Identifying and Visualizing Functional PAM Diversity across CRISPR-Cas Systems.

CRISPR-Cas adaptive immune systems in prokaryotes boast a diversity of protein families and mechanisms of action, where most systems rely on protospacer-adjacent motifs (PAMs) for DNA target recognition. Here, we developed an in vivo, positive, and tunable screen termed PAM-SCANR (PAM screen achieved by NOT-gate repression) to elucidate functional PAMs as well as an interactive visualization scheme termed the PAM wheel to convey individual PAM sequences and their activities. PAM-SCANR and the PAM wheel identified known functional PAMs while revealing complex sequence-activity landscapes for the Bacillus halodurans I-C (Cascade), Escherichia coli I-E (Cascade), Streptococcus thermophilus II-A CRISPR1 (Cas9), and Francisella novicida V-A (Cpf1) systems. The PAM wheel was also readily applicable to existing high-throughput screens and garnered insights into SpyCas9 and SauCas9 PAM diversity. These tools offer powerful means of elucidating and visualizing functional PAMs toward accelerating our ability to understand and exploit the multitude of CRISPR-Cas systems in nature.

Amyloid fibrils: abnormal protein assembly.

Amyloid refers to the abnormal fibrous, extracellular, proteinaceous deposits found in organs and tissues. Amyloid is insoluble and is structurally dominated by beta-sheet structure. Unlike other fibrous proteins it does not commonly have a structural, supportive or motility role but is associated with the pathology seen in a range of diseases known as the amyloidoses. These diseases include Alzheimer's, the spongiform encephalopathies and type II diabetes, all of which are progressive disorders with associated high morbidity and mortality. Not surprisingly, research into the physicochemical properties of amyloid and its formation is currently intensely pursued. In this chapter we will highlight the key scientific findings and discuss how the stability of amyloid fibrils impacts on bionanotechnology.

Strong isotropic flux pinning in solution-derived YBa2Cu3O7-x nanocomposite superconductor films.

Power applications of superconductors will be tremendously boosted if an effective method for magnetic flux immobilization is discovered. Here, we report the most efficient vortex-pinning mechanism reported so far which, in addition, is based on a low-cost chemical solution deposition technique. A dense array of defects in the superconducting matrix is induced in YBa(2)Cu(3)O(7-x)-BaZrO(3) nanocomposites where BaZrO(3) nanodots are randomly oriented. Non-coherent interfaces are the driving force for generating a new type of nanostructured superconductor. Angle-dependent critical-current measurements demonstrate that a strong and isotropic flux-pinning mechanism is extremely effective at high temperatures and high magnetic fields leading to high-temperature superconductors with record values of pinning force. The maximum vortex-pinning force achieved at 65 K, 78 GN m(-3), is 500% higher than that of the best low-temperature NbTi superconductors at 4.2 K and so a great wealth of high-field applications will be possible at high temperatures.

Biochemical evidence that phaZ gene encodes a specific intracellular medium chain length polyhydroxyalkanoate depolymerase in Pseudomonas putida KT2442: characterization of a paradigmatic enzyme.

Polyhydroxyalkanoates (PHAs) can be catabolized by many microorganisms using intra- or extracellular PHA depolymerases. Most of our current knowledge of these intracellular enzyme-coding genes comes from the analysis of short chain length PHA depolymerases, whereas medium chain length PHA (mcl-PHA) intracellular depolymerization systems still remained to be characterized. The phaZ gene of some Pseudomonas putida strains has been identified only by mutagenesis and complementation techniques as putative intracellular mcl-PHA depolymerase. However, none of their corresponding encoded PhaZ enzymes have been characterized in depth. In this study the PhaZ depolymerase from P. putida KT2442 has been purified and biochemically characterized after its overexpression in Escherichia coli. To facilitate these studies we have developed a new and very sensitive radioactive method for detecting PHA hydrolysis in vitro. We have demonstrated that PhaZ is an intracellular depolymerase that is located in PHA granules and that hydrolyzes specifically mcl-PHAs containing aliphatic and aromatic monomers. The enzyme behaves as a serine hydrolase that is inhibited by phenylmethylsulfonyl fluoride. We have modeled the three-dimensional structure of PhaZ complexed with a 3-hydroxyoctanoate dimer. Using this model, we found that the enzyme appears to be built up from a corealpha/beta hydrolase-type domain capped with a lid structure with an active site containing a catalytic triad buried near the connection between domains. All these data constitute the first biochemical characterization of PhaZ and allow us to propose this enzyme as the paradigmatic representative of intracellular endo/exo-mcl-PHA depolymerases.

Experiencia de la formación especializada en esterilización: una apuesta por la calidad / Specialised training experience in sterilisation: a commitment to queality

La experiencia de la formación especializada en esterilización en el Hospital de Mataró (Consorci Sanitari del Maresme) epezó en el año 2004, cuando desde la Dirección, recibió la directriz de empear a desarrollar un modelo de gestión de calidad para el proceso de esteriización. El modelo elegido fue el ISO 9001 (AU)

Specialised traing experiencie in sterilisation at the Hospital de Matharó (Consorci Sanitari del Maresme) began in 2004, when the Management received guideines on developing a qualitih management model for the sterilisation process. The model selected was the ISO 9001 Standard (AU)