) in the controls (p less than 0.001). The increase in circulating serotonin to significant values is probably due to hormone release. This dose not occur in cases of melena alone, probably owing to the organism's high capacity for catabolism.

[Primary lung neoplasm of rare incidence: giant cell carcinoma]. / Una neoplasia primitiva del polmone di non frequente riscontro: il carcinoma a cellule giganti

Primary giant-cell carcinoma of the lung is an infrequent observation. It is distinguished clinically solely its fairly rapid development but otherwise it does not seem to have any predilection for localization, age or sex. Histologically, differential diagnosis with othe tumours such as angiosarcoma, rhabdomyosarcoma, chorioepithelioma and others is of interest. Three cases of primary giant-cell carcinoma of the lung are reported and the various clinical and anatomopathological aspects discussed.

Sudden improvement of respiratory failure in a woman with carcinoma of the colon.

We present a case of bronchial and endotracheal metastases completely blocking the left main bronchus and partially occluding the middle lobe bronchus contributing to severe respiratory failure. The patient's lack of consent to laser resection of the mass led to the use of chemotherapy; after the first cycle of treatment a neoplastic mass about 3 cm long was spontaneously expelled with a cough. The expulsion of the metastasis caused rapid improvement of the dyspnea and gas exchange; however, the continuation of the chemotherapy did not bring any further benefit to the patient, who died 115 days after diagnosis.

Initial experience of peroperative electrocochleography monitoring in stapedectomy.

Clinical experience of intraoperative ECochG monitoring during stapes surgery is reported. Selected surgical phases have been taken into consideration, laying particular stress on the phase corresponding to the opening of the labyrinthine spaces. Compound action potential (CAP) amplitude and latency showed, a supposed, an inversely proportional relationship in most of the cases. Abnormal findings were also recorded in some patients, thus giving rise to different electrophysiological hypotheses. A proposal for an "in vivo" model for the study of the electrophysiological changes due to the opening of labyrinthine spaces is made.

[An unusual case of hematuria]. / Un caso insolito di ematuria.

The author describes a unusual case of haematuria from Schistosoma Haematobium and dispute the modalities of diagnosis and therapy.

[A case of transient congenital hypoaldosteronism]. / Un caso di ipoaldosteronismo congenito transitorio.

A case of temporary congenital hypoaldosteronism with normal production of other hormones of the adrenal is described. During early infancy the patient showed low blood aldosterone levels, increased ACHT and Hyponatraemia. Replacement therapy needed sodium chloride and mineralcorticoid drug administration (Florinef) until the baby was 9 month old. Later, spontaneous recovery occurred, confirmed by laboratory tests.

[Visceral leishmaniasis. Practical considerations on a clinical case]. / La Leishmaniosi viscerale. Considerazioni pratiche su un caso clinico.

A male infant with profound anemia and liver and spleen enlargement was affected by visceral Leishmaniasis. Diagnostic tests and therapeutical approach to this parasitic infection, very rare in our Region, are discussed.

Pathway-based expression profile for breast cancer diagnoses.

Microarray experiments have made possible to identify breast cancer marker gene signatures. However, gene expression-based signatures present limitations because they do not consider metabolic role of the genes and are affected by genetic heterogeneity across patient cohorts. Considering the activity of entire pathways rather than the expression levels of individual genes can be a way to exceed these limits. We evaluated and compared five methods of pathway-level aggregation of gene expression data. Our results confirmed the important role of pathway expression profile in breast cancer diagnostic classification (accuracy >90%). However, although assessed on a limited number of samples and datasets, this study shows that using dissimilarity representation among patients does not improve the classification of pathway-based expression profiles.