Characterization of the replication of a baculovirus mutant lacking the DNA polymerase gene

AbstractIn a previous study, the DNA polymerase gene (dnapol) of Autographa californica multiple nucleopolyhedrovirus (AcMNPV) was identified as one of six genes required for plasmid replication in a transient replication assay (M. Kool, C. Ahrens, R.W. Goldbach, G.F. Rohrmann, J.M. Vlak, Identification of genes involved in DNA replication of the Autographa californica, Proc. Natl. Acad. Sci. U.S.A. 91, (1994) 11212–11216); however, another study based on a similar approach reported that the virally encoded polymerase was only stimulatory (A. Lu, L.K. Miller, The roles of 18 baculovirus late expression factor genes in transcription and DNA replication, J. Virol. 69, (1995) 975–982). To reconcile the conflicting data and determine if the AcMNPV DNA polymerase is required for viral DNA replication during the course of an infection, a dnapol-null virus was generated using bacmid technology. To detect viral DNA replication, a highly sensitive assay was designed based on real-time PCR and SYBR green chemistry. Our results indicate that a bacmid in which the dnapol ORF was deleted is unable to replicate its DNA when transfected into Spodoptera frugiperda (Sf-9) cells, although when the dnapol ORF was introduced into the polyhedrin (polh) locus, this repaired virus could propagate at levels similar to the control virus. These results confirm that the AcMNPV-encoded DNA polymerase is required for viral DNA replication and the host DNA polymerases cannot substitute for the viral enzyme in this process

Structural and functional analysis of the baculovirus single-stranded DNA-binding protein LEF-3

AbstractThe single-stranded DNA-binding protein LEF-3 of Autographa californica multinucleocapsid nucleopolyhedrovirus consists of 385 amino acid residues, forms oligomers, and promotes Mg2+-independent unwinding of DNA duplexes and annealing of complementary DNA strands. Partial proteolysis revealed that the DNA-binding domain of LEF-3 is located within a central region (residues 28 to 326) that is relatively resistant to proteolysis. In contrast, the N-terminus (27 residues) and C-terminal portion (59 residues) are not involved in interaction with DNA and are readily accessible to proteolytic digestion. Circular dichroism analyses showed that LEF-3 is a folded protein with an estimated α-helix content of more than 40%, but it is structurally unstable and undergoes unfolding in aqueous solutions at temperatures near 50 °C. Unfolding eliminated the LEF-3 domains that are resistant to proteolysis and randomized the digestion pattern by trypsin. The structural transition was irreversible and was accompanied by the generation of high molecular weight (MW) complexes. The thermal treatment inhibited DNA-binding and unwinding activity of LEF-3 but markedly stimulated its annealing activity. We propose that the shift in LEF-3 activities resulted from the generation of the high MW protein complexes, that specifically stimulate the annealing of complementary DNA strands by providing multiple DNA-binding sites and bringing into close proximity the interacting strands. The unfolded LEF-3 was active in a strand exchange reaction suggesting that it could be involved in the production of recombination intermediates

Generalized Dynamic Scaling for Critical Magnetic Systems

The short-time behaviour of the critical dynamics for magnetic systems is investigated with Monte Carlo methods. Without losing the generality, we consider the relaxation process for the two dimensional Ising and Potts model starting from an initial state with very high temperature and arbitrary magnetization. We confirm the generalized scaling form and observe that the critical characteristic functions of the initial magnetization for the Ising and the Potts model are quite different.Comment: 32 pages with15 eps-figure

Radio and optical intra-day variability observations of five blazars

We carried out a pilot campaign of radio and optical band intra-day variability (IDV) observations of five blazars (3C66A, S5 0716+714, OJ287, B0925+504, and BL Lacertae) on December 18--21, 2015 by using the radio telescope in Effelsberg (Germany) and several optical telescopes in Asia, Europe, and America. After calibration, the light curves from both 5 GHz radio band and the optical R band were obtained, although the data were not smoothly sampled over the sampling period of about four days. We tentatively analyse the amplitudes and time scales of the variabilities, and any possible periodicity. The blazars vary significantly in the radio (except 3C66A and BL Lacertae with only marginal variations) and optical bands on intra- and inter-day time scales, and the source B0925+504 exhibits a strong quasi-periodic radio variability. No significant correlation between the radio- and optical-band variability appears in the five sources, which we attribute to the radio IDV being dominated by interstellar scintillation whereas the optical variability comes from the source itself. However, the radio- and optical-band variations appear to be weakly correlated in some sources and should be investigated based on well-sampled data from future observations.Comment: 6 pages, 6 figures, accepted by MNRA

