The Novel Role of SOX2 as an Early Predictor of Cancer Risk in Patients with Laryngeal Precancerous Lesions.

The SOX2 gene located at 3q26 is frequently amplified and overexpressed in multiple cancers, including head and neck squamous cell carcinomas (HNSCC). The tumor-promoting activity and involvement of SOX2 in tumor progression has been extensively demonstrated, thereby emerging as a promising therapeutic target. However, the role of SOX2 in early stages of tumorigenesis and its possible contribution to malignant transformation remain unexplored. This study investigates for the first time SOX2 protein expression by immunohistochemistry and gene amplification by real-time PCR using a large series of 94 laryngeal precancerous lesions. Correlations with the histopathological classification and the risk of progression to invasive carcinoma were established. Nuclear SOX2 expression was frequently detected in 38 (40%) laryngeal dysplasias, whereas stromal cells and normal adjacent epithelia showed negative expression. SOX2 gene amplification was detected in 18 (33%) of 55 laryngeal dysplasias. Univariate Cox analysis showed that SOX2 gene amplification (p = 0.046) and protein expression (p < 0.001) but not histological grading (p = 0.432) were significantly associated with laryngeal cancer risk. In multivariate stepwise analysis including age, tobacco, histology, SOX2 gene amplification and SOX2 expression, SOX2 expression (HR = 3.531, 95% CI 1.144 to 10.904; p = 0.028) was the only significant independent predictor of laryngeal cancer development. These findings underscore the relevant role of SOX2 in early tumorigenesis and a novel clinical application of SOX2 expression as independent predictor of laryngeal cancer risk in patients with precancerous lesions beyond current WHO histological grading. Therefore, targeting SOX2 could lead to effective strategies for both cancer prevention and treatment.

Frequent Alteration of Annexin A9 and A10 in HPV-Negative Head and Neck Squamous Cell Carcinomas: Correlation with the Histopathological Differentiation Grade.

The annexin protein superfamily has been implicated in multiple physiological and pathological processes, including carcinogenesis. Altered expression of various annexins has frequently been observed and linked to the development and progression of various human malignancies. However, information is lacking on the expression and clinical significance of annexin A9 (ANXA9) and A10 (ANXA10) in head and neck squamous cell carcinomas (HNSCC). ANXA9 and ANXA10 expression was evaluated in a large cohort of 372 surgically treated HPV-negative HNSCC patients and correlated with the clinicopathologic parameters and disease outcomes. Down-regulation of ANXA9 expression was found in 42% of HNSCC tissue samples, compared to normal epithelia. ANXA9 expression in tumors was significantly associated with oropharyngeal location and histological differentiation grade (P < 0.001). In marked contrast, ANXA10 expression was absent in normal epithelium, but variably detected in the cytoplasm of cancer cells. Positive ANXA10 expression was found in 64% of tumors, and was significantly associated with differentiation grade (P < 0.001), being also more frequent in oropharyngeal tumors (P = 0.019). These results reveal that the expression of both ANXA9 and ANXA10 is frequently altered in HNSCC and associated to the tumor differentiation grade, suggesting that they could be implicated in the pathogenesis of these cancers.

Diagnosis of invasive fungal disease in hospitalized patients with chronic obstructive pulmonary disease / Diagnóstico de micosis invasora en pacientes hospitalizados con enfermedad pulmonar obstructiva crónica

Background: The role of culture-independent techniques (galactomannan, (1-3)-ß-d-glucan) in the early diagnosis of invasive fungal diseases (IFD) is well assessed in hematological patients, but there are no clear conclusions in patients with chronic obstructive pulmonary disease (COPD). Aims: To study the usefulness of nonculture-based techniques in the diagnosis of IFD in COPD-patients at risk for IFD. Methods: A prospective observational study based on monitoring COPD patients at risk for IFD during 2007-2010 was carried out. The presence of galactomannan, (1-3)-ß-d-glucan and an indirect immunofluorescence of Candida albicans germ tube specific antibodies (CAGTA) were performed. Results: Among 43 COPD patients, 16 (37.2%) were diagnosed with IFD: seven cases were proven IFD (five invasive candidemia - IC, one invasive aspergillosis - IA and a rhinocerebral zygomycosis) and nine probable IFD (seven IA and two IC). In the diagnosis of IC and IA, the negative predictive value (NPV) of (1-3)-ß-d-glucan was 100%. Regarding CAGTA in IC, NPV was 96.2%. Finally, NPV of galactomannan in IA was 91.2%. The area under the ROC curve for (1-3)-ß-d-glucan in IC and for the rest of the IFD cases was 0.86 (95% CI, 0.79-0.93) and 0.60 (95% CI, 0.43-0.77), for CAGTA in IC was 0.83 (95% CI, 0.74-0.91) and for galactomannan in IA was 0.71 (95% CI, 0.56-0.85). Positive (1-3)-ß-d-glucan preceded the growth of Candida (average of 1.7 days) in blood culture. Conclusions: In COPD patients at risk for IFD the assayed techniques are especially useful to rule out the presence of IFD

