Phosphatidylglycerol Is the Lipid Donor for Synthesis of Phospholipid-Linked Enterobacterial Common Antigen.

The Gram-negative outer membrane (OM) is an asymmetric bilayer with phospholipids in its inner leaflet and mainly lipopolysaccharide (LPS) in its outer leaflet and is largely impermeable to many antibiotics. In Enterobacterales (e.g., Escherichia, Salmonella, Klebsiella, and Yersinia), the outer leaflet of the OM also contains phosphoglyceride-linked enterobacterial common antigen (ECAPG). This molecule consists of the conserved ECA carbohydrate linked to diacylglycerol-phosphate (DAG-P) through a phosphodiester bond. ECAPG contributes to the OM permeability barrier and modeling suggests that it may alter the packing of LPS molecules in the OM. Here, we investigate, in Escherichia coli K-12, the reaction synthesizing ECAPG from ECA precursor linked to an isoprenoid carrier to identify the lipid donor that provides the DAG-P moiety to ECAPG. Through overexpression of phospholipid biosynthesis genes, we observed alterations expected to increase levels of phosphatidylglycerol (PG) increased the synthesis of ECAPG, whereas alterations expected to decrease levels of PG decreased the synthesis of ECAPG. We discovered depletion of PG levels in strains that could synthesize ECAPG, but not other forms of ECA, causes additional growth defects, likely due to the buildup of ECA precursor on the isoprenoid carrier inhibiting peptidoglycan biosynthesis. Our results demonstrate ECAPG can be synthesized in the absence of the other major phospholipids (phosphatidylethanolamine and cardiolipin). Overall, these results conclusively demonstrate PG is the lipid donor for the synthesis of ECAPG and provide a key insight into the reaction producing ECAPG. In addition, these results provide an interesting parallel to lipoprotein acylation, which also uses PG as its DAG donor. IMPORTANCE The Gram-negative outer membrane is a permeability barrier preventing cellular entry of antibiotics. However, outer membrane biogenesis pathways are targets for small molecule development. Here, we investigate the synthesis of a form of enterobacterial common antigen (ECA), ECAPG, found in the outer membrane of Enterobacterales (e.g., Escherichia, Salmonella, and Klebsiella). ECAPG consists of the conserved ECA carbohydrate unit linked to diacylglycerol-phosphate-ECA is a phospholipid headgroup. The details of the reaction forming this molecule from polymerized ECA precursor are unknown. We determined the lipid donor providing the phospholipid moiety is phosphatidylglycerol. Understanding the synthesis of outer membrane constituents such as ECAPG provides the opportunity for development of molecules to increase outer membrane permeability, expanding the antibiotics available to treat Gram-negative infections.

"It Depends on Where You Are and What Job You Do": Differences in Tobacco Use across Career Fields in the United States Air Force.

While tobacco use within the military is often discussed as being homogenously part of U.S. military culture, literature from civilian populations highlights that tobacco use varies by career field (e.g., "white collar" vs. "blue collar"). The objective of this qualitative study was to compare tobacco use by career fields in the U.S. Air Force. Airmen, Military Training Instructors, and Technical Training Instructors participated in 22 focus groups across five major Air Force Technical Training bases. Focus groups were conducted in-person using semi-structured interview guides and were audio-recorded. A conventional content-coding approach was used to code transcripts. Participants described substantial variation across the careers, which was attributed to social norms and the nature of jobs. Individuals in careers that spend most of their time outside were more likely to permit tobacco use. Conversely, tobacco use was seen as stigmatized in medical fields. Additionally, smokeless tobacco was identified as popular in certain careers because it could be used covertly on the job. Findings suggest that a one-size-fits-all approach to reducing tobacco use through policies and programs may not reflect the realities of military tobacco use. These findings may provide insights into other branches of the U.S. military with similar career fields.

Plasmonic colors in titanium nitride for robust and covert security features.

A mechanically robust metasurface exhibiting plasmonic colors across the visible and the near-IR spectrum is designed, fabricated, and characterized. Thin TiN layers (41 nm in thickness) prepared by plasma-enhanced atomic layer deposition (ALD) are patterned with sub-wavelength apertures (75 nm to 150 nm radii), arranged with hexagonal periodicity. These patterned films exhibit extraordinary transmission in the visible and the near-IR spectrum (550 nm to 1040 nm), which is accessible by conventional Si CCD detectors. The TiN structures are shown to withstand high levels of mechanical stresses, tested by rubbing the films against a lint-free cloth under 14.5 kPa of load for 30 minutes, while structures patterned on gold, a widely used plasmonic material, do not. The subwavelength nature of the plasmonic resonances, coupled with robustness and durability of TiN, makes these structures an attractive choice for use in nanoscale security features for heavily handled objects. Furthermore, ALD of these films enables scalability, which in conjunction with the cost-effectiveness of the process and material (TiN) makes the entire process industry friendly.

Corrigendum: Controlling hybrid nonlinearities in transparent conducting oxides via two-colour excitation.

This corrects the article DOI: 10.1038/ncomms15829.

Controlling hybrid nonlinearities in transparent conducting oxides via two-colour excitation.

