Room-temperature ferromagnetism has been observed in nanoparticles ͑7 -30 nm diam͒ of nonmagnetic oxides such as CeO 2 , Al 2 O 3 , ZnO, In 2 O 3 , and SnO 2 . The saturated magnetic moments in CeO 2 and Al 2 O 3 nanoparticles are comparable to those observed in transition-metal-doped wideband semiconducting oxides. The other oxide nanoparticles show somewhat lower values of magnetization but with a clear hysteretic behavior. Conversely, the bulk samples obtained by sintering the nanoparticles at high temperatures in air or oxygen became diamagnetic. As there were no magnetic impurities present, we assume that the origin of ferromagnetism may be the exchange interactions between localized electron spin moments resulting from oxygen vacancies at the surfaces of nanoparticles. We suggest that ferromagnetism may be a universal characteristic of nanoparticles of metal oxides.Integration of semiconductor with ferromagnetic functionality of electrons has been the focus of recent research in the area of spintronics because of the difficulties associated with the injection of spins into nonmagnetic semiconductors in conventional spintronic devices. Ferromagnetism in semiconductors and insulators is rare, the well-known ferromagnetic semiconductors being the chalcogenides EuX ͑X =O, S, and Se͒ ͑T C Ͻ 70 K͒ and CdCr 2 X 4 ͑X = S and Se͒ ͑T C Ͻ 142 K͒ with the rocksalt and spinel structure, respectively. 1,2 Following the theoretical prediction of Dietl et al. that Mn-doped ZnO and GaN could exhibit ferromagnetism above room temperature, 3 several studies have focused on films and bulk samples of metal oxides such as TiO 2 , ZnO, In 2 O 3 , SnO 2 , and CeO 2 doped with Mn, Co, and other transition metal ions. [4][5][6][7][8] While the existence of ferromagnetism in transitionmetal-doped semiconducting oxides remains controversial, 9 thin films of the band insulator HfO 2 have been reported to exhibit ferromagnetism at room temperature in the absence of any doping. 10 This is puzzling, since pure HfO 2 does not have any magnetic moment and the bulk sample is diamagnetic. Similar ferromagnetism has been reported in other nonmagnetic materials such as CaB 6 , CaO, and SiC where the origin of ferromagnetism is believed to be due to intrinsic defects. 11-13 It has been suggested that ferromagnetism in thin films of HfO 2 may be related to anion vacancies. 14 It has been reported very recently that thin films of undoped TiO 2 and In 2 O 3 also show ferromagnetism at room temperature, 15 the corresponding bulk forms of these materials being diamagnetic. Thin films of these oxides might have defects or oxygen vacancies that could be responsible for the observed ferromagnetism. Ab initio electronic structure calculations using density functional theory in HfO 2 have shown that isolated halfnium vacancies lead to ferromagnetism. 16 Meanwhile, there is a conflicting report attributing the ferromagnetism in HfO 2 to possible iron contamination while using stainless-steel tweezers in handling thin films. 17 In this Rapid Communication, ...

In , FtsLB plays a central role in the initiation of cell division, possibly transducing a signal that will eventually lead to the activation of peptidoglycan remodeling at the forming septum. The molecular mechanisms by which FtsLB operates in the divisome, however, are not understood. Here, we present a structural analysis of the FtsLB complex, performed with biophysical, computational, and methods, that establishes the organization of the transmembrane region and proximal coiled coil of the complex. FRET analysis is consistent with formation of a tetramer composed of two FtsL and two FtsB subunits. We predicted subunit contacts through co-evolutionary analysis and used them to compute a structural model of the complex. The transmembrane region of FtsLB is stabilized by hydrophobic packing and by a complex network of hydrogen bonds. The coiled coil domain probably terminates near the critical constriction control domain, which might correspond to a structural transition. The presence of strongly polar amino acids within the core of the tetrameric coiled coil suggests that the coil may split into two independent FtsQ-binding domains. The helix of FtsB is interrupted between the transmembrane and coiled coil regions by a flexible Gly-rich linker. Conversely, the data suggest that FtsL forms an uninterrupted helix across the two regions and that the integrity of this helix is indispensable for the function of the complex. The FtsL helix is thus a candidate for acting as a potential mechanical connection to communicate conformational changes between periplasmic, membrane, and cytoplasmic regions.

