New Evolutionary Insights into RpoA: A Novel Quorum Sensing Reprograming Factor in Pseudomonas aeruginosa.

Quorum-sensing (QS) coordinates the expression of virulence factors in Pseudomonas aeruginosa, an opportunistic pathogen known for causing severe infections in immunocompromised patients. QS has a master regulator, the lasR gene, but in clinical settings, P. aeruginosa isolates have been found that are QS-active but LasR-null. In this study, we developed an experimental evolutionary approach to identify additional QS-reprogramming determinants. We began the study with a LasR-null mutant with an extra copy of mexT, a transcriptional regulator gene that is known to be able to reprogram QS in laboratory LasR-null strains. In this strain, spontaneous single mexT mutations are expected to have no or little phenotypic consequences. Using this novel method, which we have named "targeted gene duplication followed by mutant screening", we identified QS-active revertants with mutations in genes other than mexT. One QS-active revertant had a point mutation in rpoA, a gene encoding the α-subunit of RNA polymerase. QS activation in this mutant was found to be associated with the downregulated expression of mexEF-oprN efflux pump genes. Our study therefore uncovers a new functional role for RpoA in regulating QS activity. Our results indicate that a RpoA-dependent regulatory circuit controlling the expression of the mexEF-oprN operon is critical for QS-reprogramming. In conclusion, our study reports on the identification of non-MexT proteins associated with QS-reprogramming in a laboratory strain, shedding light on possible QS activation mechanisms in clinical P. aeruginosa isolates.

Requirement for PINCH in skeletal myoblast differentiation.

PINCH, an adaptor of focal adhesion complex, plays essential roles in multiple cellular processes and organogenesis. Here, we ablated PINCH1 or both of PINCH1 and PINCH2 in skeletal muscle progenitors using MyoD-Cre. Double ablation of PINCH1 and PINCH2 resulted in early postnatal lethality with reduced size of skeletal muscles and detachment of diaphragm muscles from the body wall. PINCH mutant myofibers failed to undergo multinucleation and exhibited disrupted sarcomere structures. The mutant myoblasts in culture were able to adhere to newly formed myotubes but impeded in cell fusion and subsequent sarcomere genesis and cytoskeleton organization. Consistent with this, expression of integrin ß1 and some cytoskeleton proteins and phosphorylation of ERK and AKT were significantly reduced in PINCH mutants. However, N-cadherin was correctly expressed at cell adhesion sites in PINCH mutant cells, suggesting that PINCH may play a direct role in myoblast fusion. Expression of MRF4, the most highly expressed myogenic factor at late stages of myogenesis, was abolished in PINCH mutants that could contribute to observed phenotypes. In addition, mice with PINCH1 being ablated in myogenic progenitors exhibited only mild centronuclear myopathic changes, suggesting a compensatory role of PINCH2 in myogenic differentiation. Our results revealed a critical role of PINCH proteins in myogenic differentiation.

Hydrophobic Composite Foams with Asymmetric Gradient Sandwich Structure for Excellent Electromagnetic Interference Shielding and Photothermal Response.

The evolution of contemporary electronics urgently requires the use of versatile electromagnetic interference (EMI) shielding materials in complex environments. Interlayer polydimethylsiloxane (PDMS)/Fe3O4@multiwalled carbon nanotubes (MWCNTs) foams were prepared by a simple physical foaming method with excellent flexibility and electromagnetic wave absorption. The bottom nickel aramid paper (NiP) layer creates a dense conductive network by chemical plating technology, which ensures excellent EMI effectiveness. The upper carbon black (CB)/Fe3O4 layer further improves the absorption performance via conductive loss and magnetic loss. With the effective layout of the impedance matching layer, absorbing layer, and conductive shielding layer, the CB/Fe3O4-PDMS/Fe3O4@MWCNTs-NiP composite material achieves an EMI shielding effectiveness (EMI SE) of 61.7 dB and an absorption coefficient of 0.58 at X-band. In addition, the composite foam provides photothermal conversion and hydrophobicity due to the effective stacking of PDMS and CB/Fe3O4. Thus, the multifunctional composite foam presents a broad range of possible applications, benefiting EMI shielding as well as other specific areas.

