The Chromosomal Protein Sso7d of the CrenarchaeonSulfolobus solfataricus Rescues Aggregated Proteins in an ATP Hydrolysis-dependent Manner

In this work, we show that the nonspecific DNA-binding protein Sso7d from the crenarchaeon Sulfolobus solfataricus displays a cation-dependent ATPase activity with a pH optimum around neutrality and a temperature optimum of 70 degrees C. Measurements of tryptophan fluorescence and experiments that used 1-anilinonaphthalene-8-sulfonic acid as probe demonstrated that ATP hydrolysis induces a conformational change in the molecule and that the binding of the nucleotide triggers the ATP hydrolysis-induced conformation of the protein to return to the native conformation. We found that Sso7d rescues previously aggregated proteins in an ATP hydrolysis-dependent manner; the native conformation of Sso7d forms a complex with the aggregates, while the ATP hydrolysis-induced conformation is incapable of this interaction. Sso7d is believed to be the first protein isolated from an archaeon capable of rescuing aggregates

Tenencia de pornografía infantil : Penalización y medios probatorios de la comisión del ilícito

El presente trabajo, pretende ahondar en la problemática de la Pornografía Infantil, teniendo presente, que el medio de comisión de tal delito, es el cibernético. Es estas instancias, en donde la tecnología es parte fundamental de la vida de las personas, la pornografía, y en este caso la infantil, han encontrado una nueva manera de ser obtenidas.The present work, intends to delve into the problem of Child Pornography, bearing in mind that the means of commission of such crime, is cybernetic. It is these instances, where technology is a fundamental part of people's lives, pornography, and in this case children's, have found a new way of being obtained.Trabajo final del Seminario Cibercrímenes (JURSOC-UNLP).Facultad de Ciencias Jurídicas y Sociale

Decisiones al final de la vida: Eutanasia : Seminario de Filosofía del Derecho

La muerte nos sitúa en la realidad de lo que constitutivamente somos: seres limitados, finitos, débiles y frágiles. La enfermedad y la muerte son signos evidentes de esta vulnerabilidad. Pero: ¿qué pasa cuando esa enfermedad duele hasta los huesos? ¿Cuándo uno no es más que un recipiente que alberga sólo lamentos físicos, psíquicos y morales? ¿Cómo afrontar la vida, si ya solo queda un respiro? ¿Un tiempo realmente limitado en donde la dignidad humana colapsa y la calidad de vida se ve fuertemente disminuida? La eutanasia, como decisión al final de la vida del individuo, es un tema recurrente, temido, escondido. Hay voces a favor y en contra, pero nadie es neutral: “Acto de compasión”, “Desprecio a la vida humana”; creencias políticas y religiosas que tiñen el panorama y, en el camino, miles de personas padecientes, esperando un milagro que no va a llegar; miles de familias subsumidas en el dolor que conlleva, acompañar a un ser amado en su camino, indigno, hacia la muerte.Death places us in the reality of what we are constitutively: limited, finite, weak and fragile beings. Illness and death are obvious signs of this vulnerability. But what happens when this illness hurts to the bone, when one is no more than a vessel for physical, psychological and moral regrets? How to face life when there is only one breath left, a really limited time in which human dignity collapses and the quality of life is strongly diminished? Euthanasia, as a decision at the end of an individual's life, is a recurrent, feared, hidden topic. There are voices for and against, but no one is neutral: "Act of compassion", "Disregard for human life"; political and religious beliefs that colour the panorama and, along the way, thousands of suffering people, waiting for a miracle that will not come; thousands of families subsumed in the pain that comes with accompanying a loved one on their unworthy path towards death.Facultad de Ciencias Jurídicas y Sociale

Effects of Random Mutagenesis and In Vivo Selection on the Specificity and Stability of a Thermozyme

Factors that give enzymes stability, activity, and substrate recognition result from the combination of few weak molecular interactions, which can be difficult to study through rational protein engineering approaches. We used irrational random mutagenesis and in vivo selection to test if a β-glycosidase from the thermoacidophile Saccharolobus solfataricus (Ssβ-gly) could complement an Escherichia coli strain unable to grow on lactose. The triple mutant of Ssβ-gly (S26L, P171L, and A235V) was more active than the wild type at 85 °C, inactivated at this temperature almost 300-fold quicker, and showed a 2-fold higher kcat on galactosides. The three mutations, which were far from the active site, were analyzed to test their effect at the structural level. Improved activity on galactosides was induced by the mutations. The S26L and P171L mutations destabilized the enzyme through the removal of a hydrogen bond and increased flexibility of the peptide backbone, respectively. However, the flexibility added by S26L mutation improved the activity at T > 60 °C. This study shows that random mutagenesis and biological selection allowed the identification of residues that are critical in determining thermal activity, stability, and substrate recognitio

