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The causal agent of brown rot on stone and pome fruits, Monilinia fructicola (G. Wint.), is a quarantine pathogen in Europe. It has been detected in Austria (later eradicated), Spain, the Czech Republic, Italy, Germany, and Switzerland (1). In the United States and other countries, M. fructicola isolates were reported to show resistance to different classes of fungicides, including methyl benzimidazole carbamates (MBC) (2). Lichou et al. (2) reported the presence of isolates resistant to the MBC carbendazim in France, but the mechanisms inducing MBC resistance in these isolates were not studied. Ma et al. (3) in California, and more recently, Zhu et al. (4) in South Carolina, demonstrated that the molecular mechanisms accounting for low and high levels of resistance to MBC fungicides in M. fructicola isolates were the mutations H6Y and E198A, respectively, in the ß-tubulin gene. Four M. fructicola isolates each from Italy, France, Spain, and Switzerland (16 isolates total), all having an unknown level of MBC resistance, were selected. In each isolate, the section of the ß-tubulin gene containing the two potentially mutant codons was PCR-amplified with the primers TubA and TubR1 (3) and the amplicons were sequenced directly. Sequence analysis revealed the amino acid histidine (H) at codon 6 in all the isolates, which would not predict MBC resistance, while alanine (A) at codon 198 (the mutation predictive of a high level of MBC resistance) was found in all isolates from Spain and Switzerland and in three isolates each from France and Italy. A representative sequence of the four identical partial ß-tubulin gene sequences from the Swiss isolates was submitted to GenBank under the Accession No. HQ709265. All isolates were tested in a potato dextrose agar (PDA) petri dish assay for resistance to the MBC fungicide thiophanate-methyl (Nippon Soda Co., Ltd., Tokyo, Japan) at the discriminatory dose of 50 µg/ml (4). All isolates with the E198A mutation were able to grow on the media, while the two isolates without the E198A mutation were not able to grow. The result indicated that most isolates had a high level of resistance to the MBC fungicide. To our knowledge, this is the first report of the presence of the E198A mutation conferring resistance to MBC fungicides in European isolates of M. fructicola. As the mutation appears to be widely distributed, we anticipate that MBC fungicides may be ineffective at controlling brown rot in countries with occurrence of M. fructicola. References: (1) M. Hilber-Bodmer et al. Plant Dis. 94:643, 2010. (2) J. Lichou et al. Phytoma 547:22, 2002. (3) Z. H. Ma et al. Appl. Environ. Microbiol. 69:7145, 2003. (4) F. X. Zhu et al. Plant Dis. 94:1511, 2010.

RESUMEN

We study the control of chaos in an experiment on a parametrically excited pendulum whose excitation mechanism is not perfect. This imperfection leads to a weakly excited degree of freedom with an associated small eigenvalue. Although the state of the pendulum could be characterized well and although the perturbation is weak, we fail to control chaos. From a numerical model we learn that the small eigenvalue cannot be ignored when attempting control. However, the estimate of this eigenvalue from an (experimental) time series is elusive. The reason is that points in an experimental time series are distributed according to the natural measure. It is this extremely uneven distribution of points that thwarts attempts to measure eigenvalues that are very different. Another consequence of the phase-space distribution of points for control is the occurrence of logarithmic-oscillations in the waiting time before control can be attempted. We come to the conclusion that chaos needs to be destroyed before the information needed for its control can be obtained.

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An impact oscillator is a periodically driven system that hits a wall when its amplitude exceeds a critical value. We study impact oscillations where collisions with the wall are with near-zero velocity (grazing impacts). A characteristic feature of grazing impact dynamics is a geometrically converging series of transitions from a nonimpacting period-1 orbit to period-M orbits that impact once per period with M=1,2,ellipsis. In an experiment we explore the dynamics in the vicinity of these period-adding transitions. The experiment is a mechanical impact oscillator with a precisely controlled driving strength. Although the excitation of many high-order harmonics in the experiment appeared unavoidable, we characterize it with only three parameters. Despite the simplicity of this description, good agreement with numerical simulations of an impacting harmonic oscillator was found. Grazing impact dynamics can be described by mappings that have a square-root singularity. We evaluate several mappings, both for instantaneous impacts and for impacts that involve soft collisions with a yielding wall. As the square-root singularity appears persistent in the reduction of the dynamics to mappings, and because impact dynamics appears insensitive to experimental nonidealities, the characteristic bifurcation scenario should be observed in a wide class of experimental systems.

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Hereditary nonpolyposis colorectal carcinoma (HNPCC) is due primarily to inherited mutations in two mismatch repair genes, MSH2 and MLH1, whereas germ-line mutations in other mismatch repair genes are rare. We examined the frequency of germ-line msh6 mutations in a population-based series of 140 colorectal cancer patients, including 45 sporadic cases, 91 familial non-HNPCC cases, and 4 HNPCC cases. Among the 91 population-based familial non-HNPCC cases, germ-line msh6 mutations were found in 6 patients (7.1% of probands analyzed; median age at diagnosis, 61 years). These mutations included a splice site mutation, a frameshift mutation, two missense mutations that were demonstrated to be loss of function mutations, and two missense mutations for which functional studies were not possible. In contrast, germ-line msh6 mutations were not found in any of the 45 sporadic cases and the 4 HNPCC cases in the population-based series or in the second series of 58 clinic-based, primarily HNPCC families. Our data suggest that germ-line msh6 mutations predispose individuals to primarily late-onset, familial colorectal carcinomas that do not fulfill classic criteria for HNPCC.

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The purpose of this study was twofold. In the first part are compared cryosurvival rates for human semen following two different freezing and three different thawing techniques. In the second part a larger number of samples processed by the best of the six methods described were examined to ensure reproducibility. Eighteen human semen samples with initial motility greater than 60% and density greater than 20 X 10(6)/ml were mixed with a cryopreservative medium at 35 degree C and vacuumed into 0.5 ml straws. Six aliquots were prepared from as many specimens as possible. Three straws were frozen by a programmed method (P) and three by a rapid freezing technique (V). All six straws were stored in liquid nitrogen vapor (-196 degree C) for 1 week. The straws were thawed by 1 of 3 methods: (1) room temperature for 10 minutes and 37 degree C hot plate for 10 minutes, (2) ice water for 10 minutes and 37 degree C hot plate for 10 minutes, (3) 35 degree C water for 12 seconds and 37 degree C hot plate for 10 minutes. Postthaw motility was assessed for each aliquot. The freeze/thaw method P/1 was judged optimal. In the second part of the study a larger number of samples were processed by P/1. The mean +/- standard deviation postthaw motility o 57 semen specimens processed by this method was 61.4 +/- 12.1.

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