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Bacteria growing as surface-adherent biofilms are better able to withstand chemical and physical stresses than their unattached, planktonic counterparts. Using transcriptional profiling and quantitative PCR, we observed a previously uncharacterized gene, yjfO to be upregulated during Escherichia coli MG1655 biofilm growth in a chemostat on serine-limited defined medium. A yjfQ mutant, developed through targeted-insertion mutagenesis, and a yjfQ-complemented strain, were obtained for further characterization. While bacterial surface colonization levels (c.f.u. [cm.sup.-2]) were similar in all three strains, the mutant strain exhibited reduced microcolony formation when observed in flow cells, and greatly enhanced flagellar motility on soft (0.3%) agar. Complementation of yjfO restored microcolony formation and flagellar motility to wild-type levels. Cell surface hydrophobicity and twitching motility were unaffected by the presence or absence of yjfO. In contrast to the parent strain, biofilms from the mutant strain were less able to resist acid and peroxide stresses. yjfO had no significant effect on E. coli biofilm susceptibility to alkali or heat stress. Planktonic cultures from all three strains showed similar responses to these stresses. Regardless of the presence of yjfQ, planktonic E. coli withstood alkali stress better than biofilm populations. Complementation of yjfO restored viability following exposure to peroxide stress, but did not restore acid resistance. Based on its influence on biofilm maturation and stress response, and effects on motility, we propose renaming the uncharacterized gene, yjfO, as bsmA (biofilm stress and motility). DOI 10.1099/mic.0.031468-0

In Drosophila, the enzymatic activity of the glucan binding protein GNBP1 is needed to present Gram-positive peptidoglycan (PG) to peptidoglycan recognition protein SA (PGRP-SA). However, an additional PGRP (PGRP-SD) has been proposed to play a partially redundant role with GNBP1 and PGRP-SA. To reconcile the genetic results with events at the molecular level, we investigated how PGRP-SD participates in the sensing of Gram-positive bacteria. PGRP-SD enhanced the binding of GNBP1 to Gram-positive PG. PGRP-SD interacted with GNBP1 and enhanced the interaction between GNBP1 and PGRP-SA. A complex containing all three proteins could be detected in native gels in the presence of PG. In solution, addition of a highly purified PG fragment induced the occurrence not only of the ternary complex but also of dimeric subcomplexes. These results indicate that the interplay between the binding affinities of different PGRPs provides sufficient flexibility for the recognition of the highly diverse Gram-positive PG. Gram-positive bacteria sensing | pattern recognition receptors

A high-resolution measurement system for the characterization of the delay and angle-of-arrival properties of all significant multipath components on mobile radio channels is presented. The system uses complex impulse response data obtained from a novel 3-D tilted cross switched antenna array as input to an improved version of 3-D Unitary ESPRIT. Results from simulations and measurements show that the approach is capable of accurately characterizing the delay and angle-of-arrival characteristics of the dominant multipath components at moderate urban speeds along a trajectory in mobile propagation environments with high resolution in both azimuth and elevation. Index Terms--Channel sounder, cross arrays, delay estimation, direction of arrival estimation, land mobile radio, mobile communications, radiowave propagation.

In the process of protein synthesis, the small (40S) subunit of the eukaryotic ribosome is recruited to the capped 5' end of the mRNA, from which point it scans along the 5' untranslated region in search of a start codon. However, the 40S subunit alone is not capable of functional association with cellular mRNA species; it has to be prepared for the recruitment and scanning steps by interactions with a group of eukaryotic initiation factors (eIFs). In budding yeast, an important subset of these factors (1, 2, 3, and 5) can form a multifactor complex (MFC). Here, we describe cryo-EM reconstructions of the 40S subunit, of the MFC, and of 40S complexes with MFC factors plus eIF1A. These studies reveal the positioning of the core MFC on the 40S subunit, and show how elF-binding induces mobility in the head and platform and reconfigures the head-platform-body relationship. This is expected to increase the accessibility of the mRNA channel, thus enabling the 40S subunit to convert to a recruitment-competent state. posttranscriptional gene expression | protein synthesis | ribosome structure

This paper concerns the measurement-based comparison of urban microcellular mobile radio channel characteristics at 1.9 GHz and a higher frequency, i.e., 5.8 GHz, where future wireless systems could operate. Characteristics that are reported include transmission loss, root-mean-square delay spread, frequency correlation, and envelope fading statistics. Conclusions are drawn concerning whether significant differences in the performance of mobile radio links operating under the same signal-to-noise ratio conditions in the two bands would be expected in the area where the measurements were made. Index Terms--Correlation bandwidth, fading channels, frequency correlation, frequency-selective fading, multipath channels, radio channel modeling, radio propagation measurements, transmission loss.