Effects of Nitrogen Supply on Induced Defense in Maize (Zea mays) against Fall Armyworm (Spodoptera frugiperda).

How nitrogen (N) supply affects the induced defense of plants remains poorly understood. Here, we investigated the impacts of N supply on the defense induced in maize (Zea mays) against the fall armyworm (Spodoptera frugiperda). In the absence of herbivore attack or exogenous jasmonic acid (JA) application, N supply increased plant biomass and enhanced maize nutrient (soluble sugar and amino acid) contents and leaf area fed by S. frugiperda (the feeding leaf area of S. frugiperda larvae in maize supplemented with 52.2 and 156.6 mg/kg of N was 4.08 and 3.83 times that of the control, respectively). When coupled with herbivore attack or JA application, maize supplemented with 52.2 mg/kg of N showed an increased susceptibility to pests, while the maize supplemented with 156.6 mg/kg of N showed an improved defense against pests. The changes in the levels of nutrients, and the emissions of volatile organic compounds (VOCs) caused by N supply could explain the above opposite induced defense in maize. Compared with herbivore attack treatment, JA application enhanced the insect resistance in maize supplemented with 156.6 mg/kg of N more intensely, mainly reflecting a smaller feeding leaf area, which was due to indole emission and two upregulated defensive genes, MPI (maize proteinase inhibitor) and PAL (phenylalanine ammonia-lyase). Hence, the optimal N level and appropriate JA application can enhance plant-induced defense against pests.

Expression of ILK in renal stroma is essential for multiple aspects of renal development.

Kidney development involves reciprocal and inductive interactions between the ureteric bud (UB) and surrounding metanephric mesenchyme. Signals from renal stromal lineages are essential for differentiation and patterning of renal epithelial and mesenchymal cell types and renal vasculogenesis; however, underlying mechanisms remain not fully understood. Integrin-linked kinase (ILK), a key component of integrin signaling pathway, plays an important role in kidney development. However, the role of ILK in renal stroma remains unknown. Here, we ablated ILK in renal stromal lineages using a platelet-derived growth factor receptor B ( Pdgfrb) -Cre mouse line, and the resulting Ilk mutant mice presented postnatal growth retardation and died within 3 wk of age with severe renal developmental defects. Pdgfrb-Cre;Ilk mutant kidneys exhibited a significant decrease in UB branching and disrupted collecting duct formation. From E16.5 onward, renal interstitium was disorganized, forming medullary interstitial pseudocysts. Pdgfrb-Cre;Ilk mutants exhibited renal vasculature mispatterning and impaired glomerular vascular differentiation. Impaired glial cell-derived neurotrophic factor/Ret and bone morphogenetic protein 7 signaling pathways were observed in Pdgfrb-Cre;Ilk mutant kidneys. Furthermore, phosphoproteomic and Western blot analyses revealed a significant dysregulation of a number of key signaling pathways required for kidney morphogenesis, including PI3K/AKT and MAPK/ERK in Pdgfrb-Cre;Ilk mutants. Our results revealed a critical requirement for ILK in renal-stromal and vascular development, as well as a noncell autonomous role of ILK in UB branching morphogenesis.

Efficient directional excitation of surface plasmons by a single-element nanoantenna.

Directional light scattering is important in basic research and real applications. This area has been successfully downscaled to wavelength and subwavelength scales with the development of optical antennas, especially single-element nanoantennas. Here, by adding an auxiliary resonant structure to a single-element plasmonic nanoantenna, we show that the highly efficient lowest-order antenna mode can be effectively transferred into inactive higher-order modes. On the basis of this mode conversion, scattered optical fields can be well manipulated by utilizing the interference between different antenna modes. Both broadband directional excitation of surface plasmon polaritons (SPPs) and inversion of SPP launching direction at different wavelengths are experimentally demonstrated as typical examples. The proposed strategy based on mode conversion and mode interference provides new opportunities for the design of nanoscale optical devices, especially directional nanoantennas.

Discovery andw biological evaluation of novel 6,7-disubstituted-4-(2-fluorophenoxy)quinoline derivatives possessing 1,2,3-triazole-4-carboxamide moiety as c-Met kinase inhibitors.