Antecedentes: El papel de las técnicas independientes de cultivo [galactomanano, (1-3)-ß-D-glucano] en el diagnóstico precoz de micosis invasoras (MI) está bien establecido en pacientes hematológicos, pero no existen conclusiones claras en pacientes con enfermedad pulmonar obstructiva crónica (EPOC). Objetivos: Estudiar la utilidad de las técnicas independientes de cultivo en el diagnóstico de MI en pacientes con EPOC que corren el riesgo de contraer una MI. Métodos: Se llevó a cabo un estudio prospectivo y observacional en que se supervisaron pacientes con EPOC que corrían el riesgo de contraer una MI de 2007 a 2010. Para ello se estableció la existencia de galactomanano, (1-3)-ß-D-glucano y se realizó el ensayo CAGTA (inmunofluorescencia indirecta para determinar la existencia de anticuerpos IgG frente a antígenos de la superficie de la fase micelial de Candida albicans). Resultados: Se diagnosticaron 16MI en 43pacientes con EPOC (37,2%): siete fueron MI probadas (cinco candidemias invasoras [CI], una aspergilosis invasora [AI] y una cigomicosis rinocerebral) y nueve fueron MI probables (siete AI y dos CI). En el diagnóstico de CI y AI, el valor predictivo negativo (VPN) del (1-3)-ß-D-glucano fue del 100%. En cuanto al CAGTA en CI, el VPN fue del 96,2%. Finalmente, el VPN del galactomanano en AI fue del 91,2%. El área bajo la curva ROC (receiver operating characteristic) del (1-3)-ß-D-glucano en CI y en el resto de los casos de MI fue 0,86 (IC95%=0,79-0,93) y 0,60 (IC95%=0,43-0,77), para CAGTA en CI fue 0,83 (IC95%=0,74-0,91) y para galactomanano en AI fue 0,71 (IC95%=0,56-0,85). La positividad del (1-3)-ß-D-glucano se anticipó como media 1,7días al crecimiento de Candida en el hemocultivo. Conclusiones: En pacientes con EPOC que corren el riesgo de contraer MI, estas técnicas son muy útiles para descartar la existencia de MI

MMP-25 Metalloprotease Regulates Innate Immune Response through NF-κB Signaling.

Matrix metalloproteases (MMPs) regulate innate immunity acting over proinflammatory cytokines, chemokines, and other immune-related proteins. MMP-25 (membrane-type 6-MMP) is a membrane-bound enzyme predominantly expressed in leukocytes whose biological function has remained largely unknown. We have generated Mmp25-deficient mice to elucidate the in vivo function of this protease. These mutant mice are viable and fertile and do not show any spontaneous phenotype. However, Mmp25-null mice exhibit a defective innate immune response characterized by low sensitivity to bacterial LPS, hypergammaglobulinemia, and reduced secretion of proinflammatory molecules. Moreover, these immune defects can be tracked to a defective NF-κB activation observed in Mmp25-deficient leukocytes. Globally, our findings provide new mechanistic insights into innate immunity through the activity of MMP-25, suggesting that this proteinase could be a potential therapeutic target for immune-related diseases.

The diagnosis and treatment of dyskeratosis congenita: a review.

Dyskeratosis congenita (DC) is an inherited bone marrow failure (BMF) syndrome characterized by the classic triad of abnormal skin pigmentation, nail dystrophy, and oral leukoplakia. However, patients usually develop BMF and are predisposed to cancer, with increased risk for squamous cell carcinoma and hematolymphoid neoplasms. DC is a disease of defective telomere maintenance and is heterogeneous at the genetic level. It can be inherited in X-linked, autosomal dominant, or autosomal recessive patterns. Mutations in at least ten telomere- and telomerase-associated genes have been described in DC. There are no targeted therapies for DC and patients usually die of BMF due to a deficient renewing capability of hematopoietic stem cells. Allogeneic hematopoietic stem cell transplantation is the only curative treatment for BMF.

ATP-dependent Lon protease controls tumor bioenergetics by reprogramming mitochondrial activity.

