Negative regulation of beta enolase gene transcription in embryonic muscle is dependent upon a zinc finger factor that binds to the G-rich box within the muscle-specific enhancer.

We have previously identified a muscle-specific enhancer within the first intron of the human beta enolase gene. Present in this enhancer are an A/T-rich box that binds MEF-2 protein(s) and a G-rich box (AGTGGGGGAGGGGGCTGCG) that interacts with ubiquitously expressed factors. Both elements are required for tissue-specific expression of the gene in skeletal muscle cells. Here, we report the identification and characterization of a Kruppel-like zinc finger protein, termed beta enolase repressor factor 1, that binds in a sequence-specific manner to the G-rich box and functions as a repressor of the beta enolase gene transcription in transient transfection assays. Using fusion polypeptides of beta enolase repressor factor 1 and the yeast GAL4 DNA-binding domain, we have identified an amino-terminal region responsible for the transcriptional repression activity, whereas a carboxyl-terminal region was shown to contain a potential transcriptional activation domain. The expression of this protein decreases in developing skeletal muscles, correlating with lack of binding activity in nuclear extract from adult skeletal tissue, in which novel binding activities have been detected. These results suggest that in addition to the identified factor, which functionally acts as a negative regulator and is enriched in embryonic muscle, the G-rich box binds other factors, presumably exerting a positive control on transcription. The interplay between factors that repress or activate transcription may constitute a developmentally regulated mechanism that modulates beta enolase gene expression in skeletal muscle

Increase of plasma IL-9 and decrease of plasma IL-5, IL-7, and IFN-γ in patients with chronic heart failure

BACKGROUND: Several cytokines are associated with the development and/or progression of chronic heart failure (CHF). Our aim was to look more closely at the cytokine networks involved in CHF, and to assess whether disease etiology affects cytokine expression. The study population was comprised of a) 69 patients with stable CHF, New York Heart Association (NYHA) II/IV classes, secondary to ischaemic (ICM) and non ischaemic dilated (NIDCM) cardiomyopathy and b) 16 control subjects. We analyzed and compared the plasma levels of 27 pro- and anti-inflammatory mediators, in the study population and assessed for any possible correlation with echocardiographic parameters and disease duration. METHODS: 27 cytokines and growth factors were analyzed in the plasma of ICM- (n = 42) and NIDCM (n = 27) NYHA class II-IV patients vs age- and gender-matched controls (n = 16) by a beadbased multiplex immunoassay. Statistical analysis was performed by ANOVA followed by Tukey post-hoc test for multiple comparison. RESULTS: Macrophage inflammatory protein (MIP)-1\u3b2, Vascular endothelial growth factor (VEGF), interleukin (IL)-9, Monocyte chemotactic protein (MCP)-1, and IL-8 plasma levels were increased in both ICM and NIDCM groups vs controls. In contrast, IL-7, IL-5, and Interferon (IFN)-\u3b3 were decreased in both ICM and NIDCM groups as compared to controls. Plasma IL-6 and IL-1 \u3b2 were increased in ICM and decreased in NIDCM, vs controls, respectively.IL-9 levels inversely correlated, in ICM patients, with left ventricular ejection fraction (LVEF) while IL-5 plasma levels inversely correlated with disease duration, in NYHA III/IV ICM patients.This is the first time that both an increase of plasma IL-9, and a decrease of plasma IL-5, IL-7 and IFN-\u3b3 have been reported in ICM as well as in NIDCM groups, vs controls. Interestingly, such cytokines are part of a network of genes whose expression levels change during chronic heart failure. The altered expression levels of MIP-1 \u3b2, VEGF, MCP-1, IL-1 \u3b2, IL-6, and IL-8, found in this study, are in keeping with previous reports. CONCLUSIONS: The increase of plasma IL-9, and the decrease of plasma IL-5, IL-7 and IFN-\u3b3 in ICM as well as in NIDCM groups vs controls may contribute to get further insights into the inflammatory pathways involved in CHF

Producción de lipasas a partir de hongos endofíticos de Aniba canelilla, caracterización y aplicación del extracto enzimático

