Engineered allosteric ribozymes that respond to specific divalent metal ions

In vitro selection was used to isolate five classes of allosteric hammerhead ribozymes that are triggered by binding to certain divalent metal ion effectors. Each of these ribozyme classes are similarly activated by Mn(2+), Fe(2+), Co(2+), Ni(2+), Zn(2+) and Cd(2+), but their allosteric binding sites reject other divalent metals such as Mg(2+), Ca(2+) and Sr(2+). Through a more comprehensive survey of cations, it was determined that some metal ions (Be(2+), Fe(3+), Al(3+), Ru(2+) and Dy(2+)) are extraordinarily disruptive to the RNA structure and function. Two classes of RNAs examined in greater detail make use of conserved nucleotides within the large internal bulges to form critical structures for allosteric function. One of these classes exhibits a metal-dependent increase in rate constant that indicates a requirement for the binding of two cation effectors. Additional findings suggest that, although complex allosteric functions can be exhibited by small RNAs, larger RNA molecules will probably be required to form binding pockets that are uniquely selective for individual cation effectors

Xenopus paraxis homologue shows novel domains of expression

The paraxis gene encodes a basic helix-loop-helix transcription factor that is expressed in paraxial mesoderm and whose mutant displays an inability to form epithelial somites. Here, the molecular characterization of Xenopus paraxis is reported. paraxis is expressed in the paraxial mesoderm and somites but is down-regulated during muscle differentiation. In addition to its paraxial mesodermal expression, described in other organisms, two novel expression domains of paraxis were found: the neural tube and the head mesoderm, paraxis expression in the neural tube was compared with the expression of the neural markers Xash and Xiro1, and we concluded that paraxis is expressed in a broad band in the prospective sulcus limitans of the neural tube.Fil: Carpio, Ronald. Universidad de Chile; ChileFil: Honore, Stella Maris. Universidad de Chile; Chile. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Instituto Superior de Investigaciones Biológicas. Universidad Nacional de Tucumán. Instituto Superior de Investigaciones Biológicas; ArgentinaFil: Araya, Claudio. Universidad de Chile; ChileFil: Mayor, Roberto. Colegio Universitario de Londres; Reino Unido. Universidad de Chile; Chil

Defining the Critical Hurdles in Cancer Immunotherapy

ABSTRACT: Scientific discoveries that provide strong evidence of antitumor effects in preclinical models often encounter significant delays before being tested in patients with cancer. While some of these delays have a scientific basis, others do not. We need to do better. Innovative strategies need to move into early stage clinical trials as quickly as it is safe, and if successful, these therapies should efficiently obtain regulatory approval and widespread clinical application. In late 2009 and 2010 the Society for Immunotherapy of Cancer (SITC), convened an "Immunotherapy Summit" with representatives from immunotherapy organizations representing Europe, Japan, China and North America to discuss collaborations to improve development and delivery of cancer immunotherapy. One of the concepts raised by SITC and defined as critical by all parties was the need to identify hurdles that impede effective translation of cancer immunotherapy. With consensus on these hurdles, international working groups could be developed to make recommendations vetted by the participating organizations. These recommendations could then be considered by regulatory bodies, governmental and private funding agencies, pharmaceutical companies and academic institutions to facilitate changes necessary to accelerate clinical translation of novel immune-based cancer therapies. The critical hurdles identified by representatives of the collaborating organizations, now organized as the World Immunotherapy Council, are presented and discussed in this report. Some of the identified hurdles impede all investigators, others hinder investigators only in certain regions or institutions or are more relevant to specific types of immunotherapy or first-in-humans studies. Each of these hurdles can significantly delay clinical translation of promising advances in immunotherapy yet be overcome to improve outcomes of patients with cancer