An NADPH-auxotrophic Corynebacterium glutamicum recombinant strain and used it to construct L-leucine high-yielding strain / Una cepa recombinante NADPH-auxotrófica de Corynebacterium glutamicum y la utilizó para construir una cepa de alto rendimiento de L-leucina

The NADPH-regeneration enzymes in Corynebacterium glutamicum were inactivated to construct an NADPH-auxotrophic C. glutamicum strain by gene knockout and gene replacement. The resultant NADPH-auxotrophic C. glutamicum XL-1 ΔZMICg::ISm (i.e., strain Leu-1) grew well in the basic medium only with gluconate as carbon source. Replacement of the native glyceraldehyde 3-phosphate dehydrogenase (NAD-GapDHCg) by NADP-GapDHCa from Clostridium acetobutylicum is an effective strategy for producing L-leucine in NADPH-prototrophic strain XL-1 and NADPH-auxotrophic strain Leu-1, whereas the L-leucine yield did not differ significantly between these strains (14.1 ± 1.8 g/L vs 16.2 ± 1.1 g/L). Enhancing the carbon flux in biosynthetic pathway by recombinant expression plasmid pEC-ABNCE promoted L-leucine production, but the shortage NADPH supply limited the L-leucine yield. The mutated promoters of zwf and icdCg were introduced into C. glutamicum with NADP-GapDHCa and pEC-ABNCE increased L-leucine yield (54.3 ± 2.9 g/L) and improved cell growth (OD562 = 83.4 ± 7.5) in fed-batch fermentation because the resultant strain C. glutamicum XL-1 ΔMICg::ISm GCg::GCa Pzwf-D1 Picd-D2/pEC-ABNCE (i.e., strain Leu-9) exhibited the proper intracellular NADPH and NADH level. This is the first report of constructing an L-leucine high-yielding strain that reasonably supplies NADPH by optimizing the biosynthetic pathway of NADPH from an NADPH-auxotrophic strain.(AU)

The TRIM proteins in cancer: from expression to emerging regulatory mechanisms

New clinical evidence suggests that dysregulation of the ubiquitin-mediated destruction of tumor suppressors or oncogene products is probably engaged in the etiology of leukemia and carcinoma. The superfamily of tripartite motif (TRIM)-containing protein family is among the biggest recognized single protein RING finger E3 ubiquitin ligases that are considered vital carcinogenesis regulators, which is not shocking since TRIM proteins are engaged in various biological processes, including cell growth, development, and differentiation; hence, TRIM proteins’ alterations may influence apoptosis, cell proliferation, and transcriptional regulation. In this review article, the various mechanisms through which TRIM proteins exert their role in the most prevalent malignancies including lung, prostate, colorectal, liver, breast, brain cancer, and leukemia are summarized.

Microbiota and cancer: current understanding and mechanistic implications

During last few decades, role of microbiota and its importance in several diseases has been a hot topic for research. The microbiota is considered as an accessory organ for maintaining normal physiology of an individual. These microbiota organisms which normally colonize several epithelial surfaces are known to secrete several small molecules leading to local and systemic effects on normal biological processes. The role of microbiota is also established in carcinogenesis as per several recent findings. The effects of microbiota on cancer is not only limited to their contribution in oncogenesis, but the overall susceptibility for oncogenesis and its subsequent progression, development of coinfections, and response to anticancer therapy is also found to be affected by microbiota. The information about microbiota and subsequent contributions of microbes in anticancer response motivated researchers in development of microbes-based anticancer therapeutics. We provided current status of microbiota contribution in oncogenesis with special reference to their mechanistic implications in different aspects of oncogenesis. In addition, the mechanistic implications of bacteria in anticancer therapy are also discussed. We conclude that several mechanisms of microbiota-mediated regulation of oncogenesis is known, but approaches must be focused on understanding contribution of microbiota as a community rather than single organisms-mediated effects.