Endophytic fungi (EF) have a notable capacity to produce active molecules of industrial importance, such as hydrolytic enzymes. In this study we investigated the production of lipase by EFs isolated from the Amazonian species Aniba canelilla (Lauraceae), characterized the enzymatic extract obtained from the most promising fungus, and applied the lipolytic extract as a biocatalyst in the transesterification reaction for biodiesel production. The fungi were submitted to enzymatic screening in solid medium and in submerged fermentation to assess their lipase production. A total of 292 fungi were tested in solid media. Lipolytic activity was detected in 74% of the fungi cultivated in liquid media, 18 of which showing promising enzymatic production. The best lipase producer, Endomelanconiopsis endophytica QAT_7AC, was identified by sequencing of the ITS region. After adjusting the bioprocess conditions, E. endophytica QAT_7AC produced 2,415.5 U/mL of lipase after 72 h. The enzymatic extract showed higher lipolytic activity under pH 8.0 and 40 oC. The extract was applied as a biocatalyst in a transesterification reaction performed at 40 oC, with ethanol and waste cooking oil (3:1). The biodiesel yield was found to be 87% after 2 h rection when the fungal enzyme was used and 89% with the commercial biocatalyst. The endophytic fungi isolated from A. canelilla proved themselves to be biotechnologically relevant, as they can be explored as potential producers of lipases. The lipolytic extract can be applied in the synthesis of biodiesel using waste cooking oil.Os fungos endofíticos (FE) possuem notável capacidade de produzir moléculas ativas de importância industrial, como as enzimas hidrolíticas. Neste estudo foi investigada a produção de lipase por FEs isolados da espécie amazônica Aniba canelilla (Lauraceae), sendo caracterizado o extrato enzimático obtido a partir do fungo mais promissor e avaliada a aplicação do extrato lipolítico como biocatalisador na reação de transesterificação para a produção de biodiesel. Os fungos foram submetidos à triagem enzimática em meio sólido e por fermentação submersa, para verificar a produção de lipase. Um total de 292 fungos foram testados em meio sólido. A atividade lipolítica detectada em 74% dos fungos cultivados em meio líquido, onde 18 apresentaram atividade enzimática promissora. O melhor produtor de lipase, Endomelanconiopsis endophytica QAT_7AC, foi identificado pelo sequenciamento da região ITS. Após o ajuste das condições do bioprocesso, o E. endophytica QAT_7AC produziu 2.415,5 U/mL de lipase em 72 h. O extrato enzimático apresentou maior atividade lipolítica em pH 8,0 e 40 °C. O extrato foi aplicado como biocatalisador em uma reação de transesterificação realizada a 40 °C, com etanol e óleo de cozinha residual (3:1). O rendimento de biodiesel foi de 87% após 2 h de reação quando a enzima fúngica foi utilizada e de 89% com o biocatalisador comercial. Os fungos endofíticos isolados de A. canelilla mostraram-se biotecnologicamente relevantes e podem ser explorados como potenciais produtores de lipases. O extrato lipolítico pode ser aplicado na síntese de biodiesel a partir do óleo de cozinha residual.Los hongos endofíticos (EF) tienen una notable capacidad de producir moléculas activas de importancia industrial, como las enzimas hidrolíticas. En este estudio investigamos la producción de lipasa por hongos aislados de la especie Amazónica Aniba canelilla (Lauraceae), caracterizamos el extracto enzimático obtenido de lo hongo más promisor y aplicamos el extracto lipolítico como biocatalizador en la reacción de transesterificación para la producción de biodiésel. Los hongos aislados fueron sometidos a un cribado enzimático en medio sólido y fermentación sumergida para avaluar la producción de lipasa. Un total de 292 hongos fueron testados. La actividad lipolítica fue detectada en el 74% de los hongos cultivados en medio líquido, 18 de los cuales presentaron producción enzimática promisoria. El mejor productor de lipasa, Endomelanconiopsis endophytica QAT_7AC, fue identificado por el secuenciamento de la región ITS. Después del ajuste de las condiciones del bioproceso, E. endophytica QAT_7AC produjo 2,415.5 U/mL de lipasa en 72 h. El extracto enzimático presento mayor actividad lipolítica a pH 8.0 y 40 oC. El extracto enzimático fue aplicado como biocatalizador en una reacción de transesterificación realizada a 40 oC con etanol y aceite de cocina usado (3:1). El rendimiento del biodiesel fue de 87% después de 2 h de reacción cuando la enzima fúngica fue utilizada y 89% con el biocatalizador comercial. Los hongos endofíticos aislados de A. canelilla fueron biotecnológicamente relevantes, y pueden ser explorados como potenciales productores de lipasas. El extracto enzimático lipolítico puede ser aplicado en la síntesis de biodiesel utilizando aceite de cocina usado