A series of 6,7-disubstituted-4-(2-fluorophenoxy)quinoline derivatives possessing 1,2,3-triazole-4-carboxamide moiety were designed, synthesized and evaluated for their in vitro biological activities against c-Met kinase and five typical cancer cell lines (A549, H460, HT-29, MKN-45 and U87MG). Most compounds showed moderate to excellent antiproliferative activity. In this study, a promising compound 34, with a c-Met IC50 value of 1.04nM, was identified as a multitargeted receptor tyrosine kinase inhibitor. The SAR analyses indicated that compounds with halogen group, especially fluoro group, at 4-position on the phenyl ring (moiety B) have potent antitumor activity, and methylation on the 5-atom linker played an important role in the c-Met enzymatic activity.

Using text mining and multilevel association rules to process and analyze incident reports in China.

Incident investigation reports provide information on defects related to the system safety and indications for improvements. Currently, the analysis of these reports relies heavily on expert' experience. The foreseeable work-load and lack of understanding about the importance of near misses have created a situation where severe accidents are rigorously investigated, and minor incidents are often omitted. Consequently, incident reports have not been fully analyzed to provide sufficient solutions. The aim of this research is to propose a framework that uses text mining and multilevel association rules to efficiently structure Chinese incident reports and identify important incident patterns, providing an analysis of trends, rectification strategies, and guidance for safety management. A case study of a construction company in China was conducted using two years of incident data dated 2018-2019, including accidents and near misses. To identify incident elements, a pattern extraction workflow involving TextRank, and domain pertinence was devised based on the linguistic and writing styles of Chinese reports. A concept hierarchy was applied to determine the taxonomic relationships within the risk factors. Multilevel association rule mining was adopted and proven to deliver more comprehensive pattern indications. Comparative and cross-analysis of patterns in different time periods revealed the severity and temporal features of incidents as well as the effectiveness of preventive and precautionary measures. The results also highlight the importance of learning from near miss events. Decision makers can formulate countermeasures and management policies based on these results to improve safety performance.

Four-Way Crossover Phase 1 Thorough QT Study to Assess Cardiac Safety of Domperidone and Consideration for Design and Conduct of QT/QTc Study in China.

This study aimed to evaluate the QT prolongation potential of domperidone in healthy Chinese participants and explore the possibility of a thorough QT (TQT) study in China with a smaller sample size using concentration-QT (C-QT) modeling. Part 1 was a randomized, placebo- and positive-controlled, multiple-dose, 4-way crossover TQT study in healthy Chinese participants; 44 participants were randomized to either domperidone 10/20 mg or placebo 3 times daily, on days 1 to 3, followed by a single dose of either 10/20 mg domperidone/domperidone-placebo/domperidone-placebo plus 400 mg of moxifloxacin, on day 4. Twelve-lead electrocardiograms were recorded in triplicate at predefined time points with pharmacokinetic sampling. The results were that change from baseline in QT interval corrected for heart rate (QTc) using the Fridericia formula (QTcF) between domperidone and placebo was 1.3 milliseconds and 2.7 milliseconds for 10 and 20 mg 3 times daily, and upper limits of 2-sided 90%CI for all time points were below regulatory threshold of 10 milliseconds. In part 2, resampling analysis using C-QT modeling for moxifloxacin showed false-negative rates of <5% with sample sizes ≥6. We could conclude that no clinically relevant effect on corrected QT interval or new safety signals was observed with domperidone. A dedicated TQT study with C-QT modeling could assess drug effects on QT/corrected QT intervals for novel drug development in China.

Elevated Levels of Circulating Biomarkers Related to Leaky Gut Syndrome and Bacterial Translocation Are Associated With Graves' Disease.

