Detecting envelope stress by monitoring ß-barrel assembly.

The cell envelope protects bacteria from their surroundings. Defects in its integrity or assembly are sensed by signal transduction systems, allowing cells to rapidly adjust. The Rcs phosphorelay responds to outer membrane (OM)- and peptidoglycan-related stress in enterobacteria. We elucidated how the OM lipoprotein RcsF, the upstream Rcs component, senses envelope stress and activates the signaling cascade. RcsF interacts with BamA, the major component of the ß-barrel assembly machinery. In growing cells, BamA continuously funnels RcsF through the ß-barrel OmpA, displaying RcsF on the cell surface. This process spatially separates RcsF from the downstream Rcs component, which we show is the inner membrane protein IgaA. The Rcs system is activated when BamA fails to bind RcsF and funnel it to OmpA. Newly synthesized RcsF then remains periplasmic, interacting with IgaA to activate the cascade. Thus RcsF senses envelope damage by monitoring the activity of the Bam machinery.

Disorder is a critical component of lipoprotein sorting in Gram-negative bacteria.

Gram-negative bacteria express structurally diverse lipoproteins in their cell envelope. Here, we find that approximately half of lipoproteins destined to the Escherichia coli outer membrane display an intrinsically disordered linker at their N terminus. Intrinsically disordered regions are common in proteins, but establishing their importance in vivo has remained challenging. As we sought to unravel how lipoproteins mature, we discovered that unstructured linkers are required for optimal trafficking by the Lol lipoprotein sorting system, whereby linker deletion re-routes three unrelated lipoproteins to the inner membrane. Focusing on the stress sensor RcsF, we found that replacing the linker with an artificial peptide restored normal outer-membrane targeting only when the peptide was of similar length and disordered. Overall, this study reveals the role played by intrinsic disorder in lipoprotein sorting, providing mechanistic insight into the biogenesis of these proteins and suggesting that evolution can select for intrinsic disorder that supports protein function.

A Comparison of Measured Airborne and Self-Reported Secondhand Smoke Exposure in the MADRES Pregnancy Cohort Study.

INTRODUCTION: Secondhand smoke (SHS) exposure during pregnancy is linked to adverse birth outcomes, such as low birth weight and preterm birth. While questionnaires are commonly used to assess SHS exposure, their ability to capture true exposure can vary, making it difficult for researchers to harmonize SHS measures. This study aimed to compare self-reported SHS exposure with measurements of airborne SHS in personal samples of pregnant women. METHODS: SHS was measured on 48-hour integrated personal PM2.5 Teflon filters collected from 204 pregnant women, and self-reported SHS exposure measures were obtained via questionnaires. Descriptive statistics were calculated for airborne SHS measures, and analysis of variance tests assessed group differences in airborne SHS concentrations by self-reported SHS exposure. RESULTS: Participants were 81% Hispanic, with a mean (SD) age of 28.2 (6.0) years. Geometric mean (SD) personal airborne SHS concentrations were 0.14 (9.41) µg/m3. Participants reporting lower education have significantly higher airborne SHS exposure (p=0.015). Mean airborne SHS concentrations were greater in those reporting longer duration with windows open in the home. There was no association between airborne SHS and self-reported SHS exposure; however, asking about the number of smokers nearby in the 48-hour monitoring period was most correlated with measured airborne SHS (Two+ smokers: 0.30µg/m3 vs. One: 0.12µg/m3 and Zero: 0.15µg/m3; p=0.230). CONCLUSIONS: Self-reported SHS exposure was not associated with measured airborne SHS in personal PM2.5 samples. This suggests exposure misclassification using SHS questionnaires and the need for harmonized and validated questions to characterize this exposure in health studies. IMPLICATIONS: This study adds to the growing body of evidence that measurement error is a major concern in pregnancy research, particularly in studies that rely on self-report questionnaires to measure secondhand smoke (SHS) exposure. The study introduces an alternative method of SHS exposure assessment using objective optical measurements, which can help improve the accuracy of exposure assessment. The findings emphasize the importance of using harmonized and validated SHS questionnaires in pregnancy health research to avoid biased effect estimates. This study can inform future research, practice, and policy development to reduce SHS exposure and its adverse health effects.