Emergence of CTNNB1 mutation at acquired resistance to KIT inhibitor in metastatic melanoma

Purpose The KIT inhibitor, imatinib, has shown promising efficacy in patients with KIT-mutated melanoma; however, acquisition of resistance to imatinib occurs rapidly in the majority of patients. The mechanisms of acquired resistance to imatinib in melanoma remain unclear. Methods We analyzed biopsy samples from paired baseline and post-treatment tumor lesions in one patient with KIT-mutated melanoma who had had an initial objective tumor regression in response to imatinib treatment followed by disease progression 8 months later. Results Targeted deep sequencing from post-treatment biopsy samples detected an additional mutation in CTNNB1 (S33C) with original KIT L576P mutation. We examined the functional role of the additional CTNNB1 S33C mutation in resistance to imatinib indirectly using the Ba/F3 cell model. Ba/F3 cell lines transfected with both the L576P KIT mutation and the CTNNB1 S33C mutation demonstrated no growth inhibition despite imatinib treatment, whereas growth inhibition was observed in the Ba/F3 cell line transfected with the L576 KIT mutation alone. Conclusions We report the first identification of the emergence of a CTNNB1 mutation that can confer acquired resistance to imatinib. Further investigation into the causes of acquired resistance to imatinib will be essential to improve the prognosis for patients with KIT mutated melanoma (AU)

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Estudio in vitro sobre los efectos de las estatinas en las curvas de crecimiento celular y su reversión con mevalonato / In vitro study over statins effects on cellular growth curves and its reversibility with mevalonate

Los inhibidores de la HMG-Coa-reductasa, conocidos como estatinas, son los agentes farmacológicos disponibles en el mercado que tienen un mayor efecto hipocolesterolemiante. Los ensayos clínicos y las evidencias experimentales han demostrado que las estatinas tienen un potente efecto antiaterosclerótico. Este efecto es, en parte, la consecuencia del descenso de lípidos, pero también se debe a ciertas acciones pleiotrópicas.Los llamados efectos pleiotrópicos se refieren a distintos aspectos de la función celular: inflamación, coagulación y actividad vasomotora. Estos efectos son mediados, bien indirectamente a través de la reducción del cLDL, o bien por vía directa en las funciones celulares. Aunque muchas de las acciones pleiotrópicas de las estatinas pueden ser un efecto de clase, alguna de ellas puede ser exclusiva de ciertos fármacos y mostrar diferencias en su actividad farmacológica. Así, aunque las estatinas tienen un efecto común sobre los niveles de cLDL, las diferencias en la estructura química y en el perfil farmacocinética pueden motivar variaciones en los efectos pleiotrópicos.En el presente artículo analizamos los efectos in vitro de diferentes estatinas sobre diferentes líneas celulares de células implicadas en el proceso aterogénico: células endoteliales, fibroblastos y células musculares de la pared vascular. En relación con nuestros resultados, comprobamos que los efectos de diferentes dosis de diferentes estatinas sobre las curvas de crecimiento de las diferentes líneas celulares producen diferentes efectos, con independencia de los efectos dependientes de la clase. En consecuencia, los efectos pleiotrópicos sobre el crecimiento celular y su reversibilidad con mevalonato son diferentes según la molécula y la dosis empleadas

HMG-CoA-Reductase inhibitors, also known as statins, are currently the most powerful cholesterol-lowering drugs available on the market. Clinical trials and experimental evidence suggest that statins have heavy anti-atherosclerotic effects. These are in part consequence of lipid lowering but also result from pleiotropic actions of the drugs. These so-called pleiotropic properties affect various aspects of cell function, inflammation, coagulation, and vasomotor activity. These effects are mediated either indirectly through LDL-c reduction or via a direct effect on cellular functions. Although many of the pleiotropic properties of statins may be a class effect, some may be unique to certain agents and account for differences in their pharmacological activity. So, although statins typically have similar effects on LDL-c levels, differences in chemical structure and pharmacokinetic profile can lead to variations in pleiotropic effects. In this paper we analize the in vitro effects of different statins over different cell lines from cells implicated in atherosclerotic process: endothelial cells, fibroblasts, and vascular muscular cells. In relation with our results we can proof that the effects of different dosis of different statins provides singular effects over growth curves of different cellular lines, a despite of a class-dependent effects. So, pleiotropic effects and its reversibility with mevalonate are different according with the molecule and the dosis