Background: A growing number of studies have found dysbiosis of the intestinal microbiota in patients with Graves' disease (GD). The intestinal epithelial barrier serves as the first line of defense, protecting the immune system from excessive stimulation of microbiota and toxins. Most autoimmune diseases are associated with a gut barrier dysfunction (leaky gut) which allows bacterial translocation. However, to date, potential correlations between intestinal barrier dysfunction and GD have not been explored. Methods: Serum lipopolysaccharide (LPS), intestinal fatty acid-binding protein (I-FABP), zonulin, D-lactate, and diamine oxidase (DAO) were measured to assess barrier integrity in 91 patients with GD (61 initial GD and 30 euthyroid GD) and 44 healthy controls. The quality of life (QOL) of patients with GD was assessed using the thyroid-specific patient-reported outcome (ThyPRO-39) questionnaire. Results: The serum levels of LPS, I-FABP, zonulin, and D-lactate were significantly higher in patients with initial GD than in healthy controls. Logistic regression analysis revealed that zonulin and D-lactate were independently associated with risk for GD and circulating zonulin could effectively distinguish patients with initial GD from healthy controls. Correlation analyses showed that I-FABP, LPS, and D-lactate were positively associated with FT4 and negatively associated with TSH. In addition, circulating LPS, zonulin, and D-lactate levels were all independent predictors of TRAb levels. Moreover, higher circulating LPS levels in patients with GD were associated with more severe hyperthyroidism (higher concentrations of FT3, FT4, and TRAb and lower TSH concentrations) and worse scores of hyperthyroid and eye symptoms. Conclusion: Patients with initial GD show a disrupted intestinal barrier, characterized by elevated levels of leaky gut biomarkers. Increased intestinal permeability and bacterial translocation were associated with TRAb levels and hyperthyroidism in GD. Further research is required to elucidate the underlying mechanisms.

A survey of variable selection methods in two Chinese epidemiology journals.

BACKGROUND: Although much has been written on developing better procedures for variable selection, there is little research on how it is practiced in actual studies. This review surveys the variable selection methods reported in two high-ranking Chinese epidemiology journals. METHODS: Articles published in 2004, 2006, and 2008 in the Chinese Journal of Epidemiology and the Chinese Journal of Preventive Medicine were reviewed. Five categories of methods were identified whereby variables were selected using: A--bivariate analyses; B--multivariable analysis; e.g. stepwise or individual significance testing of model coefficients; C--first bivariate analyses, followed by multivariable analysis; D--bivariate analyses or multivariable analysis; and E--other criteria like prior knowledge or personal judgment. RESULTS: Among the 287 articles that reported using variable selection methods, 6%, 26%, 30%, 21%, and 17% were in categories A through E, respectively. One hundred sixty-three studies selected variables using bivariate analyses, 80% (130/163) via multiple significance testing at the 5% alpha-level. Of the 219 multivariable analyses, 97 (44%) used stepwise procedures, 89 (41%) tested individual regression coefficients, but 33 (15%) did not mention how variables were selected. Sixty percent (58/97) of the stepwise routines also did not specify the algorithm and/or significance levels. CONCLUSIONS: The variable selection methods reported in the two journals were limited in variety, and details were often missing. Many studies still relied on problematic techniques like stepwise procedures and/or multiple testing of bivariate associations at the 0.05 alpha-level. These deficiencies should be rectified to safeguard the scientific validity of articles published in Chinese epidemiology journals.

Defect development in a three-dimensional reaction-diffusion system with gradient.

The formation and development of spiral defects is one of the major causes of order-disorder transitions in spatiotemporal patterns. In this paper, line defect formation and development in a three-dimensional reaction-diffusion system with gradients of control parameters in the third dimension is investigated. The system can be considered as diffusively coupled two-dimensional spatiotemporal patterns with dissimilarities. We observed that under certain conditions, as the gradients are varied, ordered and disordered spatiotemporal patterns appear alternately and line defects of various configurations form. This scenario is found to be in qualitative agreement with the experimental findings in the Belousov-Zhabotinsky reaction. We thus demonstrate that the line defect which was usually expected in two-dimensional complex oscillatory media can also be generated from the reconciliation between the coupled simple spiral waves with dissimilarity.

High-frequency ultrasonography and contrast-enhanced ultrasound for the evaluation of testicular capillary hemangioma: A case report.