RoF-based indoor distributed antenna system that can simultaneously support 5G mmWave and 6G terahertz services.

The telecommunication society is paving the way toward ultra-high frequency regions, including the millimeter wave (mmWave) and sub-terahertz (sub-THz) bands. Such high-frequency electromagnetic waves induce a variety of physical constraints when they are used in wireless communications. Inevitably, the fiber-optic network is deeply embedded in the mobile network to resolve such challenges. In particular, the radio-over-fiber (RoF)-based distributed antenna system (DAS) can enhance the accessibility of next-generation mobile networks. The inherent benefits of RoF technology enhance the DAS network in terms of practicality and transmission performance by enabling it to support the 5G mmWave and 6G THz services simultaneously in a single optical transport link. Furthermore, the RoF allows the indoor network to be built based on the cascade architecture; thus, a service zone can be easily added on request. This study presents an RoF-based multi-service DAS network and experimentally investigates the feasibility of the proposed system.

Structural insight into the formation of lipoprotein-ß-barrel complexes.

The ß-barrel assembly machinery (BAM) inserts outer membrane ß-barrel proteins (OMPs) in the outer membrane of Gram-negative bacteria. In Enterobacteriacea, BAM also mediates export of the stress sensor lipoprotein RcsF to the cell surface by assembling RcsF-OMP complexes. Here, we report the crystal structure of the key BAM component BamA in complex with RcsF. BamA adopts an inward-open conformation, with the lateral gate to the membrane closed. RcsF is lodged deep within the lumen of the BamA barrel, binding regions proposed to undergo outward and lateral opening during OMP insertion. On the basis of our structural and biochemical data, we propose a push-and-pull model for RcsF export following conformational cycling of BamA, and provide a mechanistic explanation for how RcsF uses its interaction with BamA to detect envelope stress. Our data also suggest that the flux of incoming OMP substrates is involved in the control of BAM activity.

How the assembly and protection of the bacterial cell envelope depend on cysteine residues.

The cell envelope of Gram-negative bacteria is a multilayered structure essential for bacterial viability; the peptidoglycan cell wall provides shape and osmotic protection to the cell, and the outer membrane serves as a permeability barrier against noxious compounds in the external environment. Assembling the envelope properly and maintaining its integrity are matters of life and death for bacteria. Our understanding of the mechanisms of envelope assembly and maintenance has increased tremendously over the past two decades. Here, we review the major achievements made during this time, giving central stage to the amino acid cysteine, one of the least abundant amino acid residues in proteins, whose unique chemical and physical properties often critically support biological processes. First, we review how cysteines contribute to envelope homeostasis by forming stabilizing disulfides in crucial bacterial assembly factors (LptD, BamA, and FtsN) and stress sensors (RcsF and NlpE). Second, we highlight the emerging role of enzymes that use cysteine residues to catalyze reactions that are necessary for proper envelope assembly, and we also explain how these enzymes are protected from oxidative inactivation. Finally, we suggest future areas of investigation, including a discussion of how cysteine residues could contribute to envelope homeostasis by functioning as redox switches. By highlighting the redox pathways that are active in the envelope of Escherichia coli, we provide a timely overview of the assembly of a cellular compartment that is the hallmark of Gram-negative bacteria.

Distinct domains of Escherichia coli IgaA connect envelope stress sensing and down-regulation of the Rcs phosphorelay across subcellular compartments.

In enterobacteria, the Rcs system (Regulator of capsule synthesis) monitors envelope integrity and induces a stress response when damages occur in the outer membrane or in the peptidoglycan layer. Built around a two-component system, Rcs controls gene expression via a cascade of phosphoryl transfer reactions. Being particularly complex, Rcs also involves the outer membrane lipoprotein RcsF and the inner membrane essential protein IgaA (Intracellular growth attenuator). RcsF and IgaA, which are located upstream of the phosphorelay, are required for normal Rcs functioning. Here, we establish the stress-dependent formation of a complex between RcsF and the periplasmic domain of IgaA as the molecular signal triggering Rcs. Moreover, molecular dissection of IgaA reveals that its negative regulatory role on Rcs is mostly carried by its first N-terminal cytoplasmic domain. Altogether, our results support a model in which IgaA regulates Rcs activation by playing a direct role in the transfer of signals from the cell envelope to the cytoplasm. This remarkable feature further distinguishes Rcs from other envelope stress response systems.

