Active barnase variants with completely random hydrophobic cores.

The central structural feature of natural proteins is a tightly packed and highly ordered hydrophobic core. If some measure of exquisite, native-like core packing is necessary for enzymatic function, this would constitute a significant obstacle to the development of novel enzymes, either by design or by natural or experimental evolution. To test the minimum requirements for a core to provide sufficient structural integrity for enzymatic activity, we have produced mutants of the ribonuclease barnase in which 12 of the 13 core residues have together been randomly replaced by hydrophobic alternatives. Using a sensitive biological screen, we find that a strikingly high proportion of these mutants (23%) retain enzymatic activity in vivo. Further substitution at the 13th core position shows that a similar proportion of completely random hydrophobic cores supports enzyme function. Of the active mutants produced, several have no wild-type core residues. These results imply that hydrophobicity is nearly a sufficient criterion for the construction of a functional core and, in conjunction with previous studies, that refinement of a crudely functional core entails more stringent sequence constraints than does the initial attainment of crude core function. Since attainment of crude function is the critical initial step in evolutionary innovation, the relatively scant requirements contributed by the hydrophobic core would greatly reduce the initial hurdle on the evolutionary pathway to novel enzymes. Similarly, experimental development of novel functional proteins might be simplified by limiting core design to mere specification of hydrophobicity and using iterative mutation-selection to optimize core structure.

A synthetic random basic copolymer with promiscuous binding to class II major histocompatibility complex molecules inhibits T-cell proliferative responses to major and minor histocompatibility antigens in vitro and confers the capacity to prevent murine graft-versus-host disease in vivo.

Graft-versus-host disease (GVHD) is a T-cell-mediated disease of transplanted donor T cells recognizing host alloantigens. Data presented in this report show, to our knowledge, for the first time that a synthetic copolymer of the amino acids L-Glu, L-Lys, L-Ala, and L-Tyr (molecular ratio, 1.9:6.0:4.7:1.0; Mr, 6000-8500) [corrected], termed GLAT, with promiscuous binding to multiple major histocompatibility complex class II alleles is capable of preventing lethal GVHD in the B10.D2 --> BALB/c model (both H-2d) across minor histocompatibility barriers. Administration of GLAT over a limited time after transplant significantly reduced the incidence, onset, and severity of disease. GLAT also improved long-term survival from lethal GVHD: 14/25 (56%) of experimental mice survived > 140 days after transplant compared to 2/26 of saline-treated or to 1/10 of hen egg lysozyme-treated control mice (P < 0.01). Long-term survivors were documented to be fully chimeric by PCR analysis of a polymorphic microsatellite region in the interleukin 1beta gene. In vitro, GLAT inhibited the mixed lymphocyte culture in a dose-dependent fashion across a variety of major barriers tested. Furthermore, GLAT inhibited the response of nylon wool-enriched T cells to syngeneic antigen-presenting cells presenting minor histocompatibility antigens. Prepulsing of the antigen-presenting cells with GLAT reduced the proliferative response, suggesting that GLAT inhibits antigen presentation.

Isolation of virus-neutralizing RNAs from a large pool of random sequences.

RNA and ribonuclease-resistant RNA analogs that bound and neutralized Rous sarcoma virus (RSV) were isolated from a large pool of random sequences by multiple cycles of in vitro selection using infectious viral particles. The selected RNA pool of RSV-binding sequences at a concentration of 0.16 microM completely neutralized the virus. Of 19 sequences cloned from the selected pool, 5 inhibited RSV infection. The selected RNA and RNA analogs were shown to neutralize RSV by interacting with the virus, rather than by adversely affecting the host cells. The selection of the anti-RSV RNA and RNA analogs by intact virions immediately suggests the potential application of this approach to develop RNA and RNA analogs as inhibitors of other viruses such as human immunodeficiency virus.

Mapping sites of interaction of p47-phox and flavocytochrome b with random-sequence peptide phage display libraries.

