Genetic determinants of ammonia-induced acute lung injury in mice.

In this study, a genetically diverse panel of 43 mouse strains was exposed to ammonia, and genome-wide association mapping was performed employing a single-nucleotide polymorphism (SNP) assembly. Transcriptomic analysis was used to help resolve the genetic determinants of ammonia-induced acute lung injury. The encoded proteins were prioritized based on molecular function, nonsynonymous SNP within a functional domain or SNP within the promoter region that altered expression. This integrative functional approach revealed 14 candidate genes that included Aatf, Avil, Cep162, Hrh4, Lama3, Plcb4, and Ube2cbp, which had significant SNP associations, and Aff1, Bcar3, Cntn4, Kcnq5, Prdm10, Ptcd3, and Snx19, which had suggestive SNP associations. Of these genes, Bcar3, Cep162, Hrh4, Kcnq5, and Lama3 are particularly noteworthy and had pathophysiological roles that could be associated with acute lung injury in several ways.

Toll interacting protein protects bronchial epithelial cells from bleomycin-induced apoptosis.

Idiopathic pulmonary fibrosis (IPF) is characterized by altered epithelial cell phenotypes, which are associated with myofibroblast accumulation in the lung. Atypical alveolar epithelial cells in IPF express molecular markers of airway epithelium. Polymorphisms within and around Toll interacting protein (TOLLIP) are associated with the susceptibility to IPF and mortality. However, the functional role of TOLLIP in IPF is unknown. Using lung tissues from IPF and control subjects, we showed that expression of TOLLIP gene in the lung parenchyma is globally lower in IPF compared to controls. Lung cells expressing significant levels of TOLLIP include macrophages, alveolar type II, and basal cells. TOLLIP protein expression is lower in the parenchyma of IPF lungs but is expressed in the atypical epithelial cells of the distal fibrotic regions. Using overexpression and silencing approaches, we demonstrate that TOLLIP protects cells from bleomycin-induced apoptosis using primary bronchial epithelial cells and BEAS-2B cells. The protective effects are mediated by reducing mitochondrial reactive oxygen species (ROS) levels and upregulating autophagy. Therefore, global downregulation of the TOLLIP gene in IPF lungs may predispose injured lung epithelial cells to apoptosis and to the development of IPF.

Pulmonary Inflammation and KRAS Mutation in Lung Cancer.

Chronic lung infection and lung cancer are two of the most important pulmonary diseases. Respiratory infection and its associated inflammation have been increasingly investigated for their role in increasing the risk of respiratory diseases including chronic obstructive pulmonary disease (COPD) and lung cancer. Kirsten rat sarcoma viral oncogene (KRAS) is one of the most important regulators of cell proliferation, differentiation, and survival. KRAS mutations are among the most common drivers of cancer. Lung cancer harboring KRAS mutations accounted for ~25% of the incidence but the relationship between KRAS mutation and inflammation remains unclear. In this chapter, we will describe the roles of KRAS mutation in lung cancer and how elevated inflammatory responses may increase KRAS mutation rate and create a vicious cycle of chronic inflammation and KRAS mutation that likely results in persistent potentiation for KRAS-associated lung tumorigenesis. We will discuss in this chapter regarding the studies of KRAS gene mutations in specimens from lung cancer patients and in animal models for investigating the role of inflammation in increasing the risk of lung tumorigenesis driven primarily by oncogenic KRAS.

A Rare Mutation in SPLUNC1 Affects Bacterial Adherence and Invasion in Meningococcal Disease.