Fluorescence microscopy enables detailed observation of the effects of the antimicrobial peptide Cecropin A on the outer membrane (OM) and cytoplasmic membrane (CM) of single E. coli cells with sub-second time resolution. Fluorescence from periplasmic GFP decays and cell growth halts when the OM is permeabilized. Fluorescence from the DNA stain Sytox Green rises when the CM is permeabilized and the stain enters the cytoplasm. The initial membrane disruptions are localized and stable. Septating cells are attacked earlier than non-septating cells, and curved membrane surfaces are attacked in preference to cylindrical surfaces. Below a threshold bulk Cecropin A concentration, permeabilization is not observed over 30 minutes. Above this threshold, we observe a lag time of several minutes between Cecropin A addition and OM permeabilization and ~30 s between OM and CM permeabilization. The long lag times and the existence of a threshold concentration for permeabilization suggest a nucleation mechanism. However, the roughly linear dependence of mean lag time on bulk peptide concentration is not easily reconciled with a nucleation step involving simultaneous insertion of multiple peptides into the bilayer. Monte Carlo simulations suggest that within seconds the OM permeability becomes comparable to that of a pore of 100-nm diameter, or of numerous small pores distributed over a similarly large area.

Deeper understanding of the bacteriostatic and bactericidal mechanisms of antimicrobial peptides (AMPs) should help in the design of new antibacterial agents. Over several decades, a variety of biochemical assays have been applied to bulk bacterial cultures. While some of these bulk assays provide time resolution on the order of 1 min, they do not capture faster mechanistic events. Nor can they provide subcellular spatial information or discern cell-to-cell heterogeneity within the bacterial population. Single-cell, time-resolved imaging assays bring a completely new spatiotemporal dimension to AMP mechanistic studies. We review recent work that provides new insights into the timing, sequence, and spatial distribution of AMP-induced effects on bacterial cells.

Studies of time-dependent drug and environmental effects on single, live bacterial cells would benefit significantly from a permeable, nonperturbative, long-lived fluorescent stain specific to the nucleoids (chromosomal DNA). The ideal stain would not affect cell growth rate or nucleoid morphology and dynamics, even during laser illumination for hundreds of camera frames. In this study, time-dependent, single-cell fluorescence imaging with laser excitation and a sensitive electron-multiplying chargecoupled-device (EMCCD) camera critically tested the utility of "dead-cell stains" (SYTOX orange and SYTOX green) and "livecell stains" (DRAQ5 and SYTO 61) and also 4=,6-diamidino-2-phenylindole (DAPI). Surprisingly, the dead-cell stains were nearly ideal for imaging live Escherichia coli, while the live-cell stains and DAPI caused nucleoid expansion and, in some cases, cell permeabilization and the halting of growth. SYTOX orange performed well for both the Gram-negative E. coli and the Gram-positive Bacillus subtilis. In an initial application, we used two-color fluorescence imaging to show that the antimicrobial peptide cecropin A destroyed nucleoid-ribosome segregation over 20 min after permeabilization of the E. coli cytoplasmic membrane, reminiscent of the long-term effects of the drug rifampin. In contrast, the human cathelicidin LL-37, while similar to cecropin A in structure, length, charge, and the ability to permeabilize bacterial membranes, had no observable effect on nucleoidribosome segregation. Possible underlying causes are suggested.T he morphology of bacterial nucleoids (the regions containing the chromosomal DNA) is sensitive to the stage of growth (degree of chromosome replication and segregation), to the quality of growth medium, and to the application of external stresses such as nutrient downshift or treatment with drugs (1-5). Highresolution structural studies of bacterial nucleoids using electron microscopy (EM) on slices of fixed cells have a long history (6). Over time, it became clear that nucleoid morphology is highly sensitive to the method of fixation (6, 7). Wide-field fluorescence microscopy lacks the spatial resolution of EM, but it has the important advantage of enabling imaging of live cells as they grow. Live-cell imaging thus offers the possibility of monitoring timedependent changes in nucleoid morphology during normal growth or following application of an external stress.A simple, general method for nonspecific, nonperturbative fluorescent staining of DNA in live bacterial cells remains elusive. One useful strategy places a genetically encoded fluorescent protein on the broadly distributed DNA-binding protein HU (2, 8, 9). However, there is not yet a clear understanding of how HU and other DNA-binding proteins distribute across the chromosomal DNA. As a simple alternative strategy, in this study, we tested a variety of commercially available DNA-staining dyes for their utility in nucleoid imaging for live E. coli cells. When used in flow cytometry assays, these stains are often c...