During assembly of the phagocyte NADPH oxidase, cytosolic p47-phox translocates to the plasma membrane and binds to flavocytochrome b, and binding domains for p47-phox have been identified on the C-terminal tails of both flavocytochrome b subunits. In the present report, we further examine the interaction of these two oxidase components by using random-sequence peptide phage display library analysis. Screening p47-phox with the peptide libraries identified five potential sites of interaction with flavocytochrome b, including three previously reported regions of interaction and two additional regions of interaction of p47-phox with gp91-phox and p22-phox. The additional sites were mapped to a domain on the first predicted cytosolic loop of gp91-phox encompassing residues S86TRVRRQL93 and to a domain near the cytosolic C-terminal tail of gp91-phox encompassing residues F450EWFADLL457. The mapping also confirmed a previously reported binding domain on gp91-phox (E554SGPRGVHFIF564) and putative Src homology 3 domain binding sites on p22-phox (P156PRPP160 and G177GPPGGP183). To demonstrate that the additional regions identified were biologically significant, peptides mimicking the gp91-phox sequences F77LRGSSACCSTRVRRQL93 and E451WFADLLQLLESQ463 were synthesized and assayed for their ability to inhibit NADPH oxidase activity. These peptides had EC50 values of 1 microM and 230 microM, respectively, and inhibited activation when added prior to assembly but did not affect activity of the preassembled oxidase. Our data demonstrate the usefulness of phage display library analysis for the identification of biologically relevant sites of protein-protein interaction and show that the binding of p47-phox to flavocytochrome b involves multiple binding sites along the C-terminal tails of both gp91- and p22-phox and other regions of gp91-phox nearer to the N terminus.

Complete congruence between gene diversity estimates derived from genotypic data at enzyme and random amplified polymorphic DNA loci in black spruce.

Controversy still exists over the adaptive nature of variation of enzyme loci. In conifers, random amplified polymorphic DNAs (RAPDs) represent a class of marker loci that is unlikely to fall within or be strongly linked to coding DNA. We have compared the genetic diversity in natural populations of black spruce [Picea mariana (Mill.) B.S.P.] using genotypic data at allozyme loci and RAPD loci as well as phenotypic data from inferred RAPD fingerprints. The genotypic data for both allozymes and RAPDs were obtained from at least six haploid megagametophytes for each of 75 sexually mature individuals distributed in five populations. Heterozygosities and population fixation indices were in complete agreement between allozyme loci and RAPD loci. In black spruce, it is more likely that the similar levels of variation detected at both enzyme and RAPD loci are due to such evolutionary forces as migration and the mating system, rather than to balancing selection and overdominance. Furthermore, we show that biased estimates of expected heterozygosity and among-population differentiation are obtained when using allele frequencies derived from dominant RAPD phenotypes.

Coherent electron dynamics in a two-dimensional random system with mobility edges

We study numerically the dynamics of a one-electron wavepacket in a two-dimensional random lattice with long-range correlated diagonal disorder in the presence of a uniform electric field. The time-dependent Schrodinger equation is used for this purpose. We find that the wavepacket displays Bloch-like oscillations associated with the appearance of a phase of delocalized states in the strong correlation regime. The amplitude of oscillations directly reflects the bandwidth of the phase and allows us to measure it. The oscillations reveal two main frequencies whose values are determined by the structure of the underlying potential in the vicinity of the wavepacket maximum.

Critical properties of the four-state commutative random permutation glassy Potts model in three and four dimensions

We investigate the critical properties of the four-state commutative random permutation glassy Potts model in three and four dimensions by means of Monte Carlo simulations and a finite-size scaling analysis. By using a field programmable gate array, we have been able to thermalize a large number of samples of systems with large volume. This has allowed us to observe a spin-glass ordered phase in d=4 and to study the critical properties of the transition. In d=3, our results are consistent with the presence of a Kosterlitz-Thouless transition, but also with different scenarios: transient effects due to a value of the lower critical dimension slightly below 3 could be very important.