BACKGROUND: Neisseria meningitidis (Nm) is a nasopharyngeal commensal carried by healthy individuals. However, invasive infections occurs in a minority of individuals, with devastating consequences. There is evidence that common polymorphisms are associated with invasive meningococcal disease (IMD), but the contributions of rare variants other than those in the complement system have not been determined. METHODS: We identified familial cases of IMD in the UK meningococcal disease study and the European Union Life-Threatening Infectious Disease Study. Candidate genetic variants were identified by whole-exome sequencing of 2 patients with familial IMD. Candidate variants were further validated by in vitro assays. RESULTS: Exomes of 2 siblings with IMD identified a novel heterozygous missense mutation in BPIFA1/SPLUNC1. Sequencing of 186 other nonfamilial cases identified another unrelated IMD patient with the same mutation. SPLUNC1 is an innate immune defense protein expressed in the nasopharyngeal epithelia; however, its role in invasive infections is unknown. In vitro assays demonstrated that recombinant SPLUNC1 protein inhibits biofilm formation by Nm, and impedes Nm adhesion and invasion of human airway cells. The dominant negative mutant recombinant SPLUNC1 (p.G22E) showed reduced antibiofilm activity, increased meningococcal adhesion, and increased invasion of cells, compared with wild-type SPLUNC1. CONCLUSIONS: A mutation in SPLUNC1 affecting mucosal attachment, biofilm formation, and invasion of mucosal epithelial cells is a new genetic cause of meningococcal disease.

Synergistic Biophysical Techniques Reveal Structural Mechanisms of Engineered Cationic Antimicrobial Peptides in Lipid Model Membranes.

In the quest for new antibiotics, two novel engineered cationic antimicrobial peptides (eCAPs) have been rationally designed. WLBU2 and D8 (all 8 valines are the d-enantiomer) efficiently kill both Gram-negative and -positive bacteria, but WLBU2 is toxic and D8 nontoxic to eukaryotic cells. We explore protein secondary structure, location of peptides in six lipid model membranes, changes in membrane structure and pore evidence. We suggest that protein secondary structure is not a critical determinant of bactericidal activity, but that membrane thinning and dual location of WLBU2 and D8 in the membrane headgroup and hydrocarbon region may be important. While neither peptide thins the Gram-negative lipopolysaccharide outer membrane model, both locate deep into its hydrocarbon region where they are primed for self-promoted uptake into the periplasm. The partially α-helical secondary structure of WLBU2 in a red blood cell (RBC) membrane model containing 50 % cholesterol, could play a role in destabilizing this RBC membrane model causing pore formation that is not observed with the D8 random coil, which correlates with RBC hemolysis caused by WLBU2 but not by D8.

15-Lipoxygenase 1 in nasal polyps promotes CCL26/eotaxin 3 expression through extracellular signal-regulated kinase activation.

BACKGROUND: 15-Lipoxygenase 1 (15LO1) is expressed in airway epithelial cells in patients with type 2-high asthma in association with eosinophilia. Chronic rhinosinusitis with nasal polyps (CRSwNP) is also associated with type 2 inflammation and eosinophilia. CCL26/eotaxin 3 has been reported to be regulated by 15LO1 in lower airway epithelial cells. However, its relation to 15LO1 in patients with CRSwNP or mechanisms for its activation are unclear. OBJECTIVE: We sought to evaluate 15LO1 and CCL26 expression in nasal epithelial cells (NECs) from patients with CRSwNP and healthy control subjects (HCs) and determine whether 15LO1 regulates CCL26 in NECs through extracellular signal-regulated kinase (ERK) activation. METHODS: 15LO1, CCL26, and phosphorylated ERK were evaluated in NECs from patients with CRSwNP and HCs. 15LO1/CCL26 and CCL26/cytokeratin 5 were colocalized by means of immunofluorescence. IL-13-stimulated NECs were cultured at an air-liquid interface with or without 15-lipoxygenase 1 gene (ALOX15) Dicer-substrate short interfering RNAs (DsiRNA) transfection, a specific 15LO1 enzymatic inhibitor, and 2 ERK inhibitors. Expression of 15LO1 and CCL26 mRNA and protein was analyzed by using quantitative RT-PCR, Western blotting, and ELISA. RESULTS: 15LO1 expression was increased in nasal polyp (NP) epithelial cells compared with middle turbinate epithelial cells from patients with CRSwNP and HCs. 15LO1 expression correlated with CCL26 expression and colocalized with CCL26 expression in basal cells of the middle turbinate and NPs from patients with CRSwNP. In primary NECs in vitro, IL-13 induced 15LO1 and CCL26 expression. 15LO1 knockdown and inhibition decreased IL-13-induced ERK phosphorylation and CCL26 expression. ERK inhibition (alone) similarly decreased IL-13-induced CCL26. Phosphorylated ERK expression was increased in NECs from CRSwNP subjects and positively correlated with both 15LO1 and CCL26 expression. CONCLUSIONS: 15LO1 expression is increased in NP epithelial cells and contributes to CCL26 expression through ERK activation. 15LO1 could be considered a novel therapeutic target for CRSwNP.