Linfedema: métodos de medición y criterios diagnósticos / Lymphedema: Measurement methods and diagnostic criteria

Objetivo: Realizar una revisión y una actualización de los conocimientos existentes respecto a los diferentes métodos de medición del linfedema y los criterios diagnósticos relacionados, cuestiones muy debatidas en las sociedades internacionales de linfología. Estrategia de búsqueda: Se buscaron los artículos originales en las principales bases de datos: Clearinghouse National Guidelines, Guidelines Finder de la National electronic Library for Health del NHS británico, Cochrane Central Register of Controlled Trials, Web of Knowledge y MEDLINE (1996–2009) y las monografías más relevantes sobre linfedema. Selección de estudios: Se valoró la relevancia de los artículos mediante la lectura del título y el resumen, y aquellos considerados relevantes fueron recuperados a texto completo para su análisis posterior. Datos: La medición del tamaño del miembro constituye el aspecto central del diagnóstico y la valoración del linfedema, pudiendo realizarse mediante medidas perimetrales (criometría) o volumetría directa o indirecta. Otras técnicas cuantifican aspectos como las propiedades mecánicas o físicas de los tejidos (AU)

Objective: To review and update the existing knowledge regarding the different methods of measurement of lymphedema and diagnostic criteria, these being very debated issues in international societies of lymphology. Search strategy: Original articles were searched for in major databases: Clearinghouse National Guidelines, Guidelines Finder of the National electronic Library for Health of Britannic NHS, Cochrane Central Register of Controlled Trials, Web of Knowledge and MEDLINE (1996–2009), and the most relevant monographs on Lymphedema. Study selection: The relevance of the articles was evaluated by initial reading of the title and abstract and the full text of those considered relevant was analyzed. Data: The main aspect of the diagnosis and assessment of lymphedema is the measurement of the size of the limb that can be done by circumference measurement or by direct or indirect volumetry. Other methods assess physical or mechanical properties of tissues (AU)

Cytotoxic activity of Alfa-humulene and transcaryophyllene from Salvia officinalis in animal and human tumor cells / Actividad citotóxica del Alfa-humuleno y del tras-cariofileno de Salvia officinalis en dos líneas celulares tumorales animal y humana

Background: The purpose of the present work is two-fold: the fractionationof Salvia officinalis essential oil and the cytotoxic study ofthis oil with its fractions ¡°in vitro¡± tumor cell lines. Materials andMethods: S. officinalis essential oil was obtained by hydrodistillationand fractionated with column chromatography; the essential oil andits fractions were analyzed by gas chromatography (GC) coupled tomass spectrometry (MS). The cytotoxic activity was evaluated incellular lines of breast cancer MCF-7, colon cancer HCT-116, andmurine macrophage RAW264.7 cell lines by the MTT assay. Results:the sub-subfraction F1.1.1 of S. officinalis essential oil containing ¦Á-humulene present highest activity on RAW264.7 and HCT-116 withIC50 values of 41.9 and 77.3 ¦Ìg/ml, respectively. The sub-subfractionF1.2.1 of S. officinalis essential oil with trans-caryophyllene showedless activity on RAW246.7 and HCT-116 with IC50 values of 90.5 and145.8 ¦Ìg/ml. Conclusion: This paper suggests that the ¦Á-humulene andtrans-caryophyllene extracted from S.officinalis essential oil inhibit tumorcell growth (AU)