Criónica, cuando el final de la vida, podría ser la continuación

Ensayo acerca de las implicancias de la criónica y de las practicas transhumanistas, en el derecho y la necesidad de que los legisladores se capaciten en futuras tecnologías para no privar a las personas, en el futuro, de sus derechos fundamentales.Trabajo final para aprobar el Seminario de Bioética y Derecho, dictado por el Dr. Martín Labombarda (JURSOC-UNLP).Facultad de Ciencias Jurídicas y Sociale

THE SYSTEM OF LEXICAL LEVELS IN THE «SYLLABUS OF RUSSIAN LANGUAGE» BY ITALIAN MINISTRY OF EDUCATION FOR THE TEACHERS OF RUSSIAN LANGUAGE IN ITALIAN SECONDARY SCHOOLS

Настоящая статья ставит целью познакомить с важным этапом преподавания русского языка как иностранного в Италии. В 2017 – 2018 гг. под руководством итальянского Министерства образования коллектив итальянских и российских специалистов создал единую программу по обучению РКИ, адресованную пре-подавателям русского языка в итальянских учебных заведениях. Автор вкратце останавливается на критериях разработки лексического материала по уровням и в то же время в соответствии с требованиями русского стандарта по РКИ.=The aim of the article is to introduce to a new important step of the teaching of Russian as a second language in Italy. In 2017 – 2018 the Italian Ministry of Education charged a group of Italian and Russian specialists of Russian as a Foreign Language to create an unitary program, targeted at the teachers of Russian as a second language in Italian sec-ondary schools. The author shortly dwells upon the principles, by which the Russian lexical ma-terials have been organized in connection with the teaching by levels and with the Russian standard Russian as a Foreign Language as well

THE SYSTEM OF LEXICAL LEVELS IN THE «SYLLABUS OF RUSSIAN LANGUAGE» BY ITALIAN MINISTRY OF EDUCATION FOR THE TEACHERS OF RUSSIAN LANGUAGE IN ITALIAN SECONDARY SCHOOLS

Настоящая статья ставит целью познакомить с важным этапом преподавания русского языка как иностранного в Италии. В 2017 – 2018 гг. под руководством итальянского Министерства образования коллектив итальянских и российских специалистов создал единую программу по обучению РКИ, адресованную пре-подавателям русского языка в итальянских учебных заведениях. Автор вкратце останавливается на критериях разработки лексического материала по уровням и в то же время в соответствии с требованиями русского стандарта по РКИ.=The aim of the article is to introduce to a new important step of the teaching of Russian as a second language in Italy. In 2017 – 2018 the Italian Ministry of Education charged a group of Italian and Russian specialists of Russian as a Foreign Language to create an unitary program, targeted at the teachers of Russian as a second language in Italian sec-ondary schools. The author shortly dwells upon the principles, by which the Russian lexical ma-terials have been organized in connection with the teaching by levels and with the Russian standard Russian as a Foreign Language as well

Evaluación de la función respiratória del recien-nacido en el periodo neonatal inmediato

O objetivo do estudo foi Identificar os fatores preditivos e os sinais de taquipnéia transitória neonatal valorizados por enfermeiras de unidade neonatal e alojamento conjunto ao avaliar a função respiratória do recém-nascido no período neonatal imediato. Estudo transversal realizado em duas instituições hospitalares públicas da cidade de São Paulo. Participaram do estudo 28 enfermeiras que responderam um questionário estruturado relacionado às práticas de avaliar a função respiratória do neonato. As enfermeiras consideram relevantes conhecer a evolução do trabalho de parto, parto e o Apgar ao avaliar a função respiratória do neonato. Com relação aos sinais clínicos valorizam o monitoramento da freqüência respiratória, ausculta pulmonar, avaliação do nível de desconforto respiratório utilizando o Índice de Silverman-Andersen, além desses, as enfermeiras de unidade neonatal valorizaram o monitoramento da saturação de oxigênio.The aim of the study was to identify the predictive factors and the signals of transient tachypnea of the newborn valued by the nurses who work at neonatal ward and rooming-in when they assess the newborn respiratory function during the immediate neonatal period in the first six hours after birth. A cross sectional study carried out at two public hospitals of São Paulo city. The sample consisted of 28 nurses who replied a structured questionnaire about the newborn respiratory function assessment practices. Data showed they valuated to know the evaluation of labor, delivery and Apgar score when they evaluate the newborn respiratory function. About the signals of respiratory distress they valued to evaluate the respiratory rate and auscultation, evaluation of the respiratory distress using the Silverman-Andersen Index besides, the neonatal nurses valued to evaluate de oxygen saturation.El objectivo del estudio fue identificar los factores predictivos y los señales de tachypea transitoria del niño valorado por las enfermeras de unidad neonatal y alojamiento conjunto al evaluar la función respiratoria del recién-nascido en el período de neonatal inmediato. Estudio transversal conducido en dos hospitales públicos de la ciudad de São Paulo. Participaran del estúdio 28 enfermeras que responderan un cuestionario estructurado sobre la práctica de evaluar la función respiratorio del niño. Las enfermeras consideran relevante conocer la evolución del trabajo del parto, parto y el escore de Apgar del recién-nacido. Con relación a los señales clinicos, las enfermeras valoraram el monitoramiento de la frecuencia respiratoria, ausculta pulmonar y evaluación de disfunción respiratoria utilizando el Index de Silverman-Andersen. Las enfermeras neonatales también valoraram el monitoramiento de la saturación de oxígeno