[Retinal toxicity study of intravitreal ganciclovir in albino rabbit].

Objective: To elevated the retinal toxicity of intravitreal ganciclovir in albino rabbit eyes. Methods: Experimental study. Twenty-four New Zealand albino rabbits (forty-eight eyes) were divided into four groups by random. Three groups were prepared for ganciclovir experiment, named A, B, C. Each group received intravitreal injection ganciclovir dose at 400 µg/0.05 ml, 2 mg/0.05 ml and 5 mg/0.05 ml respectively. The other group named D served as a control accepted intravitreal injection 0.9% normal saline 0.1 ml. Before and after 1, 2 and 4 weeks, flicker full field electroretina gram (ERG) was recorded. After 1, 2 and 4 weeks light and electron microscopic tests were recorded for further toxicity study. Results: There was significant difference in amplitude of maximal combined response a wave in one week(χ(2)=8.319, P=0.04), and pairwise comparison the 5 mg group (140.50 µV) was significantly lower than the control group (165.00 µV) (χ(2)=-2.830, P=0.028). Maximal combined response b wave in four weeks(χ(2)=-10.626, P=0.014), and pairwise comparison the 5 mg group (261.50 µV) was significantly lower than the control group (398.00 µV) (χ(2)=-2.973, P=0.018). 30 Hz flicker response in one, two and four weeks(χ(2)=17.589, 8.225, 8.997, P=0.001, 0.042, 0.02), and pairwise comparison the 5 mg group (71.3µV, 106.00µV, 63.60µV) was significantly lower than the control group (118.50µV, 129.00µV, 116.50µV) (χ(2)=-4.142, -2.826, -2.713, P=0.000, 0.028, 0.040). There was no histologic retinal toxicity evidence of group 400 µg and control group observed by light microscopy in any stage of the study. Histologic changes of group 2 mg four week later, group 5 mg two and four week later include inner nuclear layer loose arranged, nuclear of ganglia were widened and outer plexiform layer stained less in four week later. By electron microscopic observation, the ultrastructure of retina changed to different degrees and became worse in each experimental group with significant mitochondrial swelling and hydropic changes were seen in the inner segments of photoreceptors, loosely arranged and disordered in the outer segment of photoreceptors four weeks later. Conclusions: The retinal function and morphology were normal in group 400 µg. Group 2 mg and 5 mg had retinal toxicity, and 5 mg was more severe. Therefore, the clinical application of ganciclovir in the treatment of acute retinal necrosis (ARN) should select the minimum effective dose to avoid the occurrence of retinal toxicity. (Chin J Ophthalmol, 2020, 56:279-285).

Elastic behavior of model membranes with antimicrobial peptides depends on lipid specificity and d-enantiomers.

In an effort to provide new treatments for the global crisis of bacterial resistance to current antibiotics, we have used a rational approach to design several new antimicrobial peptides (AMPs). The present study focuses on 24-mer WLBU2 and its derivative, D8, with the amino acid sequence, RRWVRRVRRWVRRVVRVVRRWVRR. In D8, all of the valines are the d-enantiomer. We use X-ray low- and wide-angle diffuse scattering data to measure elasticity and lipid chain order. We show a good correlation between in vitro bacterial killing efficiency and both bending and chain order behavior in bacterial lipid membrane mimics; our results suggest that AMP-triggered domain formation could be the mechanism of bacterial killing in both Gram-positive and Gram-negative bacteria. In red blood cell lipid mimics, D8 stiffens and orders the membrane, while WLBU2 softens and disorders it, which correlate with D8's harmless vs. WLBU2's toxic behavior in hemolysis tests. These results suggest that elasticity and chain order behavior can be used to predict mechanisms of bactericidal action and toxicity of new AMPs.