Genome-Wide Analysis Reveals Mucociliary Remodeling of the Nasal Airway Epithelium Induced by Urban PM2.5.

Air pollution particulate matter <2.5 µm (PM2.5) exposure is associated with poor respiratory outcomes. Mechanisms underlying PM2.5-induced lung pathobiology are poorly understood but likely involve cellular and molecular changes to the airway epithelium. We extracted and chemically characterized the organic and water-soluble components of air pollution PM2.5 samples, then determined the whole transcriptome response of human nasal mucociliary airway epithelial cultures to a dose series of PM2.5 extracts. We found that PM2.5 organic extract (OE), but not water-soluble extract, elicited a potent, dose-dependent transcriptomic response from the mucociliary epithelium. Exposure to a moderate OE dose modified the expression of 424 genes, including activation of aryl hydrocarbon receptor signaling and an IL-1 inflammatory program. We generated an OE-response gene network defined by eight functional enrichment groups, which exhibited high connectivity through CYP1A1, IL1A, and IL1B. This OE exposure also robustly activated a mucus secretory expression program (>100 genes), which included transcriptional drivers of mucus metaplasia (SPDEF and FOXA3). Exposure to a higher OE dose modified the expression of 1,240 genes and further exacerbated expression responses observed at the moderate dose, including the mucus secretory program. Moreover, the higher OE dose significantly increased the MUC5AC/MUC5B gel-forming mucin expression ratio and strongly downregulated ciliated cell expression programs, including key ciliating cell transcription factors (e.g., FOXJ1 and MCIDAS). Chronic OE stimulation induced mucus metaplasia-like remodeling characterized by increases in MUC5AC+ secretory cells and MUC5AC mucus secretions. This epithelial remodeling may underlie poor respiratory outcomes associated with high PM2.5 exposure.

Demonstration of photonics-aided terahertz wireless transmission system with using silicon photonics circuit.

We experimentally demonstrate the use of silicon photonics circuit (SPC) in the simple and cost-effective photonics-aided Terahertz (THz) wireless transmission system. We perform theoretical investigation (with experimental confirmation) to understand that the system performance is more sensitive to the free space path loss (FSPL) at the THz wireless link than the SPC's insertion loss. The SPC, we design and fabricate, combines two incident optical carriers at different wavelengths and modulates one of two optical carriers with data to transfer, consequently reducing the system footprint that is indeed one of the key challenges that must be tackled for better practicability of the THz communication system. We perform experimental verification to show the feasibility of 40 Gb/s non-return-to-zero (NRZ) on-off-keying (OOK) signal transmission over 1.4 m wireless link for possibly its application in short-reach indoor wireless communication systems utilizing (sub-)THz frequency band such as, e.g., indoor WiFi, distributed antenna/radio systems, rack-to-rack data delivery, etc. The SPC could be further integrated with various photonic elements such as semiconductor optical amplifiers, laser diodes, and photo-mixers, which will enable the path towards all-photonic THz-wave synthesizers.

Demonstration of photonics-aided terahertz wireless transmission system with using silicon photonics circuit: erratum.

We present an erratum for our recent paper [Opt. Express 28, 23397 (2020)] to include funding information in the funding section.

Communication across the bacterial cell envelope depends on the size of the periplasm.

The cell envelope of gram-negative bacteria, a structure comprising an outer (OM) and an inner (IM) membrane, is essential for life. The OM and the IM are separated by the periplasm, a compartment that contains the peptidoglycan. The OM is tethered to the peptidoglycan via the lipoprotein, Lpp. However, the importance of the envelope's multilayered architecture remains unknown. Here, when we removed physical coupling between the OM and the peptidoglycan, cells lost the ability to sense defects in envelope integrity. Further experiments revealed that the critical parameter for the transmission of stress signals from the envelope to the cytoplasm, where cellular behaviour is controlled, is the IM-to-OM distance. Augmenting this distance by increasing the length of the lipoprotein Lpp destroyed signalling, whereas simultaneously increasing the length of the stress-sensing lipoprotein RcsF restored signalling. Our results demonstrate the physiological importance of the size of the periplasm. They also reveal that strict control over the IM-to-OM distance is required for effective envelope surveillance and protection, suggesting that cellular architecture and the structure of transenvelope protein complexes have been evolutionarily co-optimised for correct function. Similar strategies are likely at play in cellular compartments surrounded by 2 concentric membranes, such as chloroplasts and mitochondria.