Phase Transitions in Disordered Systems: The Example of the Random-Field Ising Model in Four Dimensions

By performing a high-statistics simulation of the D = 4 random-field Ising model at zero temperature for different shapes of the random-field distribution, we show that the model is ruled by a single universality class. We compute to a high accuracy the complete set of critical exponents for this class, including the correction-to-scaling exponent. Our results indicate that in four dimensions (i) dimensional reduction as predicted by the perturbative renormalization group does not hold and (ii) three independent critical exponents are needed to describe the transition.

Random exams using Sweave

The adaptation of the Spanish University to the European Higher Education Area (EEES in Spanish) demands the integration of new tools and skills that would make the teaching- learning process easier. This adaptation involves a change in the evaluation methods, which goes from a system where the student was evaluated with a final exam, to a new system where we include a continuous evaluation in which the final exam may represent at most 50% in the vast majority of the Universities. Devising a new and fair continuous evaluation system is not an easy task to do. That would mean a student’s’ learning process follow-up by the teachers, and as a consequence an additional workload on existing staff resources. Traditionally, the continuous evaluation is associated with the daily work of the student and a collection of the different marks partly or entirely based on the work they do during the academic year. Now, small groups of students and an attendance control are important aspects to take into account in order to get an adequate assessment of the students. However, most of the university degrees have groups with more than 70 students, and the attendance control is a complicated task to perform, mostly because it consumes significant amounts of staff time. Another problem found is that the attendance control would encourage not-interested students to be present at class, which might cause some troubles to their classmates. After a two year experience in the development of a continuous assessment in Statistics subjects in Social Science degrees, we think that individual and periodical tasks are the best way to assess results. These tasks or examinations must be done in classroom during regular lessons, so we need an efficient system to put together different and personal questions in order to prevent students from cheating. In this paper we provide an efficient and effective way to elaborate random examination papers by using Sweave, a tool that generates data, graphics and statistical calculus from the software R and shows results in PDF documents created by Latex. In this way, we will be able to design an exam template which could be compiled in order to generate as many PDF documents as it is required, and at the same time, solutions are provided to easily correct them.

Las clases medias en Europa: status y poder en el siglo XXI

European societies are undergoing deep changes as a result of globalization and the international financial crisis that began in 2007. These changes affect the class structure to the extent that these societies suffer an impact on the occupational structure and consume pattern. From the data of Eurobarometer we studied the state of social mobility in the European Union. Comparing the subjective status between 2009 and 2012, we observed divergent movements in European societies. Some societies experimented upward mobility while others just the opposite. We conclude that exists an increasing gap between European societies. To consider the effects of these changes on the status structures a structural model has been tested, exploring the relationship between social class, status and power in European societies. We conclude a statistically significant effect of social class and status on the consciousness perceived to possess more or less power. Two dimensions measure power: “political alienation” and “economic alienation”.

La necesidad de considerar los riesgos climáticos en la introducción de cultivos tropicales en latitudes medias. El mango en el valle del Guadalhorce (Málaga)

El presente trabajo pretende demostrar que no es recomendable la introducción de un cultivo tropical como el mango (Mangifera indica) dentro del área mediterránea del valle del Guadalhorce (Málaga, España). Es un cultivo en expansión por su rentabilidad económica, pero es necesario atender a los posibles riesgos climáticos para su explotación (tras la gran inversión que requiere). Para justificar esta afirmación se presenta un estudio agroclimático realizado en las parcelas experimentales de la finca de IFAPA (Instituto de Investigación y Formación Agraria y Pesquera de Andalucía) de Churriana (Málaga). Los resultados se obtienen a través de los datos de una estación meteorológica y una modelización territorial a partir de herramientas de análisis espacial con SIG. Se tienen en cuenta las variables térmicas y los vientos como condicionantes principales de la aparición de la necrosis apical: patología mortal para el mango.

GSAMPLE: Stata module to draw a random sample

gsample draws a random sample from the data in memory. Simple random sampling (SRS) is supported, as well as unequal probability sampling (UPS), of which sampling with probabilities proportional to size (PPS) is a special case. Both methods, SRS and UPS/PPS, provide sampling with replacement and sampling without replacement. Furthermore, stratified sampling and cluster sampling is supported.