Actin Cytoskeleton and Regulation of TGFβ Signaling: Exploring Their Links

Human tissues, to maintain their architecture and function, respond to injuries by activating intricate biochemical and physical mechanisms that regulates intercellular communication crucial in maintaining tissue homeostasis. Coordination of the communication occurs through the activity of different actin cytoskeletal regulators, physically connected to extracellular matrix through integrins, generating a platform of biochemical and biomechanical signaling that is deregulated in cancer. Among the major pathways, a controller of cellular functions is the cytokine transforming growth factor β (TGFβ), which remains a complex and central signaling network still to be interpreted and explained in cancer progression. Here, we discuss the link between actin dynamics and TGFβ signaling with the aim of exploring their aberrant interaction in cancer

Actin Cytoskeleton and Regulation of TGFβ Signaling: Exploring Their Links

Human tissues, to maintain their architecture and function, respond to injuries by activating intricate biochemical and physical mechanisms that regulates intercellular communication crucial in maintaining tissue homeostasis. Coordination of the communication occurs through the activity of different actin cytoskeletal regulators, physically connected to extracellular matrix through integrins, generating a platform of biochemical and biomechanical signaling that is deregulated in cancer. Among the major pathways, a controller of cellular functions is the cytokine transforming growth factor β (TGFβ), which remains a complex and central signaling network still to be interpreted and explained in cancer progression. Here, we discuss the link between actin dynamics and TGFβ signaling with the aim of exploring their aberrant interaction in cancer

Transcription factors in fibroblast plasticity and CAF heterogeneity

Abstract In recent years, research focused on the multifaceted landscape and functions of cancer-associated fibroblasts (CAFs) aimed to reveal their heterogeneity and identify commonalities across diverse tumors for more effective therapeutic targeting of pro-tumoral stromal microenvironment. However, a unified functional categorization of CAF subsets remains elusive, posing challenges for the development of targeted CAF therapies in clinical settings. The CAF phenotype arises from a complex interplay of signals within the tumor microenvironment, where transcription factors serve as central mediators of various cellular pathways. Recent advances in single-cell RNA sequencing technology have emphasized the role of transcription factors in the conversion of normal fibroblasts to distinct CAF subtypes across various cancer types. This review provides a comprehensive overview of the specific roles of transcription factor networks in shaping CAF heterogeneity, plasticity, and functionality. Beginning with their influence on fibroblast homeostasis and reprogramming during wound healing and fibrosis, it delves into the emerging insights into transcription factor regulatory networks. Understanding these mechanisms not only enables a more precise characterization of CAF subsets but also sheds light on the early regulatory processes governing CAF heterogeneity and functionality. Ultimately, this knowledge may unveil novel therapeutic targets for cancer treatment, addressing the existing challenges of stromal-targeted therapies

Induction of myogenic differentiation by SDF-1 via CXCR4 and CXCR7 receptors

The stromal cell-derived factor (SDF)-1/CXC receptor 4 (CXCR4) axis has been shown to play a role in skeletal muscle development, but its contribution to postnatal myogenesis and the role of the alternate SDF-1 receptor, CXC receptor 7 (CXCR7), are poorly characterized. Western blot analysis and real-time polymerase chain reaction (PCR) were performed to evaluate in vitro the effect of SDF-1 and CXCR4 and CXCR7 inhibition on myogenic differentiation. Proliferating myoblasts express CXCR4, CXCR7, and SDF-1; during myogenic differentiation, CXCR4 and CXCR7 levels are downregulated, and SDF-1 release is decreased. SDF-1 anticipates myosin heavy chain accumulation and myotube formation in both C2C12 myoblasts and satellite cells. Interestingly, inhibition of CXCR4 and CXCR7 signaling, either by drugs or RNA interfererence, blocks myogenic differentiation. Further, the CXCR4 antagonist, 4F-benzoyl-TN14003, inhibits myoblast cell cycle withdrawal and decreases the retinoblastoma gene (pRb) product accumulation in its hypophosphorylated form. Our experiments demonstrate that SDF-1 regulates myogenic differentiation via both CXCR4 and CXCR7 chemokine receptors. © 2010 Wiley Periodicals, Inc