Expresión heteróloga de la ATPasa TAA43 y la alcohol deshidrogenasa Ta1316 procedentes de Thermoplasma acidophilum

"El reino Archaea constituye uno de los tres dominios en los que se clasifican los seres vivos. Son microorganismos unicelulares carentes de núcleo y organelos. Fueron descubiertas hace 40 años en ambientes con condiciones extremas y desde entonces se les ha encontrado en todo tipo de hábitats. Debido a que varias especies de arqueas son microorganismos extremófilos poseen propiedades inusuales, lo que los convierte en una valiosa fuente de moléculas útiles en procesos biotecnológicos. Además el patrimonio genético de las arqueas es único ya que poseen genes ortólogos en bacterias y eucariontes, aunado a esto su información genética esta contenida dentro de un solo cromosoma circular pequeño (0.49x106 – 5x106 pares de bases). Todas estas características hacen a las moléculas de los microorganismos del reino Archaea modelos de estudio contra organismos más complejos, además de contribuir (biomasa y biocatalizadores) en el mejoramiento y desarrollo de procesos biotecnológicos. Thermoplasma acidophilum es un microorganismo termo-acidofílico del que actualmente se cuenta con su información genética derivada de la secuenciación de su genoma. En este trabajo se realizó la expresión heteróloga de proteínas provenientes de Thermoplasma acidophilum: 1) la ATPasa TAA43 de la familia de proteínas AAA y 2) las proteínas Ta0841 y Ta1316 ADH que presentan identidad con alcohol deshidrogenasas descritas en las bases de datos existentes. En el caso de la proteína recombinante TAA43 se logró la obtención de condiciones idóneas de cristalización para la elucidación de su estructura tridimensional. Por último, se llevó a cabo la caracterización bioquímica de la alcohol deshidrogenasa Ta1316 ADH, la cual por sus características de termoresistencia se propone como un candidato en los procesos de biocatálisis enzimática.""The AAA proteins (ATPases Associated with various cellular Activities) are enzymes found in all kingdoms of life (Archaea, Bacteria and Eukarya) and they are involved in important cellular functions such as proteolysis and unfolding/folding events to mantain homeostasis. The ORF (Open Reading Frame) Taa43 from Thermoplasma acidophilum genome encodes a 43 kDa protein (TAA43) and contains one AAA domain. TAA43 is classified as a member of the AAA protein family. It remains to be clarified the real function of this protein and the three-dimensional structure would help to clarify its contribution in cellular processes. In this work, the AAA TAA43 protein was expressed in Escherichia coli in an amount of 25 mg/l, a purity of 98% and ATPase activity, with the ultimate purpose of carrying on crystalization assays. We were able to identify growth of crystals conditions for TAA43 and the composition confirmed these preliminary results by X-ray diffraction.

HLA-C genotype and TCR vβ expression analysis in Mexican patients with Psoriasis

"Genetic background and T-cell expansion have been associated as the most important factors for psoriasis susceptibility in the Caucasian population. This study was performed to identify the T cell receptor Vβ repertoire and HLA-Cw genotype in two Mexican groups with severe chronic plaque-type psoriasis. HLA-C typing was performed to detect the allele pattern associated with the disease by sequence-specific primer-polymerase chain reaction. In parallel, RT-PCR and Western blot were used for the identification of the TCR Vβ repertoire. We found a wide variety of HLA-C alleles displayed with a preference to HLA-Cw *07 as the most representative allele in the group of patients. TCR Vβ-2 and Vβ-7 clone-type frequencies were statistically significant (p of 0.0280) when compared to other TCR Vβ expressed in the two groups. We found notable differences both in the HLA-C genotype and TCR Vβ repertoire in the groups of patients studied. Since Mexican individuals are genetically different from the Caucasian population, we suggest that due to these differences the susceptibility to disease and activation of T-cells for a proper immune response may be affected.

Mycobacterium tuberculosis TLR2 agonists LprA, LM and Man-LAM induce notch1 and socs3 transcription

"Mycobacterium tuberculosis employs a number of strategies to subvert host signaling events, leading to its persistence within macrophages. Upon infection, Mycobacterium bovis BCG induce the expression of suppressor of cytokine signaling 3 (socs3), in a Toll-like receptor 2 (TLR2)-Notch1-dependent manner. Purified phosphatidyl inositol di-mannosides (a TLR2 agonist) act as an inducer for the Notch1-socs3 pathway. This prompted us to analyze other TLR2 agonists seeking for additional molecules that may affect this pathway. We found that lipoprotein LprA, as well as glycolipids lipomannan (LM), and mannose-capped lipoarabinomannan (ManLAM) treatment of murine macrophages resulted in stimulation of notch1 and socs3 transcription.

Expresión de receptores notch y detección de virus de papiloma humano en cáncer cervicouterino.