Gr1+IL-4-producing innate cells are induced in response to Th2 stimuli and suppress Th1-dependent antibody responses

Alum is used as a vaccine adjuvant and induces T<sub>h</sub>2 responses and T<sub>h</sub>2-driven antibody isotype production against co-injected antigens. Alum also promotes the appearance in the spleen of Gr1+IL-4+ innate cells that, via IL-4 production, induce MHC II-mediated signaling in B cells. To investigate whether these Gr1+ cells accumulate in the spleen in response to other T<sub>h</sub>2-inducing stimuli and to understand some of their functions, the effects of injection of alum and eggs from the helminth, Schistosoma mansoni, were compared. Like alum, schistosome eggs induced the appearance of Gr1+IL-4+ cells in spleen and promoted MHC II-mediated signaling in B cells. Unlike alum, however, schistosome eggs did not promote CD4 T cell responses against co-injected antigens, suggesting that the effects of alum or schistosome eggs on splenic B cells cannot by themselves explain the T cell adjuvant properties of alum. Accordingly, depletion of IL-4 or Gr1+ cells in alum-injected mice had no effect on the ability of alum to improve expansion of primary CD4 T cells. However, Gr1+ cells and IL-4 played some role in the effects of alum, since depletion of either resulted in antibody responses to antigen that included not only the normal T<sub>h</sub>2-driven isotypes, like IgG1, but also a T<sub>h</sub>1-driven isotype, IgG2c. These data suggest that alum affects the immune response in at least two ways: one, independent of Gr1+ cells and IL-4, that promotes CD4 T cell proliferation and another, via Gr1+IL-4+ cells, that participates in the polarization of the response

Dynamic Simulations of the Kosterlitz-Thouless Phase Transition

Based on the short-time dynamic scaling form, a novel dynamic approach is proposed to tackle numerically the Kosterlitz-Thouless phase transition. Taking the two-dimensional XY model as an example, the exponential divergence of the spatial correlation length, the transition temperature $T_{KT}$ and all critical exponents are computed. Compared with Monte Carlo simulations in equilibrium, we obtain data at temperatures nearer to $T_{KT}$.Comment: to appear in Phys. Rev. E in Rapid Communicatio

Corrections to Scaling for the Two-dimensional Dynamic XY Model

With large-scale Monte Carlo simulations, we confirm that for the two-dimensional XY model, there is a logarithmic correction to scaling in the dynamic relaxation starting from a completely disordered state, while only an inverse power law correction in the case of starting from an ordered state. The dynamic exponent $z$ is $z=2.04(1)$.Comment: to appear as a Rapid commu. in Phys. Rev.

Residual effects of mental fatigue on subjective fatigue, reaction time and cardiac responses