[The ocular manifestations of human immunodeficiency virus and syphilis coinfection].

Objective: To investigate the ocular manifestations of human immunodeficiency virus (HIV) and syphilis coinfection. Methods: A retrospective analysis of the ocular manifestations was carried out in 27 patients (54 eyes) diagnosed as syphilis and HIV coinfection by the Department of Infectious Medicine in Peking Union Medical College Hospital during the years of 2006-2017. The research included 26 males and 1 female, aging from 24 to 76 years old, with a mean age of 40.40±12.94 years old. Ocular anterior segments were examined with slit-lamp microscope. Fundus examinations were conducted with papillary dilation, fundus photography. Results: At the first visit, there were 2 eyes without light perception, 4 eyes with light perception, 1 eye with hand movement, 1 eye with finger counting, 2 eyes with 0.01-0.09 eyesight, 8 eyes with 0.1-0.2 eyesight, 12 eyes with 0.25-0.4 eyesight, 15 eyes with 0.5-0.9 eyesight, 9 eyes with 1.0-1.5 eyesight. Among the 27 patients (54 eyes) coinfected with HIV and syphilis, keratic precipitates were identified in 20 eyes, aqueous flare positive in 20 eyes, float positive in 15 eyes, and iris posterior synechias in 7 eyes. Nineteen eyes were diagnosed as syphilis uveitis, including 2 eyes with syphilis anterior uveitis and 17 eyes with syphilis panuveitis, among which, vitreous inflammatory opacity was observed in all 19 eyes, disk atrophy in 2 eyes, optic edema in 1 eye, vitreous hemorrhage in 1 eye, retinal detachment in 2 eyes, retinal hemorrhage and white vein in 1 eye. In addition, 8 eyes were diagnosed as HIV retinopathy, all manifested as cotton-wool spot. Among the 8 eyes, 4 were diagnosed as cytomegalovirus retinitis, 3 showed retinal yellow-white lesions, and 1 was in late phase which showed retinal pigmentation. The incidence of both HIV and syphilis coinfection patients and male homosexuality population increased. The most common ocular manifestation of HIV and syphilis coinfection was syphilis panuveitis. Six patients first visited the Department of Ophthalmology, and were then diagnosed as HIV and syphilis coinfection. Conclusions: The ocular manifestations of HIV and syphilis coinfection are diversified, which can be manifested as fundus necrotic lesions as well as anterior and posterior inflammatory. For HIV positive patients, syphilis serologic test should be routinely performed. The same, syphilis positive patients should be tested for HIV serum antibodies, in order to improve the diagnosis level of HIV/syphilis coinfection and give timely etiological treatment, which is of vital importance for saving visual acuity. (Chin J Ophthalmol, 2019, 55:267-272).

Association of intraocular pressure-related factors and retinal vessel diameter with optic disc rim area in subjects with and without primary open angle glaucoma.