Antecedentes: Este trabajo tiene dos objetivos: el fraccionamiento del aceite esencial de la especie Salvia officinalis y la determinación de la citotoxicidad del mencionado aceite esencial con sus fracciones en líneas celulares tumorales “in vitro”. Material y Métodos: El aceite esencial de Salvia officinalis fue obtenido por hidrodestilación y fraccionado mediante cromatografía en columna; el aceite esencial y sus fracciones fueron analizadas mediante cromatografía de gases (GC) acoplada a espectrometría de masas (MS). La actividad citotóxica fue evaluada en líneas celulares de cáncer de mama MCF-7; cáncer de colon HCT-116 y en macrófago murino. RAW264.7 con el ensayo MTT. Resultados: La sub-subfracción F1.1.1 del aceite esencial de Salvia officinalis que contiene alfa-humuleno presenta la actividad mas acusada frente a las líneas celulares RAW264.7 y HCT-116, con valores de IC50 de 41,9 y de 77,3 μg/ml respectivamente. La sub-subfracción F1.2.1 del aceite esencial de Salvia officinalis con trans-cariofileno, muestra menor actividad sobre células RAW246.7 y HCT-116 con valores de IC50 de 90,5 y 145,8 μg/ml respectivamente. Conclusión: Estos resultados sugieren que el alfa-humuleno y el trans-cariofileno de los extractos del aceite esencial de Salvia officinalis inhiben el crecimiento de células tumorales (AU)

TSC1-TSC2 complex on the crossroad of pancreatic β cell signaling. Role on cell proliferation, death and survival / Papel del complejo TSC1-TSC2 en la señalización de la célula β pancreática: prolifreración, muerte y supervivencia celular

TSC1-TSC2 complex has emerged as a signal interaction core within the cell. This complex integrates both nutrient and growth factor signaling and is a critical negative regulator of mTORC1.mTORC1 signaling leads to increased protein biosynthesis, which is essential for cell proliferation. Other cellular events such as endoplasmic reticulum stress and autophagy are intimately linked with TSC/mTORC1 pathway and play an important role in pancreatic βcell death or survival. We found that either insulin, glucose independent signaling or the energetic status of the cell are able to modulateTSC2 phosphorylation in pancreatic β cell lines. To show the central role of TSC2 for these cells, we conducted siRNA-mediatedTSC2 silencing. Downregulation of TSC2 leads to an increase inmTORC1/p70S6K signaling, this produces resistance to insulin action. However, specific expression of insulin receptor isoform A restored insulin signalling under these conditions. Moreover, we have explored other processes related to the TSC/mTORC1 pathway andtheir effect on cell death or survival (AU)

El complejo TSC1-TSC2 ha emergido como un núcleo de integración de la señalización de factores de crecimiento y del estado energético celular. Este complejo funciona como un regulador negativo de la actividad de mTORC1. La señalización a través de mTORC1 dirige la síntesis proteica, esencial para la proliferación celular. Otros procesos como el estrés de retículo y la autofagia están íntimamente ligados a la ruta TSC/mTORC1 y juegan un importante papel en la muerte o supervivencia de las células β pancreáticas. Encontramos que tanto la insulina, como la acción independiente de la glucosa o el estatus energético celular modulan la fosforilación de TSC2 en líneas celulares β pancreáticas. Para demostrar el papel central de TSC2 en la regulación de estas células, hemos silenciado su expresión mediante transfección con ARNi. Una menor expresión de TSC2 lleva a un aumento de la señalización demTORC1/p70S6K, lo cual produjo un estado de resistencia a la acción de la insulina. Sin embargo, la expresión selectiva de la isoforma A del receptor de insulina consigue salvar esta resistencia. Por otro lado, hemos explorado otros procesos modulados por la ruta como la autofagiao el estrés de retículo, y su efecto en la supervivencia o muerte celular (AU)