We generated mice deficient in Lon protease (LONP1), a major enzyme of the mitochondrial quality control machinery. Homozygous deletion of Lonp1 causes early embryonic lethality, whereas its haploinsufficiency protects against colorectal and skin tumors. Furthermore, LONP1 knockdown inhibits cellular proliferation and tumor and metastasis formation, whereas its overexpression increases tumorigenesis. Clinical studies indicate that high levels of LONP1 are a poor prognosis marker in human colorectal cancer and melanoma. Additionally, functional analyses show that LONP1 plays a key role in metabolic reprogramming by remodeling OXPHOS complexes and protecting against senescence. Our findings demonstrate the relevance of LONP1 for cellular and organismal viability and identify this protease as a central regulator of mitochondrial activity in oncogenesis.

Prelamin A causes progeria through cell-extrinsic mechanisms and prevents cancer invasion.

Defining the relationship between ageing and cancer is a crucial but challenging task. Mice deficient in Zmpste24, a metalloproteinase mutated in human progeria and involved in nuclear prelamin A maturation, recapitulate multiple features of ageing. However, their short lifespan and serious cell-intrinsic and cell-extrinsic alterations restrict the application and interpretation of carcinogenesis protocols. Here we present Zmpste24 mosaic mice that lack these limitations. Zmpste24 mosaic mice develop normally and keep similar proportions of Zmpste24-deficient (prelamin A-accumulating) and Zmpste24-proficient (mature lamin A-containing) cells throughout life, revealing that cell-extrinsic mechanisms are preeminent for progeria development. Moreover, prelamin A accumulation does not impair tumour initiation and growth, but it decreases the incidence of infiltrating oral carcinomas. Accordingly, silencing of ZMPSTE24 reduces human cancer cell invasiveness. Our results support the potential of cell-based and systemic therapies for progeria and highlight ZMPSTE24 as a new anticancer target.

ATG4B/autophagin-1 regulates intestinal homeostasis and protects mice from experimental colitis.

The identification of inflammatory bowel disease (IBD) susceptibility genes by genome-wide association has linked this pathology to autophagy, a lysosomal degradation pathway that is crucial for cell and tissue homeostasis. Here, we describe autophagy-related 4B, cysteine peptidase/autophagin-1 (ATG4B) as an essential protein in the control of inflammatory response during experimental colitis. In this pathological condition, ATG4B protein levels increase in parallel with the induction of autophagy. Moreover, ATG4B expression is significantly reduced in affected areas of the colon from IBD patients. Consistently, atg4b (-/-) mice present Paneth cell abnormalities, as well as an increased susceptibility to DSS-induced colitis. atg4b-deficient mice exhibit significant alterations in proinflammatory cytokines and mediators of the immune response to bacterial infections, which are reminiscent of those found in patients with Crohn disease or ulcerative colitis. Additionally, antibiotic treatments and bone marrow transplantation from wild-type mice reduced colitis in atg4b (-/-) mice. Taken together, these results provided additional evidence for the importance of autophagy in intestinal pathologies and describe ATG4B as a novel protective protein in inflammatory colitis. Finally, we propose that atg4b-null mice are a suitable model for in vivo studies aimed at testing new therapeutic strategies for intestinal diseases associated with autophagy deficiency.

Matrix metalloproteinase Mmp-1a is dispensable for normal growth and fertility in mice and promotes lung cancer progression by modulating inflammatory responses.

Human MMP-1 is a matrix metalloproteinase repeatedly associated with many pathological conditions, including cancer. Thus, MMP1 overexpression is a poor prognosis marker in a variety of advanced cancers, including colorectal, breast, and lung carcinomas. Moreover, MMP-1 plays a key role in the metastatic behavior of melanoma, breast, and prostate cancer cells. However, functional and mechanistic studies on the relevance of MMP-1 in cancer have been hampered by the absence of an in vivo model. In this work, we have generated mice deficient in Mmp1a, the murine ortholog of human MMP1. Mmp1a(-/-) mice are viable and fertile and do not exhibit obvious abnormalities, which has facilitated studies of cancer susceptibility. These studies have shown a decreased susceptibility to develop lung carcinomas induced by chemical carcinogens in Mmp1a(-/-) mice. Histopathological analysis indicated that tumors generated in Mmp1a(-/-) mice are smaller than those of wild-type mice, consistently with the idea that the absence of Mmp-1a hampers tumor progression. Proteomic analysis revealed decreased levels of chitinase-3-like 3 and accumulation of the receptor for advanced glycation end-products and its ligand S100A8 in lung samples from Mmp1a(-/-) mice compared with those from wild-type. These findings suggest that Mmp-1a could play a role in tumor progression by modulating the polarization of a Th1/Th2 inflammatory response to chemical carcinogens. On the basis of these results, we propose that Mmp1a knock-out mice provide an excellent in vivo model for the functional analysis of human MMP-1 in both physiological and pathological conditions.