RATIONALE: Testicular capillary hemangioma (TCH) is a rare benign tumor of the testis. To the authors' knowledge, there is currently only a few literatures describing the use of contrast-enhanced ultrasound (CEUS) to diagnose TCH. Accurate preoperative diagnosis of benign tumors can avoid orchiectomy. A case of TCH evaluated using high-frequency ultrasound and CEUS is presented. PATIENT CONCERNS: A 21-year-old male presented with a right testicular mass during a routine physical examination, and was admitted to the authors' hospital for definitive diagnosis and treatment. DIAGNOSES: Combined gray-scale, color Doppler ultrasonography, and CEUS did not exclude the possibility that the right testicular lesion may be a benign tumor. Combined with morphological and immunohistochemical staining results, a pathological diagnosis of TCH was considered. INTERVENTIONS: The patient underwent right orchiectomy under general anesthesia, which proceeded smoothly. OUTCOMES: At the 12-month follow-up, the patient was completely asymptomatic and resumed all daily activities. LESSONS: TCH is a rare benign tumor and lacks extensive previous data in imaging findings. If TCH can be diagnosed accurately before surgery, excessive or inappropriate treatment of benign lesions can be minimized, which will be beneficial to the physical and psychological health of patients.

Collaborative ISL1/GATA3 interaction in controlling neuroblastoma oncogenic pathways overlapping with but distinct from MYCN.

Background: Transcription factor ISL1 plays a critical role in sympathetic neurogenesis. Expression of ISL1 has been associated with neuroblastoma, a pediatric tumor derived from sympatho-adrenal progenitors, however the role of ISL1 in neuroblastoma remains unexplored. Method: Here, we knocked down ISL1 (KD) in SH-SY5Y neuroblastoma cells and performed RNA-seq and ISL1 ChIP-seq analyses. Results: Analyses of these data revealed that ISL1 acts upstream of multiple oncogenic genes and pathways essential for neuroblastoma proliferation and differentiation, including LMO1 and LIN28B. ISL1 promotes expression of a number of cell cycle associated genes, but represses differentiation associated genes including RA receptors and the downstream target genes EPAS1 and CDKN1A. Consequently, Knockdown of ISL1 inhibits neuroblastoma cell proliferation and migration in vitro and impedes tumor growth in vivo, and enhances neuronal differentiation by RA treatment. Furthermore, genome-wide mapping revealed a substantial co-occupancy of binding regions by ISL1 and GATA3, and ISL1 physically interacts with GATA3, and together they synergistically regulate the aforementioned oncogenic pathways. In addition, analyses of the roles of ISL1 and MYCN in MYCN-amplified and MYCN non-amplified neuroblastoma cells revealed an epistatic relationship between ISL1 and MYCN. ISL1 and MYCN function in parallel to regulate common yet distinct oncogenic pathways in neuroblastoma. Conclusion: Our study has demonstrated that ISL1 plays an essential role in neuroblastoma regulatory networks and may serve as a potential therapeutic target in neuroblastoma.

Influence of hydrogel mechanical properties and mesh size on vocal fold fibroblast extracellular matrix production and phenotype.

Current clinical management of vocal fold (VF) scarring produces inconsistent and often suboptimal results. Researchers are investigating a number of alternative treatments for VF lamina propria (LP) scarring, including designer implant materials for functional LP regeneration. In the present study, we investigate the effects of the initial scaffold elastic modulus and mesh size on encapsulated VF fibroblast (VFF) extracellular matrix (ECM) production toward rational scaffold design. Poly(ethylene glycol) diacrylate (PEGDA) hydrogels were selected for this study since their material properties, including mechanical properties, mesh size, degradation rate and bioactivity, can be tightly controlled and systematically modified. Porcine VFF were encapsulated in four PEGDA hydrogels with degradation half lives of approximately 25 days, but with initial elastic compressive moduli and mesh sizes ranging from approximately 30 to 100kPa and from approximately 9 to 27nm, respectively. After 30 days of static culture, VFF ECM production and phenotype in each formulation was assessed biochemically and histologically. Sulfated glycosaminoglycan synthesis increased in similar degree with both increasing initial modulus and decreasing initial mesh size. In contrast, elastin production decreased with increasing initial modulus but increased with decreasing initial mesh size. Both collagen deposition and the induction of a myofibroblastic phenotype depended strongly on initial mesh size but appeared largely unaffected by variations in initial modulus. The present results indicate that scaffold mesh size warrants further investigation as a critical regulator of VFF ECM synthesis. Furthermore, this study validates a systematic and controlled approach for analyzing VFF response to scaffold properties, which should aid in rational scaffold selection/design.

