Expression of Hepatitis C Virus cDNA in Human Hepatoma Cell Line Mediated by a Hybrid Baculovirus–HCV Vector

AbstractAlthough great progress has been made in the characterization of the biochemical and biological features of hepatitis C virus (HCV) gene expression, the elucidation of the HCV life cycle and the evaluation of novel antiviral strategies have been hindered by the lack of a suitable cell culture system. In this context, the development of an efficient HCV cDNA delivery method would contribute to the understanding of HCV replication. To assess the functionality of baculovirus mediated gene delivery for HCV expression, we have constructed recombinant baculoviruses encoding HCV cDNA under the control of the cytomegalovirus promoter. Transduction of the human hepatoma cell line Huh-7 with Bac–HCV vectors was efficient and HCV cDNA expression was enhanced by treatment of the infected cells with dexamethasone. HCV structural and nonstructural polypeptides were processed correctly and were found to localize in the cytoplasm in a pattern characteristic of the endoplasmic reticulum. The expression of the HCV proteins was detected for 49 days after infection. Thus, these results indicate that the recombinant Bac–HCV vectors are a useful tool for the delivery of HCV cDNA and can facilitate the analysis of structural and functional properties of the HCV proteins. In addition, the Bac–HCV vectors can provide important information on the evaluation of novel anti-HCV antiviral strategies

SLN melanoma micrometastasis predictivity of nodal status: a long term retrospective study

BACKGROUND: Completion lymph node dissection (CLND) is the gold standard treatment for patients with a positive sentinel lymph node (SLN) biopsy. Considering the morbidity associated with CLND it is important to identify histological features of the primary tumor and/or of SLN metastasis that could help to spare from CLND a subset of patients who have a very low risk of non-SLN metastasis. The objective of this study is to identify patients with a very low risk to develop non-SLNs recurrences and to limit unnecessary CLND. METHODS: A retrospective long-term study of 80 melanoma patients with positive SLN, undergone CLND, was assessed to define the risk of additional metastasis in the regional nodal basin, on the basis of intranodal distribution of metastatic cells, using the micro-morphometric analysis (Starz classification). RESULTS: This study demonstrates that among the demographic and pathologic features of primary melanoma and of SLN only the Starz classification shows prognostic significance for non-SLN status (p<0.0001). This parameter was also significantly associated with disease-free survival rate (p<0.0013). CONCLUSION: The Starz classification can help to identify, among SLN positive patients, those who can have a real benefit from CLND. From the clinical point of view this easy and reliable method could lead to a significant reduction of unnecessary CLND in association with a substantial decrease in morbidity. The study results indicate that most of S1 subgroup patients might be safely spared from completion lymphatic node dissection. Furthermore, our experience demonstrated that Starz classification of SLN is a safe predictive index for patient stratification and treatment planning

Identification of the most abundant proteins in equine amniotic fluid by a proteomic approach

Characterisation of the physiologic equine amniotic fluid (AF) proteome is a prerequisite to study its changes during diseases and discover new biomarkers. The aim of this study was to identify by a proteomic approach the most abundant proteins of equine AF. AF samples were collected at parturition from 24 healthy mares that delivered healthy foals. All samples were subjected to sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) on 4\u201312% gels. A pool of the 24 samples, after SDS-PAGE, was cut in 25 slices, trypsin-digested and analysed by mass spectrometry (MS) for protein identification. Mean AF protein concentration was 1.96\ua0\ub1\ua01.12\ua0g/L. Thirty-four proteins were successfully identified by MS and subsequently categorised according to Gene Ontology (GO). Twelve proteins (e.g. fibronectin, lumican, thrombospondin and fibulin) belonged to or interacted with the extracellular matrix (ECM) playing an important role in the development of foetal tissues. Most of the remaining proteins were classified as transport (e.g. albumin, major allergen Equ c1 and alpha-fetoprotein) delivering nutrients, ions and lipids essential for foetal growth and development. Among these proteins, major allergen Equ c1 is widely studied in human medicine because it induces Ig-E mediated type I allergic reaction. The absence of immunoglobulins in equine AF was also confirmed