Health impacts and cost-benefit analyses of surface O3 and PM2.5 over the U.S. under future climate and emission scenarios.

Health impacts of surface ozone (O3) and fine particulate matter (PM2.5) are of major concern worldwide. In this work, the Environmental Benefits Mapping and Analysis Program tool is applied to estimate the health and economic impacts of projected changes in O3 and PM2.5 in the U.S. in future (2046-2055) decade relative to current (2001-2010) decade under the Representative Concentration Pathway (RCP) 4.5 and 8.5 climate scenarios. Future annual-mean O3 reductions under RCP 4.5 prevent ~1,800 all-cause mortality, 761 respiratory hospital admissions (HA), and ~1.2 million school loss days annually, and result in economic benefits of ~16 billion, 29 million, and 132 million U.S. dollars (USD), respectively. By contrast, the projected future annual-mean O3 increases under RCP8.5 cause ~2,400 mortality, 941 respiratory HA, and ~1.6 million school loss days annually and result in economic disbenefits of ~21 billion, 36 million, and 175 million USD, respectively. Health benefits of reduced O3 double under RCP4.5 and health dis-benefits of increased O3 increase by 1.5 times under RCP8.5 in future with 2050 population and baseline incidence rate. Because of the reduction in projected future PM2.5 over CONUS under both scenarios, the annual avoided all-cause deaths, cardiovascular HA, respiratory HA, and work loss days are ~63,000 and ~83,000, ~5,300 and ~7,000, ~12,000 and ~15,000, and ~7.8 million and ~10 million, respectively, leading to economic benefits of ~560 and ~740 billion, ~240 and ~320 million, ~450 and ~590 million, and ~1,400 and ~1,900 million USD for RCP4.5 and 8.5, respectively. Health benefits of reduced PM2.5 for future almost double under both scenarios with the largest benefits in urban areas. RCP8.5 projects larger health and economic benefits due to a greater reduction in PM2.5 but with a warmer atmosphere and higher O3 pollution than RCP4.5. RCP4.5 leads to multiple-benefit goals including reduced O3 and PM2.5, reduced mortality and morbidity, and saved costs. Greater reduction in future PM2.5 under RCP4.5 should be considered to achieve larger multi-benefits.

4 × 4 MIMO architecture supporting IFoF-based analog indoor distributed antenna system for 5G mobile communications.

We demonstrate the analog indoor distributed antenna system (DAS) for 5G mobile communications that supports 4 × 4 multiple-input multiple-output (MIMO) configuration. For this, we exploit a pair of intermediate frequency over fiber (IFoF)-based analog optical links, transporting 32 frequency allocation (FA) 5G mobile signals (effectively ~4 GHz bandwidth). The analog optical link manifests its high fidelity in the measured characteristics: small gain variation (< ± 1 dB for the entire transmission bandwidth), low noise (<-136 dBm/Hz), and large dynamic range (spurious free dynamic range of >106 dB∙Hz2/3), subsequently providing superior error vector magnitude (EVM) performance (~2%) for a wide range of ambient temperatures (-20 ~60°C). Consequently, the IFoF-based 4 × 4 MIMO supporting analog indoor DAS is capable of providing record high peak throughput of 5.3 Gb/s for millimeter wave based 5G mobile communication system.

Investigation of transmission performance in multi-IFoF based mobile fronthaul with dispersion-induced intermixing noise mitigation.