Local wave grouping in a parameter-gradient system and its formation mechanism.

In a ferroin-catalyzed Belousov-Zhabotinsky (BZ) reaction-diffusion system with reagent concentration gradients, we observed in the experiment a type of spirals with local waves forming groups. Here, we propose an interpretation of the wave grouping phenomenon. The wave grouping mechanism can be well explained in terms of the cooperation of the excitability gradient and the Doppler effect induced by spiral tip's meandering. In the simulation based on three-dimensional reaction-diffusion system using Oregonator model, spiral patterns analogous to the experiment observation are well reproduced when the parameter gradient in the z direction is introduced.

Chemical turbulence and line defects induced by gradient effects in a three-dimensional reaction-diffusion system.

We report experimental observations of chemical turbulence and line defects in a three-dimensional (3-D) Belousov-Zhabotinsky (BZ) reaction-diffusion system. Transitions from spiral waves to 3-D chemical turbulence to line defects are observed. These transitions are caused by concentration gradients across the third dimension in the 3-D reaction medium, indicating the observed line defects have a 3-D structure. The line defects come out of the 3-D turbulent state, and become smooth with the increase of the control parameter. Simulation with the two-variable Oregonator model in the 3-D system reproduces similar line defects.

Systemic cat scratch disease.

Systemic cat scratch disease (CSD) is often associated with prolonged fever and microabscesses in the liver and/or spleen. We report a case of systemic CSD with hepatic, splenic and renal involvement in an aboriginal child in Taiwan. A previously healthy 9-year-old girl had an intermittent fever for about 17 days, and complained of abdominal pain, headache and weight loss. Abdominal computed tomography showed multiple tiny hypodense nodular lesions in the spleen and both kidneys. Laparotomy revealed multiple soft, whitish-tan lesions on the surface of the liver and spleen. Histopathologic examination of a biopsy specimen of the spleen showed necrotizing granulomatous inflammation with central necrosis surrounded by epithelioid cells and occasional Langhans' giant cells, strongly suggestive of Bartonella henselae infection. History revealed close contact with a cat. B. henselae DNA was detected by polymerase chain reaction in the tissue specimen, and the single antibody titer against B. henselae was greater than 1:2048. These results confirmed the diagnosis of visceral CSD caused by B. henselae. The patient's symptoms resolved after treatment with rifampin and tetracycline. This case illustrates the need for inclusion of systemic CSD in patients with fever of unknown origin and abdominal pain.

Efficient unidirectional launching of surface plasmons by a cascade asymmetric-groove structure.

Increasing the unidirectional launching efficiency of surface plasmon polaritons (SPPs) is crucial in plasmonics. Here, we demonstrate that this efficiency may be improved by cascading subwavelength unidirectional SPP launching units. A unidirectional SPP launching efficiency of at least 46% and an extinction ratio of 40 are experimentally demonstrated using a cascade asymmetric-groove structure. Meanwhile, the device is ultracompact, and has a lateral dimension of only 1.1 µm. The proposed structure also presents a broadband response and is easy to fabricate. This high-performance wavelength-scale unidirectional SPP launcher represents an important development in practical SPP sources.

Relation between the wave front and the tip movement of spirals.

The relation between the movement of spiral waves and their tip for a simple spiral is experimentally investigated in a spatial open reactor using the Belousov-Zhabotinsky reaction. In the quasi-two dimensional regime, our results agree with the combustion model proposed by Lázár et al. [Chaos 5, 443 (1995)] which states that the speed of spiral tip movement is proportional to the speed of spiral waves, c(tip) infinity c(front), when expressed in the radius of the spiral tip trajectory R and the spiral wavelength lambda, R infinity lambda. The measurement shows that the slope of the linear R-lambda curve is about 0.13, independent of the control parameter. This observation is also consistent with the combustion model.