Conversion of xylan by recyclable spores of Bacillus subtilis displaying thermophilic enzymes

Background: The Bacillus subtilis spore has long been used to display antigens and enzymes. Spore display can be accomplished by a recombinant and a non-recombinant approach, with the latter proved more efficient than the recombinant one. We used the non-recombinant approach to independently adsorb two thermophilic enzymes, GH10-XA, an endo-1,4-β-xylanase (EC 3.2.1.8) from Alicyclobacillus acidocaldarius, and GH3-XT, a β-xylosidase (EC 3.2.1.37) from Thermotoga thermarum. These enzymes catalyze, respectively, the endohydrolysis of (1-4)-β-d-xylosidic linkages of xylans and the hydrolysis of (1-4)-β-d-xylans to remove successive d-xylose residues from the non-reducing termini. Results: We report that both purified enzymes were independently adsorbed on purified spores of B. subtilis. The adsorption was tight and both enzymes retained part of their specific activity. When spores displaying either GH10- XA or GH3-XT were mixed together, xylan was hydrolysed more efficiently than by a mixture of the two free, not spore-adsorbed, enzymes. The high total activity of the spore-bound enzymes is most likely due to a stabilization of the enzymes that, upon adsorption on the spore, remained active at the reaction conditions for longer than the free enzymes. Spore-adsorbed enzymes, collected after the two-step reaction and incubated with fresh substrate, were still active and able to continue xylan degradation. The recycling of the mixed spore-bound enzymes allowed a strong increase of xylan degradation. Conclusion: Our results indicate that the two-step degradation of xylans can be accomplished by mixing spores displaying either one of two required enzymes. The two-step process occurs more efficiently than with the two un-adsorbed, free enzymes and adsorbed spores can be reused for at least one other reaction round. The efficiency of the process, the reusability of the adsorbed enzymes, and the well documented robustness of spores of B. subtilis indicate the spore as a suitable platform to display enzymes for single as well as multi-step reactions

Prebiotic properties of Bacillus coagulans MA-13: production of galactoside hydrolyzing enzymes and characterization of the transglycosylation properties of a GH42 β-galactosidase

Background: The spore-forming lactic acid bacterium Bacillus coagulans MA-13 has been isolated from canned beans manufacturing and successfully employed for the sustainable production of lactic acid from lignocellulosic biomass. Among lactic acid bacteria, B. coagulans strains are generally recognized as safe (GRAS) for human consumption. Low-cost microbial production of industrially valuable products such as lactic acid and various enzymes devoted to the hydrolysis of oligosaccharides and lactose, is of great importance to the food industry. Specifically, α- and β-galactosidases are attractive for their ability to hydrolyze not-digestible galactosides present in the food matrix as well as in the human gastrointestinal tract. Results: In this work we have explored the potential of B. coagulans MA-13 as a source of metabolites and enzymes to improve the digestibility and the nutritional value of food. A combination of mass spectrometry analysis with conventional biochemical approaches has been employed to unveil the intra- and extra- cellular glycosyl hydrolase (GH) repertoire of B. coagulans MA-13 under diverse growth conditions. The highest enzymatic activity was detected on β-1,4 and α-1,6-glycosidic linkages and the enzymes responsible for these activities were unambiguously identified as β-galactosidase (GH42) and α-galactosidase (GH36), respectively. Whilst the former has been found only in the cytosol, the latter is localized also extracellularly. The export of this enzyme may occur through a not yet identified secretion mechanism, since a typical signal peptide is missing in the α-galactosidase sequence. A full biochemical characterization of the recombinant β-galactosidase has been carried out and the ability of this enzyme to perform homo- and hetero-condensation reactions to produce galacto-oligosaccharides, has been demonstrated. Conclusions: Probiotics which are safe for human use and are capable of producing high levels of both α-galactosidase and β-galactosidase are of great importance to the food industry. In this work we have proven the ability of B. coagulans MA-13 to over-produce these two enzymes thus paving the way for its potential use in treatment of gastrointestinal diseases. [Figure not available: see fulltext.