We demonstrate the improvement of the transmission performance based on intermixing noise mitigation techniques in a multiple intermediate-frequency-over-fiber (IFoF) based mobile fronthaul. The interaction between fiber chromatic dispersion and frequency chirp of the directly modulated laser generates the second-order distortion that degrades the performance of multi-IFoF transmission system. To avoid second-order distortion, we use intermediate frequency (IF) spacing optimization and octave-confined frequency plan schemes in which intermixing noise would be generated in the out of signal band and would not affect the quality of transmitted signal. For bandwidth efficient transmission of radio signal over mobile fronthaul link, we employ the dispersion compensation technique to suppress the intermixing noise sufficiently. For realization of the multi-IFoF based mobile fronthaul, we experimentally investigate the transmission performances of 48-, 72- and 144-IF carriers of the long term evolution-advanced (LTE-A) signals mapped with 64-quadrature amplitude modulation (QAM). It is clearly observed that the intermixing noise is suppressed owing to dispersion compensation technique and overall system performances are improved by IF spacing optimization and octave-confined frequency plan. As a result, we successfully transmit 144-IF carriers of the LTE-A signal with less than 8% error vector magnitude (EVM) over 20-km single-mode fiber (SMF) within only 3 GHz bandwidth.

Medialized repair for retracted rotator cuff tears.

BACKGROUND: The purpose of this study was to evaluate the functional outcomes of medialized rotator cuff repair and the continuity of repaired tendon in chronic retracted rotator cuff tears. METHODS: Thirty-five consecutive patients were selected from 153 cases that underwent arthroscopic rotator cuff repair for more than medium-sized posterosuperior rotator cuff tears between July 2009 and July 2012 performed with the medialized repair. All cases were available for at least 2 years of postoperative follow-up. The visual analog scale of pain, muscle strength, Constant score, American Shoulder and Elbow Surgeons (ASES) score, and University of California-Los Angeles score were evaluated. RESULTS: At the final follow-up, all clinical outcomes were significantly improved. The visual analog scale score for pain improved from 6 ± 1 preoperatively to 2 ± 1 postoperatively. The range of motion increased from preoperatively to postoperatively: active forward elevation, from 134° ± 49° to 150° ± 16°; active external rotation at the side, from 47° ± 15° to 55° ± 10°; and active internal rotation, from L3 to L1. The shoulder score also improved: Constant score, from 53.5 ± 16.7 to 79 ± 10; American Shoulder and Elbow Surgeons score, from 51 ± 15 to 82 ± 8; and University of California-Los Angeles score, from 14 ± 4 to 28 ± 4. The retear cases at the final follow-up were 6 (17%). CONCLUSIONS: Medialized repair may be useful in cases in which anatomic bone-to-tendon repair would be difficult because of the excessive tension of the repaired tendon and a torn tendon that does not reach the anatomic insertion.

Improvement of the transmission performance in multi-IF-over-fiber mobile fronthaul by using tone-reservation technique.

We demonstrate the improvement of the transmission performance based on tone-reservation technique in a multiple intermediate-frequency-over-fiber (IFoF) based mobile fronthaul. The tone-reservation technique can suppress nonlinear distortion by eliminating the high peak components of orthogonal frequency-division multiplexing (OFDM) signal. To prevent the regrowth of peak, we employ tone-reservation after multiplexing IF carriers. Furthermore, we use an out-of-band signal as the reserved tones to avoid any modification of a mobile signal. The impact of the number of IF carriers on peak-to-average power ratio (PAPR) characteristics is presented via numerical simulation. For the multi-IFoF based mobile fronthaul, we experimentally investigate the transmission performance of 36-IF carriers of the long term evolution-advanced (LTE-A) signals mapped with 64-quadrature amplitude modulation (QAM). It is clearly observed that the clipping-induced nonlinear distortion is dramatically suppressed by using tone-reservation. As a result, the transmission performance of 36-IF carriers of the LTE-A signal is improved by an error-vector-magnitude (EVM) of 4% (from 9.7% to 5.7%) after 20-km transmission.

Development of a Small-Sized, Flexible, and Insertable Fiber-Optic Radiation Sensor for Gamma-Ray Spectroscopy.