Inhibition of Stearoyl-CoA desaturase 1 reverts BRAF and MEK inhibition-induced selection of cancer stem cells in BRAF-mutated melanoma

Combination therapy with BRAF and MEK inhibitors significantly improves survival in BRAF mutated melanoma patients but is unable to prevent disease recurrence due to the emergence of drug resistance. Cancer stem cells (CSCs) have been involved in these long-term treatment failures. We previously reported in lung cancer that CSCs maintenance is due to altered lipid metabolism and dependent upon Stearoyl-CoA-desaturase (SCD1)-mediated upregulation of YAP and TAZ. On this ground, we investigated the role of SCD1 in melanoma CSCs

Inhibition of Melanogenesis by the Pyridinyl Imidazole Class of Compounds: Possible Involvement of the Wnt/β-Catenin Signaling Pathway

While investigating the role of p38 MAPK in regulating melanogenesis, we found that pyridinyl imidazole inhibitors class compounds as well as the analog compound SB202474, which does not inhibit p38 MAPK, suppressed both α-MSH-induced melanogenesis and spontaneous melanin synthesis. In this study, we demonstrated that the inhibitory activity of the pyridinyl imidazoles correlates with inhibition of the canonical Wnt/β-catenin pathway activity. Imidazole-treated cells showed a reduction in the level of Tcf/Lef target genes involved in the β-catenin signaling network, including ubiquitous genes such as Axin2, Lef1, and Wisp1 as well as cell lineage-restricted genes such as microphthalmia-associated transcription factor and dopachrome tautomerase. Although over-expression of the Wnt signaling pathway effector β-catenin slightly restored the melanogenic program, the lack of complete reversion suggested that the imidazoles interfered with β-catenin-dependent transcriptional activity rather than with β-catenin expression. Accordingly, we did not observe any significant change in β-catenin protein expression. The independence of p38 MAPK activity from the repression of Wnt/β-catenin signaling pathway was confirmed by small interfering RNA knockdown of p38 MAPK expression, which by contrast, stimulated β-catenin-driven gene expression. Our data demonstrate that the small molecule pyridinyl imidazoles possess two distinct and opposite mechanisms that modulate β-catenin dependent transcription: a p38 inhibition-dependent effect that stimulates the Wnt pathway by increasing β-catenin protein expression and an off-target mechanism that inhibits the pathway by repressing β-catenin protein functionality. The p38-independent effect seems to be dominant and, at least in B16-F0 cells, results in a strong block of the Wnt/β-catenin signaling pathway

Role of Wnt canonical pathway in hematological malignancies

Wnt canonical signaling pathway plays a diverse role in embryonic development and maintenance of organs and tissues in adults. It has been observed that Wnt/β-catenin signaling pathway is involved in the pathogenesis of many carcinomas. Moreover, Wnt/β-catenin pathway has been revealed to be associated with angiogenesis. Wnt canonical pathway signaling has great potential as a therapeutic target. It has been disclosed that some hematological malignancies, such as chronic lymphocytic leukemia, mantle cell lymphoma, may occur partly due to the constitutive activation of Wnt canonical signaling pathway. This review will summarize the latest development in Wnt canonical signaling pathway and its roles in tumorigenesis and angiogenesis

