Molecular Tools for Gene Analysis in Fission Yeast

Schizosaccharomyces pombe or fission yeast has been called micromammal due to the potential application of the knowledge derived from the yeast in the physiology of higher eukaryotes. Fission yeast has been consolidated as an excellent model for the study of highly conserved cellular processes. The possibility of using haploid or diploid strains facilitates the analysis of the dominant or recessive phenotype of an allele as well as its function, making it a model of first choice for the development of any investigation in eukaryotes cells. With a growing community that employs fission yeast as a model system for the study of numerous cellular processes, it has motivated the simultaneous development of molecular tools that facilitate the study of genes and proteins in the yeast. In this review, we present the most used molecular techniques in fission yeast for the analysis of genes, its characterization, as well as the determination of its function

Study of Cellular Processes in Higher Eukaryotes Using the Yeast Schizosaccharomyces pombe as a Model

Schizosaccharomyces pombe (Sz. pombe), or fission yeast, is an ascomycete unicellular fungus that has been used as a model system for studying diverse biological processes of higher eukaryotic cells, such as the cell cycle and the maintenance of cell shape, apoptosis, and ageing. Sz. pombe is a rod-shaped cell that grows by apical extension; it divides along the long axis by medial fission and septation. The fission yeast has a doubling time of 2–4 hours, it is easy and inexpensive to grow in simple culture conditions, and can be maintained in the haploid or the diploid state. Sz. pombe can be genetically manipulated using methods such as mutagenesis or gene disruption by homologous recombination. Fission yeast was defined as a micro-mammal because it shares many molecular, genetic, and biochemical features with cells of higher eukaryotes in mRNA splicing, post-translational modifications as N-glycosylation protein, cell-cycle regulation, nutrient-sensing pathways as the target of rapamycin (TOR) network, cAMP-PKA pathway, and autophagy. This chapter uses Sz. pombe as a useful model for studying important cellular processes that support life such as autophagy, apoptosis, and the ageing process. Therefore, the molecular analysis of these processes in fission yeast has the potential to generate new knowledge that could be applied to higher eukaryotes

Assessment of major depressive disorder model in rat

El trastorno depresivo mayor (MDD) es una enfermedad caracterizada por la presencia de dos o más episodios depresivos consecutivos, es un factor de riesgo para cometer suicidio. La mayoría de las terapias antidepresivas se basan en el mantenimiento de la serotonina (5-HT) en la hendidura sináptica. El modelo debulbectomía olfatoria bilateral (OBX) tiene características similares al trastorno depresivo humano. El objetivo de este trabajo es caracterizar a la bulbectomía olfatoria como un modelo de depresión en ratas. Los resultados obtenidos muestran cambios fisiológicos y conductuales que son comparables a la depresión humana, las ratas bulbectomizadas disminuyeron de peso comparadas con los grupos control y cirugía sin extracción, la presencia de la hiperactividad es reducida después de 21 días de tratamiento con antidepresivos, la respuesta alterada a los estímulos adversos y la disminución del acicalamiento puede ser comparado con síntomas de depresión en humanos.Major depressive disorder (MDD) is a disease characterized by the presence of two or more depressive episodes. It is a risk factor to commit suicide. The most common antidepressant therapies are based on maintaining serotonin (5-HT) in the synaptic cleft. The bilateral olfactory bulbectomy (OBX) in rat model have characteristics similar to human depressive disorder. The aim of this work is to characterize olfactory bulbectomy as a depressive major model in rats. These results show physiological and behavioral changes that are comparable to human depression; bulbectomized rats have decreased weight compared to control and surgery without extraction groups, the presence of hyperactivity is reduced after 21 days of treatment with antidepressants (SSRI), altered response to adverse stimuli and a decrease of grooming can be equated a symptom of depression in humans

Swine health: history, challenges and prospects

En los sistemas de producción porcina, uno de los puntos críticos que deben ser atendidos con estricto rigor, es la salud de los cerdos. La salud, es un componente estructural del bienestar animal y refleja un estado óptimo de los animales, lo que repercute directamente en un mayor desempeño productivo y mejores condiciones de desarrollo. Uno de los eslabones más frágiles de la salud de los cerdos, es la presencia de enfermedades infecciosas más importantes, las cuales pueden representar pérdidas hasta del 100 % de la producción, por lo cual, debe ser un tema de atención constante, y continuamente vigilado por el Médico Veterinario Zootecnista y los productores, en perfecta coordinación con las autoridades sanitarias oficiales. En la actualidad, la implementación de mejores prácticas en la cadena productiva es de interés para productores y consumidores. El control de las enfermedades infecciosas debe ser un tema de colaboración entre los diferentes actores del entorno y ser considerado un bien público, ya que las repercusiones negativas, pueden ser desde el nivel local hasta mundial. En la presente revisión, se abordará la temática relacionada con las principales enfermedades infecciosas que ponen en riesgo la salud porcina, el impacto, las principales aportaciones realizadas por el Instituto Nacional de Investigaciones Forestales, Agrícolas y Pecuarias (INIFAP) en sus 35 años de vida, específicamente en el Centro Nacional de Investigación Disciplinaria en Salud Animal e Inocuidad (CENID-SAI), anteriormente conocido como el emblemático CENID-Microbiología o Palo Alto.In swine production systems, one of the critical points that must be strictly attended to is the health of the pigs. Health is a structural component of animal welfare and reflects an optimal state of the animals, which has a direct impact on a higher productive performance and better development conditions. Infectious diseases are one of the greatest threats to the health of pigs and can cause losses of up to 100 % of production; therefore, it requires constant attention and continuous monitoring by the veterinarian and producers, in perfect coordination with the official health authorities. Currently, the implementation of best practices in the production chain is of interest to both producers and consumers. The control of infectious diseases requires collaboration between the various actors in the environment and must be considered a public good, since their negative repercussions can range from the local to the global level. This review will address the main infectious diseases that endanger swine health, their impact, the main contributions made by the National Institute for Research in Forestry, Agriculture and Livestock (INIFAP) in its 35 years of life, mainly at the National Center for Disciplinary Research in Animal Health and Safety (CENID-SAI), formerly known as the emblematic CENID-Microbiología or Palo Alto