IMPORTANCE: The data may support the notion that the intra-ocular pressure (IOP)-related factors and vascular factors were implicated concurrently in glaucomatous optic nerve damage. BACKGROUND: To study the association of intraocular pressure (IOP)-related factors, IOP, trans-lamina cribrosa pressure difference (TLCPD), cerebrospinal fluid pressure (CSFP) and retinal vessel diameters (RVD), central retinal arteriolar equivalent (CRAE), central retinal venular equivalent (CRVE) with neuro-retinal rim area (RA). DESIGN: A population-based, cross-sectional study. PARTICIPANTS: A total of 6830 people aged 30 years and over. METHODS: All participants underwent a comprehensive eye examination, fundus photograph-based measurements of RVD and Heidelberg retinal tomogram (HRT) measurement of optic disc. MAIN OUTCOME MEASURES: RA, CRAE, CRVE, IOP, body mass index (BMI), CSFP and TLCPD. RESULTS: Primary open-angle glaucoma (POAG) was diagnosed using two separate methods: 67 from expert consensus, and 125 from the International Society of Geographical and Epidemiologic Ophthalmology (ISGEO) classification. After excluding of those with high myopia or without gradable HRT images, 4194 non-glaucoma and 40 POAG were analysed for determinants of RA. On multivariable analysis determinants of reduced RA were POAG (P < 0.001), higher IOP (P = 0.03), higher refractive error (P < 0.01), longer axial length (P = 0.01), CRVE (P < 0.001), lower BMI (P = 0.015), older age (P < 0.001) smaller disc area (P < 0.001) and higher TLCPD (P = 0.03). When age and/or BMI were omitted from the model, reduced RA was also associated with lower CSFP (P < 0.001). CONCLUSIONS AND RELEVANCE: Reduced RA is associated with narrow CRVE and higher IOP or lower CSFP. The data supports the concurrent role of IOP-related and vascular factors in glaucomatous optic nerve damage.

[Research progress on the diagnosis and treatment of acute retinal necrosis].

Acute retinal necrosis is an uncommon but devastating and potentially blinding ophthalmopathy featured with unfavorable prognosis as the lesions tend to develop rapidly in a circumferential way in few days. Therefore timely diagnosis and early treatment are of great significance. The polymerase chain reaction test on intraocular fluid can improve specificity and sensitivity of the etiological detection for acute retinal necrosis. Etiology results included in the diagnostic criteria is favorable for etiological diagnosis. Current treatment for acute retinal necrosis is generally based on systemic antiviral therapy, and supported by hormone and antithrombotic drugs, while surgical treatment should be considered when the disease progresses. This article reviews the research progress on the diagnosis and treatment of acute retinal necrosis in an effort to improve the diagnosis and treatment performance of acute retinal necrosis. (Chin J Ophthalmol, 2018, 54: 306-311).

[The relationship between optical coherence tomography performance and visual acuity of acute retinal necrosis].

Objective: To analyze the relationship between optical coherence tomography(OCT) performance and visual acuity of patients with acute retinal necrosis (ARN). Methods: Retrospective analysis was performed on the patients diagnosed with ARN at the ophthalmology department of Peking union hospital during October 2011 and May 2016. Fourteen patients (15 eyes), 9 males and 5 females, whose anterior and posterior inflammation disappeared and the retinal necrosis lesion in the fundus of the eye subsided were included. The mean age was (41.6±12.2) years. All patients underwent careful examinations including best corrected visual acuity (BCVA), slit-lamp microscope, indirect ophthalmoscope, color fundus picture, fundus fluorescein angiography (FFA) and OCT (results of their last consultations). Results: BCVA: 8 eyes were increased, 2 eyes were unchanged and 5 eyes were decreased at the last visit; light perception (LP) 1 eye, finger count (FC) 1 eye, 3 eyes of 0.01 to 0.1, 6 eyes of 0.15 to 0.25. The inflammatory reaction in the anterior segment of 14 eyes disappeared, while the inflammatory reaction of the remaining 1 eye was relieved. The fundus lesions of all 15 eyes disappeared. According to OCT results: five eyes (5/15) exhibited normal macula area, among the 5 eyes, 4 eyes are of 0.15 to 0.25 visual acuity, and 1 eye is of 0.5 visual acuity;macular epiretinal membrane is present in three eyes (3/15), of which the visual acuity is 0.02, 0.25 and 0.3 respectively macula edema is present in three eyes (3/15), among the 3 eyes, 1 eye (visual acuity of 0.01)showed thickening of neurosensory retina, cystoid change of fovea and several fluid dark areas, the other 2 eyes (visual acuity of 0.02 and 0.5 respectively) showed small diffuse fluid dark area in the neurosensory retina;atrophy of neurosensory retina and absence of IS/OS was found in four eyes (4/15), among the 4 eyes, the visual acuity of 3 is below 0.01, and the other 1 eye is of 0.08 visual acuity. Conclusions: The OCT performance of stationary phase of ARN tends to be positively correlated with the visual acuity of patients. The prognosis of visual acuity of the patients whose OCT results showed atrophy of neurosensory retina and absence of IS/OS is poor. (Chin J Ophthalmol, 2018, 54: 369-374).