The infectious component of hepatitis B (HB) virus (HBV), the Dane particle, has a diameter of [approximately equal to] 44 nm and consists of a double-layered capsid particle enclosing a circular, incomplete double-stranded DNA genome. The outer capsid layer is formed from the HB surface antigen (HBsAg) and lipid, whereas the inner layer is formed from the HB core Ag assembled into an icosahedral structure. During chronic infection HBsAg is expressed in large excess as noninfectious quasispherical particles and tubules with [approximately equal to] 22-nm diameter. Here, we report cryo-EM reconstructions of spherical HBsAg particles at [approximately equal to] 12-[Angstrom] resolution. We show that the particles possess different diameters and have separated them into two predominant populations, both of which have octahedral symmetry. Despite their differing diameters, the two forms of the particle have the same mass and are built through conformational switching of the same building block, a dimer of HBsAg. We propose that this conformational switching, combined with interactions with the underlying core, leads to the formation of HBV Dane particles of different sizes, dictated by the symmetry of the icosahedral core. cryo-EM | subviral particle | virus structure

The structures of membrane-surface bound (prepore) and inserted-pore oligomer forms are determined by using cryo-EM and image processing. These structures provide a direct observation of the conformational transition into the pore form of a cholesterol-dependent cytolysin.

Children's acquisition of English past tense and plural inflection has provided evidence for widely differing theories of morphological representation. The debate has tended to focus on the question of whether [...], In the 'blocking-and-retrieval-failure' account of overregularization (OR; G. F. Marcus, 1995; G. F. Marcus et al., 1992), the claim that a symbolic rule generates regular inflection is founded on pervasively low past tense OR rates and the lack of a substantive difference between past tense and plural OR rates. Evidence of extended periods of OR in the face of substantial correct input (M. Maratsos, 2000) and of an initial period in which nouns are more likely to be overregularized than verbs (V. A. Marchman, K. Plunkett, & J. Goodman, 1997) casts doubt on the blocking account and suggests instead an interplay between type and token frequency effects that is more consistent with usage-based approaches (e.g., J. Bybee, 1995; K. K6pcke, 1998; K. Plunkett & V. Marchman, 1993). However, previous naturalistic studies have been limited by data that account for only 1-2% of child speech. The current study reports analyses of verb and noun ORs in a dense naturalistic corpus (1 child, 2;00.12-3;11.06 [years;months.days])that captures 8-10% of child speech and input. The data show (a) a marked difference in verb and noun OR rates; (b) evidence of a relationship between relative regular/ irregular type frequencies and the onset and rate of past tense and plural ORs; (c) substantial OR periods for some verbs and nouns despite hundreds of correct tokens in child speech and input; and (d) a strong negative correlation between input token frequencies and OR rates for verbs and nouns. The implications of these findings for blocking and other accounts of OR are discussed. KEY WORDS: language acquisition, morphology, overregularization, past tense, plural

Colorectal cancer is one of the most significant causes of cancer death. A genetic model for colorectal cancer has been proposed in which the sequential accumulation of mutations in specific genes, including adenomatous polyposis coli (APC), Kirsten-ras (K-ras), and p53, drives the transition from healthy colonic epithelia through increasingly dysplastic adenoma to colorectal cancer. We have characterized tumor mutation spectra in a large cohort of colorectal cancer patients. In marked contrast to the predictions of the sequential model of mutation accumulation, only 6.6% of tumors were found to contain mutations in APC, K-ras, and p53, with 38.7% of tumors containing mutations in only one of these genes. The most common combination of mutations was p53 and APC (27.1%), whereas mutations in both p53 and K-ras were extremely rare. Statistical analysis (two-sided Fisher's exact test) confirmed that mutations in K-ras and p53 co-occurred less frequently than expected by chance (P < 0.01, Fisher's exact test). This finding suggests that these mutations lie on alternate pathways of colorectal tumor development. The heterogeneous pattern of tumor mutations in our patient cohort suggests that multiple alternative genetic pathways to colorectal cancer exist and that the widely accepted genetic model of cancer development is not representative of the majority of colorectal tumors.

A study was conducted to determine the structure of soluble pneumolysin monomers that are produced by the human pathogen Streptococcus pneumoniae. Using cryo-electron microscopy, the three-dimensional structure of a helical oligomer of pneumolysin and of a membrane-bound ring form was revealed. The diameter of the helical form was estimated to be at 41 subunits per turn while a number of subunits per ring was observed to be in the range of 40-50.

More than 40 calcite-precipitation experiments were performed under sterile conditions in order to investigate the origins of 25-300 nm spherical-, rod-, and ovoid-shaped objects that have been widely interpreted as evidence of nanometer-scale life (i.e., nannobacteria). Individual experiments included the addition of soluble organic compounds, common species of eubacteria, or phage-induced eubacterial lysates. These experiments indicate that many of the nanometer-scale objects have inorganic or nonnannobacterial origins. In the precipitation experiments, calcite formed euhedral crystals 50-800 nm in diameter and smaller (< 50 nm) anhedral or rounded particles or protocrystals. The small anhedral or rounded solids resembled nannobacteria. The relative amount of anhedral or rounded calcite was greatest in experiments with a dissolved organic component. These controlled experiments are in accord with observations that rounded nanometerscale objects are more common in minerals formed in organic-rich environments. Bacterial fragments occur as rounded to irregularly shaped particles that included cell-wall fragments, expulsed cytoplasm, and relict capsules that also closely resembled nannobacteria. Acid etching of the large euhedral crystals produced in the precipitation experiments also resulted in the formation of nanometer-scale features that resembled nannobacteria in natural carbonates. The shapes of the etching artifacts vary as a function of the strength of the acid and the duration of etching. Much caution is advisable in interpreting the origin of rounded features < 50 nm.