Nanophotonics and metamaterials have revolutionized the way we think about optical space (ɛ,µ), enabling us to engineer the refractive index almost at will, to confine light to the smallest of the volumes, and to manipulate optical signals with extremely small footprints and energy requirements. Significant efforts are now devoted to finding suitable materials and strategies for the dynamic control of the optical properties. Transparent conductive oxides exhibit large ultrafast nonlinearities under both interband and intraband excitations. Here we show that combining these two effects in aluminium-doped zinc oxide via a two-colour laser field discloses new material functionalities. Owing to the independence of the two nonlinearities, the ultrafast temporal dynamics of the material permittivity can be designed by acting on the amplitude and delay of the two fields. We demonstrate the potential applications of this novel degree of freedom by dynamically addressing the modulation bandwidth and optical spectral tuning of a probe optical pulse.

Enhanced Nonlinear Refractive Index in ε-Near-Zero Materials.

New propagation regimes for light arise from the ability to tune the dielectric permittivity to extremely low values. Here, we demonstrate a universal approach based on the low linear permittivity values attained in the ε-near-zero (ENZ) regime for enhancing the nonlinear refractive index, which enables remarkable light-induced changes of the material properties. Experiments performed on Al-doped ZnO (AZO) thin films show a sixfold increase of the Kerr nonlinear refractive index (n_{2}) at the ENZ wavelength, located in the 1300 nm region. This in turn leads to ultrafast light-induced refractive index changes of the order of unity, thus representing a new paradigm for nonlinear optics.

Experimental demonstration of titanium nitride plasmonic interconnects.

An insulator-metal-insulator plasmonic interconnect using TiN, a CMOS-compatible material, is proposed and investigated experimentally at the telecommunication wavelength of 1.55 µm. The TiN waveguide was shown to obtain propagation losses less than 0.8 dB/mm with a mode size of 9.8 µm on sapphire, which agree well with theoretical predictions. A theoretical analysis of a solid-state structure using Si(3)N(4) superstrates and ultra-thin metal strips shows that propagation losses less than 0.3 dB/mm with a mode size of 9 µm are attainable. This work illustrates the potential of TiN as a realistic plasmonic material for practical solid-state, integrated nano-optic and hybrid photonic devices.

Antigenic variation of SIV: mutations in V4 alter the neutralization profile.

Antigenic variation is a characteristic feature of lentiviral infection. The SIV/macaque model of AIDS provides an ideal system in which to investigate the molecular basis of antigenic variation. The purpose of this study was to genetically map the nucleotide changes in env that alter the neutralization phenotype of SIV. Serum taken from an SIVmac239-infected macaque (2D) at 30 weeks postinoculation was found to neutralize the input virus (SIVmac239) and an isolate, P9, obtained at 10 weeks p.i., but did not neutralize two other isolates, P13 and P23, obtained at 20 and 52 weeks, respectively. Sequence analysis of these virus variants revealed clustered amino acid changes in V1 and single base pair changes in V2-V4 of P13 and P23. Infectious recombinant viruses in which the V1 and V1-V3 sequences of SIVmac239 were replaced with those of P13 or P23 retained the neutralization profile of SIVmac239; both were neutralized by macaque 2D serum. Recombinants containing the entire surface glycoprotein (gp120) (V1-V5) and the 5' portion of gp41 of P13 and P23 and those containing gp120 sequences from V4 through the 5' portion of the transmembrane glycoprotein (gp41) were not neutralized by 2D serum. Using a panel of monoclonal antibodies in radioimmunoprecipitation assays, P23 and recombinants containing V4 and V5 of P23 were shown to be antigenically distinct from P13 and SIVmac239. The majority of the amino acid changes in the antigenically distinct viruses were clustered in V4 (amino acids 413-418) and these changes created new potential N-linked glycosylation sites. This study demonstrates that a small number of specific amino acid changes (amino acids 412 to 418 in the env gene) in the V4 region of the SIV envelope glycoprotein can alter antibody recognition and neutralization and that these phenotypic changes may be associated with altered glycosylation of the envelope.

Nucleotide sequence of a 13.9 kb segment of the 90 kb virulence plasmid of Salmonella typhimurium: the presence of fimbrial biosynthetic genes.

The 90kb plasmid resident in Salmonella typhimurium confers increased virulence in mice by promoting the spread of infection after invasion of the intestinal epithelium. The nucleotide sequence of a 13.9kb segment of this plasmid known to encode an outer membrane protein related in sequence to components of fimbrial biosynthesis in enteric bacteria was determined. This cloned segment between the repB and repC replicon regions programmed expression of abundant surface fimbriae in Escherichia coli and S. typhimurium cells. A 7kb region contained seven open reading frames, the protein products of five of which were related in sequence to regulatory, structural, and assembly proteins of adherence fimbriae/pili, such as the P and K88 pili. These five genes and two adjacent ones which were not markedly related to proteins in the data bases comprise the pef (plasmid-encoded fimbriae) locus. Transposon TnphoA insertions in four genes in the pef locus (pefA, pefC, orf5 and orf6) resulted in active PhoA fusions and blocked or reduced the surface presentation of fimbriae, indicating that the proteins encoded by these four genes are translocated at least across the cytoplasmic membrane and contribute to formation of the fimbrial structure. The differences in genetic organization and protein sequence relatedness from other fimbrial gene clusters suggest that the pef locus might encode a novel type of fimbria. Between the pef and the repB loci, there were five open reading frames, one of which (orf8) gave rise to active PhoA fusions but was not necessary for fimbrial expression. Two of the other proteins were homologous to transcription regulatory proteins and a third was the rck gene, which encodes an outer membrane protein that confers complement resistance to serum-sensitive hosts.