[Application of optical coherence tomography angiography in ophthalmology].

Optical coherence tomography angiography (OCTA) is a new technology of angiography in recent years. It has advantages of the optical coherence tomography (OCT), moreover, it can hierarchically observe the vascular morphology of the retina and choroid. By using the pseudo-color, abnormal vascular structure can be distinguished from normal vascular structure of the retina. With the fusion of the OCTA image and the regular OCT image, the structure and blood flow information can be obtained at the same time. This technology provides another method of examination for clinical ophthalmology, especially for the diagnosis of retinal diseases. This review will analyze and summarize the operating principle of OCTA, its application in ophthalmology, as well as its advantages and limitations. (Chin J Ophthalmol, 2017, 53: 65-72).

[Current research of sympathetic ophthalmia].

Sympathetic ophthalmia (SO) is a rare, bilateral, non-necrotizing, granulomatous uveitis that usually occurs after open ocular injury or intraocular surgery. The pathophysiology is not clearly understood, but generally SO is an immediate hypersensitivity mediated by T lymphocytes which are related to ocular tissue antigens. The main histopathological features are granulation tissues composed of lymphocytes, macrophages and multinucleated giant cells. The clinical manifestations are different from person to person, which might be mild or severe. Although it could be presented with anterior uveitis, intermediate uveitis and posterior uveitis, panuveitis is the most common sign. The ophthalmic examinations, such as fundus fluorescein angiography, optical coherence tomography and B-scan, could be used to observe the patients' conditions and monitor the therapeutic effect. The main treatment of SO is medical therapy with corticosteroids, immunomodulators and biomodulators. Topical drug administration, including intravitreal injection of triamcinolone acetonide and implantation of a fluocinolone acetonide implant, can be considered. There is controversy about whether enucleation or evisceration is more appropriate and when the procedure should be done. The prognosis of SO could be poor. SO is liable to deteriorate and may lead to blindness. This article reviews the etiology, mechanisms, histopathology, clinical characteristics, diagnosis and treatment of SO. (Chin J Ophthalmol, 2017, 53:778-782).

Structural Features Essential to the Antimicrobial Functions of Human SPLUNC1.

SPLUNC1 is an abundantly secreted innate immune protein in the mammalian respiratory tract that exerts bacteriostatic and antibiofilm effects, binds to lipopolysaccharide (LPS), and acts as a fluid-spreading surfactant. Here, we unravel the structural elements essential for the surfactant and antimicrobial functions of human SPLUNC1 (short palate lung nasal epithelial clone 1). A unique α-helix (α4) that extends from the body of SPLUNC1 is required for the bacteriostatic, surfactant, and LPS binding activities of this protein. Indeed, we find that mutation of just four leucine residues within this helical motif to alanine is sufficient to significantly inhibit the fluid spreading abilities of SPLUNC1, as well as its bacteriostatic actions against Gram-negative pathogens Burkholderia cenocepacia and Pseudomonas aeruginosa. Conformational flexibility in the body of SPLUNC1 is also involved in the bacteriostatic, surfactant, and LPS binding functions of the protein as revealed by disulfide mutants introduced into SPLUNC1. In addition, SPLUNC1 exerts antibiofilm effects against Gram-negative bacteria, although α4 is not involved in this activity. Interestingly, though, the introduction of surface electrostatic mutations away from α4 based on the unique dolphin SPLUNC1 sequence, and confirmed by crystal structure, is shown to impart antibiofilm activity against Staphylococcus aureus, the first SPLUNC1-dependent effect against a Gram-positive bacterium reported to date. Together, these data pinpoint SPLUNC1 structural motifs required for the antimicrobial and surfactant actions of this protective human protein.