Modulation of apoptosis by V protein mumps virus

<p>Abstract</p> <p>Background</p> <p>The Urabe AM9 vaccine strain of mumps virus contains two variants of V protein: VWT (of HN-A1081 viral population) and VGly (of HN-G1081). The V protein is a promoting factor of viral replication by blocking the IFN antiviral pathway.</p> <p>Findings</p> <p>We studied the relationship between V protein variants and IFN-α2b-induced apoptosis. V proteins decrease activation of the extrinsic IFN-α2b-induced apoptotic pathway monitored by the caspase 8 activity, being the effect greater with the VWT protein. Both V proteins decrease the activity of caspase 9 of the intrinsic apoptotic pathway. In a system without IFN, the VWT and VGly proteins expression promotes activation of caspases 3 and 7. However, when the cellular system was stimulated with IFN-α, this activity decreased partially. TUNEL assay shows that for treatment with IFN-α and ibuprofen of cervical adenocarcinoma cells there is nuclear DNA fragmentation but the V protein expression reduces this process.</p> <p>Conclusions</p> <p>The reduction in the levels of caspases and DNA fragmentation, suggesting that V protein, particularly VWT protein of Urabe AM9 vaccine strain, modulates apoptosis. In addition, the VWT protein shows a protective role for cell proliferation in the presence of antiproliferative signals.</p

Interaction of mumps virus V protein variants with STAT1-STAT2 heterodimer: experimental and theoretical studies

<p>Abstract</p> <p>Background</p> <p>Mumps virus V protein has the ability to inhibit the interferon-mediated antiviral response by inducing degradation of STAT proteins. Two virus variants purified from Urabe AM9 mumps virus vaccine differ in their replication and transcription efficiency in cells primed with interferon. Virus susceptibility to IFN was associated with insertion of a non-coded glycine at position 156 in the V protein (VGly) of one virus variant, whereas resistance to IFN was associated with preservation of wild-type phenotype in the V protein (VWT) of the other variant.</p> <p>Results</p> <p>VWT and VGly variants of mumps virus were cloned and sequenced from Urabe AM9 vaccine strain. VGly differs from VWT protein because it possesses an amino acid change Gln₁₀₃Pro (Pro¹⁰³) and the Gly¹⁵⁶insertion. The effect of V protein variants on components of the interferon-stimulated gene factor 3 (ISGF3), STAT1 and STAT2 proteins were experimentally tested in cervical carcinoma cell lines. Expression of VWT protein decreased STAT1 phosphorylation, whereas VGly had no inhibitory effect on either STAT1 or STAT2 phosphorylation. For theoretical analysis of the interaction between V proteins and STAT proteins, 3D structural models of VWT and VGly were predicted by comparing with simian virus 5 (SV5) V protein structure in complex with STAT1-STAT2 heterodimer. <it>In silico </it>analysis showed that VWT-STAT1-STAT2 complex occurs through the V protein Trp-motif (W¹⁷⁴, W¹⁷⁸, W¹⁸⁹) and Glu⁹⁵residue close to the Arg⁴⁰⁹and Lys⁴¹⁵of the nuclear localization signal (NLS) of STAT2, leaving exposed STAT1 Lys residues (K⁸⁵, K⁸⁷, K²⁹⁶, K⁴¹³, K⁵²⁵, K⁶⁷⁹, K⁶⁸⁵), which are susceptible to proteasome degradation. In contrast, the interaction between VGly and STAT1-STAT2 heterodimer occurs in a region far from the NLS of STAT2 without blocking of Lys residues in both STAT1 and STAT2.</p> <p>Conclusions</p> <p>Our results suggest that VWT protein of Urabe AM9 strain of mumps virus may be more efficient than VGly to inactivate both the IFN signaling pathway and antiviral response due to differences in their finest molecular interaction with STAT proteins.</p