Factores que influyen en la elección de las carreras de pedagogía en Colombia

Resumen El presente trabajo tiene como objetivo identificar los factores que influyen sobre las motivaciones, la satisfacción y expectativas en torno al ejercicio profesional en estudiantes de las facultades de Educación y ciclos complementarios de Escuelas Normales Superiores (ENS) en Colombia, tomando caso de estudio la región Caribe de este país. Para ello, se aplicó un estudio de tipo cuantitativo, cuya muestra estuvo integrada por 805 estudiantes de programas afines a los antes mencionados, quienes estuvieron a cargo del diligenciamiento de una encuesta orientada a la medición de los factores que influyen en la elección de estos estudios, de acuerdo con la escala FIT-choice (Factors Influencing Teaching choice), desarrollado por Watt y Richardson (2007). Los resultados obtenidos no solo nos permiten establecer un perfil socioeducativo de los futuros maestros analizados en este trabajo; sino también, facilitan la identificación de los factores vinculados a los principales tipos de motivaciones que tienen mayor peso, al momento de elegir carreras relacionadas con la Pedagogía; siendo las motivaciones de tipo altruistas e intrínsecas las más valoradas por los estudiantes encuestados. Las motivaciones extrínsecas, relacionadas con el salario y la conciliación, por ejemplo, tuvieron una valoración menor, con la excepción del factor relacionado con la seguridad del trabajo. La satisfacción de la elección es elevada pese al bajo salario, la falta de estatus social y una alta disuasión social.This paper aims to identify the factors that influence motivation, satisfaction and expectations about the professional practices among students in education colleges and postsecondary education colleges in Colombia, studying the case of the Caribbean region, based on a quantitative study with 805 students linked with the programs mentioned above, who were in charge of measuring the factors that influence the selection of teaching careers, according to the FIT-choice scale (Factors Influencing Teaching choice), developed by Watt and Richardson (2007). Findings not only allow us to establish a socio-educational profile of future teachers analyzed in this work, but also help us to identify factors linked to the main types of motivations that mostly affect the choice of education careers. In this case, the altruistic and intrinsic motivations are the most valued by respondents, compared with extrinsic motivations, related to salary and time for family, for example, whose valuation was lower. Satisfaction with choice is well valued despite low salaries, lack of social status and high social dissuasion

Lesioni neoplastiche del cavo orale del gatto: aspetti clinici e diagnsotici (studio retrospettivo).

Problema comune nella pratica odontoiatrica veterinaria è determinare precocemente la natura delle lesioni orali e degli aspetti radiografici dell'osso coinvolto. Infatti, sia gli aspetti clinici che radiografici delle lesioni neoplastiche possono mimare patologie benigne. L'obiettivo dello studio è stato quello di valutare gli aspetti clinici e diagnostici in un gruppo di gatti con patologia orale. In 19 su 56 gatti portatori di lesioni dentali e/o orali, è stato avanzato un sospetto di lesione neoplastica. Tuttavia, l'esame isto-patologico ha rivelato che 15 gatti erano realmente affetti da lesione neoplastica, mentre 4 da osteomielite, causata da lesioni da riassorbimento odontoclastico. Anche in questo studio il carcinoma squamocellulare è risultato essere la neoplasi più frequente della cavità orale

A Framework of Major Tumor-Promoting Signal Transduction Pathways Implicated in Melanoma-Fibroblast Dialogue

The development of a modified stromal microenvironment in response to neoplastic onset is a common feature of many tumors including cutaneous melanoma. At all stages, melanoma cells are embedded in a complex tissue composed by extracellular matrix components and several different cell populations. Thus, melanomagenesis is not only driven by malignant melanocytes, but also by the altered communication between melanocytes and non-malignant cell populations, including fibroblasts, endothelial and immune cells. In particular, cancer-associated fibroblasts (CAFs), also referred as melanoma-associated fibroblasts (MAFs) in the case of melanoma, are the most abundant stromal cells and play a significant contextual role in melanoma initiation, progression and metastasis. As a result of dynamic intercellular molecular dialogue between tumor and the stroma, non-neoplastic cells gain specific phenotypes and functions that are pro-tumorigenic. Targeting MAFs is thus considered a promising avenue to improve melanoma therapy. Growing evidence demonstrates that aberrant regulation of oncogenic signaling is not restricted to transformed cells but also occurs in MAFs. However, in some cases, signaling pathways present opposite regulation in melanoma and surrounding area, suggesting that therapeutic strategies need to carefully consider the tumor&ndash;stroma equilibrium. In this novel review, we analyze four major signaling pathways implicated in melanomagenesis, TGF-&beta;, MAPK, Wnt/&beta;-catenin and Hyppo signaling, from the complementary point of view of tumor cells and the microenvironment