Esculentin-1a-Derived Peptides Promote Clearance of Pseudomonas aeruginosa Internalized in Bronchial Cells of Cystic Fibrosis Patients and Lung Cell Migration: Biochemical Properties and a Plausible Mode of Action.

Pseudomonas aeruginosa is the major microorganism colonizing the respiratory epithelium in cystic fibrosis (CF) sufferers. The widespread use of available antibiotics has drastically reduced their efficacy, and antimicrobial peptides (AMPs) are a promising alternative. Among them, the frog skin-derived AMPs, i.e., Esc(1-21) and its diastereomer, Esc(1-21)-1c, have recently shown potent activity against free-living and sessile forms of P. aeruginosa Importantly, this pathogen also escapes antibiotics treatment by invading airway epithelial cells. Here, we demonstrate that both AMPs kill Pseudomonas once internalized into bronchial cells which express either the functional or the ΔF508 mutant of the CF transmembrane conductance regulator. A higher efficacy is displayed by Esc(1-21)-1c (90% killing at 15 µM in 1 h). We also show the peptides' ability to stimulate migration of these cells and restore the induction of cell migration that is inhibited by Pseudomonas lipopolysaccharide when used at concentrations mimicking lung infection. This property of AMPs was not investigated before. Our findings suggest new therapeutics that not only eliminate bacteria but also can promote reepithelialization of the injured infected tissue. Confocal microscopy indicated that both peptides are intracellularly localized with a different distribution. Biochemical analyses highlighted that Esc(1-21)-1c is significantly more resistant than the all-l peptide to bacterial and human elastase, which is abundant in CF lungs. Besides proposing a plausible mechanism underlying the properties of the two AMPs, we discuss the data with regard to differences between them and suggest Esc(1-21)-1c as a candidate for the development of a new multifunctional drug against Pseudomonas respiratory infections.

Differential Susceptibilities of Human Lung Primary Cells to H1N1 Influenza Viruses.

UNLABELLED: Human alveolar epithelial cells (AECs) and alveolar macrophages (AMs) are the first lines of lung defense. Here, we report that AECs are the direct targets for H1N1 viruses that have circulated since the 2009 pandemic (H1N1pdm09). AMs are less susceptible to H1N1pdm09 virus, but they produce significantly more inflammatory cytokines than AECs from the same donor. AECs form an intact epithelial barrier that is destroyed by H1N1pdm09 infection. However, there is significant variation in the cellular permissiveness to H1N1pdm09 infection among different donors. AECs from obese donors appear to be more susceptible to H1N1pdm09 infection, whereas gender, smoking history, and age do not appear to affect AEC susceptibility. There is also a difference in response to different strains of H1N1pdm09 viruses. Compared to A/California04/09 (CA04), A/New York/1682/09 (NY1682) is more infectious and causes more epithelial barrier injury, although it stimulates less cytokine production. We further determined that a single amino acid residue substitution in NY1682 hemagglutinin is responsible for the difference in infectivity. In conclusion, this is the first study of host susceptibility of human lung primary cells and the integrity of the alveolar epithelial barrier to influenza. Further elucidation of the mechanism of increased susceptibility of AECs from obese subjects may facilitate the development of novel protection strategies against influenza virus infection. IMPORTANCE: Disease susceptibility of influenza is determined by host and viral factors. Human alveolar epithelial cells (AECs) form the key line of lung defenses against pathogens. Using primary AECs from different donors, we provided cellular level evidence that obesity might be a risk factor for increased susceptibility to influenza. We also compared the infections of two closely related 2009 pandemic H1N1 strains in AECs from the same donor and identified a key viral factor that affected host susceptibility, the dominance of which may be correlated with disease epidemiology. In addition, primary human AECs can serve as a convenient and powerful model to investigate the mechanism of influenza-induced lung injury and determine the effect of genetic and epigenetic factors on host susceptibility to pandemic influenza virus infection.

Influenza-induced type I interferon enhances susceptibility to gram-negative and gram-positive bacterial pneumonia in mice.