"El desarrollo de cáncer es el resultado de la acumulación progresiva de alteraciones moleculares específicas, particularmente en eventos que dirigen la determinación del destino celular. La vía de señalización Notch juega un papel fundamental en las diferentes etapas de desarrollo celular como proliferación, diferenciación y apoptosis. Estudios recientes han demostrado que dependiendo del nivel de expresión y del contexto celular, los receptores de membrana Notch contribuyen a la tumorogénesis. En este trabajo se investigó el patrón de expresión de los receptores Notch-1 al -4 en cáncer cervicouterino. Se detectó una expresión aumentada de receptores Notch-2 y -3 y una expresión baja pero consistente de Notch-4 en lesiones de pacientes en etapas avanzadas con cáncer cervicouterino. Además se comprobó la presencia de virus de papiloma humano en todos los casos estudiados. Los resultados obtenidos revelan de manera inesperada una expresión diferente a la reportada, ya que el receptor Notch-1 se sobreexpresa y proporciona los efectos para transducir una señal de supervivencia en estas células tumorales.""The development of cancer is the result of a progressive accumulation of specific molecular alterations, mainly those driving cell fate determinations. The Notch signaling pathway plays a crucial role at different stages of cell development such as proliferation, differentiation, and programmed cell death. Recent studies have demonstrated that depending on the expression level and cellular context, the Notch receptors contribute to tumorigenesis. This work describes the expression pattern of the membrane receptors Notch-1 to –4 in cervical cancer. We detected an up-regulated expression of Notch-2 and Notch-3 and a low but steady expression of Notch-4 in lesions of patients with advanced stages of cervical cancer. The presence of the Human Papilloma virus was confirmed in all cases studied. Unexpectedly, the results obtained reveal anatypical expression pattern when compared to the data reported up to now, in which Notch-1 is frequently overexpressed and confers the primary effects to transduce a survival signal in this specific type of tumoral cells.

Vía de señalización Notch y nuevas estrategias para el tratamiento de cáncer Notch signaling pathway and new strategies in cancer treatment

La vía de señalización Notch desempeña un papel fundamental en las diferentes etapas del desarrollo celular como la proliferación, crecimiento, diferenciación y apoptosis. Estudios recientes han demostrado que, dependiendo del nivel de expresión y del contexto celular, los receptores de membrana Notch contribuyen en la resistencia a apoptosis en células tumorales. Estos descubrimientos sugieren que componentes de la vía de señalización Notch son un blanco potencial para el desarrollo de terapias más efectivas contra el cáncer. Esta revisión describe la función de la vía Notch y nuevas estrategias utilizadas en la modulación de su señal.<br>The Notch signaling pathway plays a crucial role at different stages of cell development, such as proliferation, growth, differentiation, and apoptosis. Recent studies demonstrate that depending on the expression level and cellular context, the Notch receptors play a role in apoptosis resistance in malignant cells. These findings suggest that Notch signaling components may be a potential target in the development of new cancer therapies. This review describes the function of the Notch pathway and new strategies in the modulation of its signal

Vía de señalización Notch y nuevas estrategias para el tratamiento de cáncer

La vía de señalización Notch desempeña un papel fundamental en las diferentes etapas del desarrollo celular como la proliferación, crecimiento, diferenciación y apoptosis. Estudios recientes han demostrado que, dependiendo del nivel de expresión y del contexto celular, los receptores de membrana Notch contribuyen en la resistencia a apoptosis en células tumorales. Estos descubrimientos sugieren que componentes de la vía de señalización Notch son un blanco potencial para el desarrollo de terapias más efectivas contra el cáncer. Esta revisión describe la función de la vía Notch y nuevas estrategias utilizadas en la modulación de su señal

Identification and Quantification of Volatile Compounds Found in Vinasses from Two Different Processes of Tequila Production

Vinasses are the main byproducts of ethanol distillation and distilled beverages worldwide and are generated in substantial volumes. Tequila vinasses (TVs) could be used as a feedstock for biohydrogen production through a dark fermentative (DF) process due to their high content of organic matter. However, TV components have not been previously assayed in order to evaluate if they may dark ferment. This work aimed to identify and quantify volatile compounds (VC) in TV and determine if the VC profile depends upon the type of production process (whether the stems were initially cooked or not). TVs were sampled from 3 agave stems with a not-cooking (NC) process, and 3 agave stems with a cooking (C) process, and volatile compounds were determined by gas chromatography coupled with mass spectrometry (GC–MS). A total of 111 volatile compounds were identified, the TV from the cooking process (C) showed the higher presence of furanic compounds (furfural and 5-(hydroxymethyl) furfural) and organic acids (acetic acid and butyric acid), which have been reported as potential inhibitors for DF. To our knowledge, this is the first description of the VC composition from TVs. This study could serve as a base for further investigations related to vinasses from diverse sources

Identification of differential expressed transcripts in cervical cancer of Mexican patients

"The aim of this study was to identify the gene expression profile in biopsies of patients with cervical intraepithelial neoplasia (CIN) 1, CIN 2, CIN 3, and microinvasive cancer by suppression subtractive hybridization and Southern blotting. After analyzing 1,800 cDNA clones, we found 198 upregulated genes, 166 downregulated, and no significant change of gene expression in 86 clones (p = 0.005). These results were validated by Northern blot analysis (p = 0.0001) in the identification of 28 overexpressed and 7 downregulated transcripts. We observed a set of genes related to the Notch signaling pathway that may be involved in the transformation of cervical cells and in the development to malignancy. The differentially expressed genes may provide useful information about the molecular mechanisms involved in human cervical carcinoma and as diagnostic markers.