We fabricated a small-sized, flexible, and insertable fiber-optic radiation sensor (FORS) that is composed of a sensing probe, a plastic optical fiber (POF), a photomultiplier tube (PMT)-amplifier system, and a multichannel analyzer (MCA) to obtain the energy spectra of radioactive isotopes. As an inorganic scintillator for gamma-ray spectroscopy, a cerium-doped lutetium yttrium orthosilicate (LYSO:Ce) crystal was used and two solid-disc type radioactive isotopes with the same dimensions, cesium-137 (Cs-137) and cobalt-60 (Co-60), were used as gamma-ray emitters. We first determined the length of the LYSO:Ce crystal considering the absorption of charged particle energy and measured the gamma-ray energy spectra using the FORS. The experimental results demonstrated that the proposed FORS can be used to discriminate species of radioactive isotopes by measuring their inherent energy spectra, even when gamma-ray emitters are mixed. The relationship between the measured photon counts of the FORS and the radioactivity of Cs-137 was subsequently obtained. The amount of scintillating light generated from the FORS increased by increasing the radioactivity of Cs-137. Finally, the performance of the fabricated FORS according to the length and diameter of the POF was also evaluated. Based on the results of this study, it is anticipated that a novel FORS can be developed to accurately measure the gamma-ray energy spectrum in inaccessible locations such as narrow areas and holes.

Enhanced cytotoxic and genotoxic effects of gadolinium following ELF-EMF irradiation in human lymphocytes.

There are many studies of Gd nephrotoxicity and neurotoxicity, whereas research on cyto- and genotoxicity in normal human lymphocytes is scarce. It is important to investigate the effect of extremely low-frequency electromagnetic fields (ELF-EMF) on Gd toxicity, as patients are co-exposed to Gd and ELF-EMF generated by MRI scanners. We investigated the cytotoxicity and genotoixcity of Gd and the possible enhancing effect of ELF-EMF on Gd toxicity in cultured human lymphocytes by performing a micronuclei (MN) assay, trypan blue dye exclusion, single cell gel electrophoresis, and apoptosis analyses using flow cytometry. Isolated lymphocytes were exposed to 0.2-1.2 mM of Gd only or in combination with a 60-Hz ELF-EMF of 0.8-mT field strength. Exposing human lymphocytes to Gd resulted in a concentration- and time-dependent decrease in cell viability and an increase in MN frequency, single strand DNA breakage, apoptotic cell death, and ROS production. ELF-EMF (0.8 mT) exposure also increased cell death, MN frequency, olive tail moment, and apoptosis induced by Gd treatment alone. These results suggest that Gd induces DNA damage and apoptotic cell death in human lymphocytes and that ELF-EMF enhances the cytotoxicity and genotoxicity of Gd.

A fiber-optic sensor using an aqueous solution of sodium chloride to measure temperature and water level simultaneously.

A fiber-optic sensor system using a multiplexed array of sensing probes based on an aqueous solution of sodium chloride (NaCl solution) and an optical time-domain reflectometer (OTDR) for simultaneous measurement of temperature and water level is proposed. By changing the temperature, the refractive index of the NaCl solution is varied and Fresnel reflection arising at the interface between the distal end of optical fiber and the NaCl solution is then also changed. We measured the modified optical power of the light reflected from the sensing probe using a portable OTDR device and also obtained the relationship between the temperature of water and the optical power. In this study, the water level was simply determined by measuring the signal difference of the optical power due to the temperature difference of individual sensing probes placed inside and outside of the water. In conclusion, we demonstrate that the temperature and water level can be obtained simultaneously by measuring optical powers of light reflected from sensing probes based on the NaCl solution. It is anticipated that the proposed fiber-optic sensor system makes it possible to remotely monitor the real-time change of temperature and water level of the spent fuel pool during a loss of power accident.

Measurement of entrance surface dose on an anthropomorphic thorax phantom using a miniature fiber-optic dosimeter.

A miniature fiber-optic dosimeter (FOD) system was fabricated using a plastic scintillating fiber, a plastic optical fiber, and a multi-pixel photon counter to measure real-time entrance surface dose (ESD) during radiation diagnosis. Under varying exposure parameters of a digital radiography (DR) system, we measured the scintillating light related to the ESD using the sensing probe of the FOD, which was placed at the center of the beam field on an anthropomorphic thorax phantom. Also, we obtained DR images using a flat panel detector of the DR system to evaluate the effects of the dosimeter on image artifacts during posteroanterior (PA) chest radiography. From the experimental results, the scintillation output signals of the FOD were similar to the ESDs including backscatter simultaneously obtained using a semiconductor dosimeter. We demonstrated that the proposed miniature FOD can be used to measure real-time ESDs with minimization of DR image artifacts in the X-ray energy range of diagnostic radiology.