Suppression of type 17 immunity by type I interferon (IFN) during influenza A infection has been shown to enhance susceptibility to secondary bacterial pneumonia. Although this mechanism has been described in coinfection with gram-positive bacteria, it is unclear whether similar mechanisms may impair lung defense against gram-negative infections. Furthermore, precise delineation of the duration of type I IFN-associated susceptibility to bacterial infection remains underexplored. Therefore, we investigated the effects of preceding influenza A virus infection on subsequent challenge with the gram-negative bacteria Escherichia coli or Pseudomonas aeruginosa and the temporal association between IFN expression with susceptibility to Staphylococcus aureus challenge in a mouse model of influenza and bacterial coinfection. Here we demonstrate that preceding influenza A virus led to increased lung E. coli and P. aeruginosa bacterial burden, which was associated with suppression of type 17 immunity and attenuation of antimicrobial peptide expression. Enhanced susceptibility to S. aureus coinfection ceased at day 14 of influenza infection, when influenza-associated type I IFN levels had returned to baseline levels, further suggesting a key role for type I IFN in coinfection pathogenesis. These findings further implicate type I IFN-associated suppression of type 17 immunity and antimicrobial peptide production as a conserved mechanism for enhanced susceptibility to both gram-positive and gram-negative bacterial coinfection during influenza infection.

Functional metamirrors using bianisotropic elements.

Conventional mirrors obey the simple reflection law that a plane wave is reflected as a plane wave, at the same angle. To engineer spatial distributions of fields reflected from a mirror, one can either shape the reflector or position some phase-correcting elements on top of a mirror surface. Here we show, both theoretically and experimentally, that full-power reflection with general control over the reflected wave phase is possible with a single-layer array of deeply subwavelength inclusions. These proposed artificial surfaces, metamirrors, provide various functions of shaped or nonuniform reflectors without utilizing any mirror. This can be achieved only if the forward and backward scattering of the inclusions in the array can be engineered independently, and we prove that it is possible using electrically and magnetically polarizable inclusions. The proposed subwavelength inclusions possess desired reflecting properties at the operational frequency band, while at other frequencies the array is practically transparent. The metamirror concept leads to a variety of applications over the entire electromagnetic spectrum, such as optically transparent focusing antennas for satellites, multifrequency reflector antennas for radio astronomy, low-profile conformal antennas for telecommunications, and nanoreflectarray antennas for integrated optics.

D-Amino acids incorporation in the frog skin-derived peptide esculentin-1a(1-21)NH2 is beneficial for its multiple functions.

Naturally occurring antimicrobial peptides (AMPs) represent promising future antibiotics. We have previously isolated esculentin-1a(1-21)NH2, a short peptide derived from the frog skin AMP esculentin-1a, with a potent anti-Pseudomonal activity. Here, we investigated additional functions of the peptide and properties responsible for these activities. For that purpose, we synthesized the peptide, as well as its structurally altered analog containing two D-amino acids. The peptides were then biophysically and biologically investigated for their cytotoxicity and immunomodulating activities. The data revealed that compared to the wild-type, the diastereomer: (1) is significantly less toxic towards mammalian cells, in agreement with its lower α-helical structure, as determined by circular dichroism spectroscopy; (2) is more effective against the biofilm form of Pseudomonas aeruginosa (responsible for lung infections in cystic fibrosis sufferers), while maintaining a high activity against the free-living form of this important pathogen; (3) is more stable in serum; (4) has a higher activity in promoting migration of lung epithelial cells, and presumably in healing damaged lung tissue, and (5) disaggregates and detoxifies the bacterial lipopolysaccharide (LPS), albeit less than the wild-type. Light scattering studies revealed a correlation between anti-LPS activity and the ability to disaggregate the LPS. Besides shedding light on the multifunction properties of esculentin-1a(1-21)NH2, the D-amino acid containing isomer may serve as an attractive template for the development of new anti-Pseudomonal compounds with additional beneficial properties. Furthermore, together with other studies, incorporation of D-amino acids may serve as a general approach to optimize the future design of new AMPs.