Use of a new ziprasidone-selective electrode in mixed solvents and its application in the analysis of pharmaceuticals and biological fluids.

The construction and characterization of a new ion-selective electrode for the determination of the antipsychotic ziprasidone in mixed solvents is presented. The electrode contains a plasticized polymeric membrane based on a ziprasidone-tetraphenylborate ion-exchanger. The influence of membrane composition on the electrode response towards ziprasidone in hydroalcoholic solutions was studied. The electrode displayed a stable response in a 2:3 (v/v) methanol/water medium from a ziprasidone concentration of 3 × 10(-6) M with a fast response time of less than 20 s. The electrode also showed good selectivity towards ziprasidone over common inorganic and organic compounds and several species with pharmacological activity. The electrode was successfully applied to the determination of ziprasidone in pharmaceuticals and human urine and serum.

Application of a trazodone-selective electrode to pharmaceutical quality control and urine analyses.

A trazodone-selective electrode for application in pharmaceutical quality control and urine analysis was developed. The electrode is based on incorporation of a trazodone-tetraphenylborate ion exchanger in a poly(vinyl chloride) membrane. The electrode showed a fast, stable and Nernstian response over a wide trazodone concentration range (5 x 10(-5)-1 x 10(-2) M) with a mean slope of 59.3 +/- 0.9 mV/dec of concentration, a mean detection limit of 1.8 x 10(-5) +/- 2.2 x 10(-6) M, a wide working pH range (5-7.5) and a fast response time (less than 20 s). The electrode also showed good accuracy, repeatability, reproducibility and selectivity with respect to some inorganic and organic compounds, including the main trazodone metabolite. The electrode provided good analytical results in the determination of trazodone in pharmaceuticals and spiked urine samples; no extraction steps were necessary. Dissolution testing of trazodone tablets, in different conditions of pH and particle size, based on a direct potentiometric determination with the new selective electrode is presented as well.

Intranasal immunization with a dominant T-cell epitope peptide of a major allergen of olive pollen prevents mice from sensitization to the whole allergen.

Mucosal tolerance induction with vaccines based on peptides representing T-cell epitopes of allergens is a promising way for treating allergic diseases. Ole e 1 is the main allergen of olive pollen, which is an important cause of allergy in Mediterranean countries. The aim of this study was to evaluate the ability of the peptide T109-K130 containing a dominant T-cell epitope of Ole e 1, to modulate the allergen-specific immune response in a prophylactic mouse model. Mice were intranasally treated with the peptide 1 week prior to sensitization with Ole e 1. Blood, lungs and spleens were collected and analysed for immune response. Intranasal pretreatment of mice with the peptide led to suppress serum specific IgE, IgG1 and IgG2a antibody levels, and markedly reduced proliferative T-cell response and Th2-cytokine production, but increased IFN-gamma secretion in spleen cell cultures. Increased mRNA IL-10 levels were observed in lungs from pretreated mice. Pathologic alterations of the lung associated with airway inflammation (peribronchial/perivascular infiltrates, eosinophilia and mucus production) were significantly suppressed after pretreatment. Similar results were obtained when mice were sensitized 10 weeks after treatment. Our results demonstrate that intranasal administration of a single T-cell peptide protects mice against subsequent sensitization to the allergen, possibly via IFN-gamma and IL-10. This study emphasizes the usefulness of nasal peptide T-based vaccines against allergy.

Estudio de prevalencia de la infección por Helicobacter Pylori en una población de Montevideo / Prevalence study of Helicobacter infection in a Montevideo population

El objetivo del trabajo, fue conocer la prevalencia de la infección por HP en una población no seleccionada de Montevideo. Se determinó la presencia de inmunoglobulinas lgG dirigidas contra proteínas de la pared del HP. Se consideraron la edad y el nivel socioeconómico. Se estudiaron 720 sueros, 360 de pacientes de instituciones públicas y 360 de privadas. Se dividieron en 6 grupos etarios, 30 hombres y 30 mujeres cada uno. Se utilizó una técnica inmunoenzimática (Enzignost HP IgG Behring). Hubo seropositividad en 330 individuos (46 por ciento), 55 por ciento en instituciones públicas y 37 por ciento en privadas, con mayor diferencia en los grupos jóvenes (18 por ciento y 2 por ciento). Aumentó de 1O por ciento en menores de 9 años a 63 por ciento en mayores de 60 por ciento. La bibliografía muestra 30 por ciento de infección en países desarrollados y 80 por ciento en subdesarrollados, aumentando con la edad y el medio socioeconómico bajo. La prevalencia en Montevideo es intermedia manteniendo las mismas características