Este estudio investigó los efectos residuales de la fatiga mental inducidos por 30 minutos Test de Stroop con palabra con color incongruente sobre la fatiga percibida subjetivamente, el tiempo de reacción y la variabilidad de la frecuencia cardíaca (HRV) en 20 voluntarios adultos (10 hombres, 10 mujeres). Las variables dependientes se evaluaron antes, inmediatamente después y a los 15, 30, 45 y 60 minutos después de dos condiciones: (a) Test de Stroop con palabra con color incongruente de 30 minutos (condición de fatiga mental); o (b) condición de control de 30 minutos. En la pre condiciones, no hubo diferencias significativas entre las condiciones experimentales para ninguna variable. Sin embargo, hubo un efecto residual de la fatiga mental en las respuestas psicológicas durante hasta ~ 15 minutos después de la manipulación experimental (p < 0,01). Para la tarea de tiempo de reacción, se observaron diferencias significativas solo inmediatamente después de la fatiga mental, en comparación con la condición de control (p < 0,0001). No hubo diferencias significativas entre las condiciones experimentales para ningún parámetro de HRV en ningún período. Además, no hubo diferencias significativas relacionadas con el género en ningún período ni en ninguna de las condiciones para ninguna variable. Los datos sugieren que la fatiga mental puede alterar las respuestas psicológicas hasta ~ 15 min después de su inducción, y la fatiga mental puede afectar el rendimiento del tiempo de reacción cognitiva sin cambiar las respuestas cardíacas autónomas.This study investigated the residual effects of mental fatigue induced by the 30-minute incongruent Stroop Color Word task on subjectively perceived fatigue, reaction time, and heart rate variability (HRV) in 20 adult volunteers (10 men; 10 women). Dependent variables were assessed before, immediately after, and at 15, 30, 45, and 60 minutes after two conditions: (a) a 30-minute incongruent Stroop Color Word task (mental fatigue condition); or (b) a 30-minute control condition. At pre-testing, there were no significant differences between experimental conditions for any variable. However, there was a residual effect of mental fatigue on psychological responses for up to ~15 minutes after the experimental manipulation (p < 0.01). For the reaction time task, significant differences were observed only immediately after the mental fatigue, compared to the control condition (p < 0.0001). There were no significant differences between experimental conditions for any parameters of HRV at any testing period. In addition, there were no significant gender related differences at any period or in either condition for any variable. The data suggest that mental fatigue can alter psychological responses for up to ~15 min after its induction, and mental fatigue may impair cognitive reaction time performance without changing autonomic cardiac responses.Este estudo investigou os efeitos residuais da fadiga mental induzida por 30 minutos do teste de Stroop com a palavra de cor incongruente na fadiga percebida de maneira subjetiva, tempo de reação e variabilidade da frequência cardíaca (VFC) em 20 voluntários adultos (10 homens; 10 mulheres). As variáveis dependentes foram avaliadas antes, imediatamente após e aos 15, 30, 45 e 60 minutos após duas condições: (a) teste de Stroop com a palavra de cor incongruente de 30 minutos (condição fadiga mental); ou (b) uma condição controle de 30 minutos. No pré-teste, não houve diferenças significativas entre as condições experimentais para qualquer variável. No entanto, houve um efeito residual da fadiga mental nas res-postas psicológicas por até ~ 15 minutos após a manipulação experimental (p <0,01). Para o teste de tempo de reação, foram observadas diferenças significativas apenas imediatamente após a fadiga mental, em comparação com a condição controle (p <0,0001). Não houve diferenças significativas entre as condições experimentais para quaisquer parâmetros da VFC em qualquer período. Além disso, não houve diferenças significativas relacionadas ao gênero em qualquer período ou em qualquer condição para qualquer variável. Os dados sugerem que a fadiga mental pode alterar as respostas psico-lógicas por até 15 minutos após sua indução, e a fadiga mental pode prejudicar o desempenho cognitivo pela piora tempo de reação sem alterar as respostas autonômicas cardíacas

Analysis of the effect of locally applied inhomogeneous static magnetic field-exposure on mouse ear edema - a double blind study

The effect static magnetic field (SMF)-exposure may exert on edema development has been investigated. A 6 h long whole-body (WBSMF) or local (LSMF), continuous, inhomogeneous SMF-exposure was applied on anesthetized mice in an in vivo model of mustard oil (MO)-induced ear edema. LSMF was applied below the treated ear, below the lumbar spine, or below the mandible. Ear thickness (v) was checked 8 times during the exposure period (at 0, 0.25, 1, 2, 3, 4, 5, and 6 h). The effect size of the applied treatment (eta) on ear thickness was calculated by the formula eta = 100% x (1-vj/vi), where group i is the control group and j is the treated group. Results showed that MO treatment in itself induced a significant ear edema with an effect of 9% (p11% in both cases compared to SMF-exposure alone (p<0.001). In these cases SMF-exposure alone without MO treatment reduced ear thickness significantly (p<0.05), but within estimated experimental error. In cases of LSMF-exposure on the head, a significant SMF-exposure induced ear thickness reduction was found (eta = 5%, p<0.05). LSMF-exposure on the spine affected ear thickness with and without MO treatment almost identically, which provides evidence that the place of local SMF action may be in the lower spinal region

Short-time dynamics and magnetic critical behavior of two-dimensional random-bond Potts model

The critical behavior in the short-time dynamics for the random-bond Potts ferromagnet in two-dimensions is investigated by short-time dynamic Monte Carlo simulations. The numerical calculations show that this dynamic approach can be applied efficiently to study the scaling characteristic, which is used to estimate the critical exponents theta, beta/nu and z for the quenched disorered systems from the power-law behavior of the kth moments of magnetizations.Comment: 10 pages, 4 figures Soft Condensed Matte