Novel Host Recognition Mechanism of the K1 Capsule-Specific Phage of Escherichia coli: Capsular Polysaccharide as the First Receptor and Lipopolysaccharide as the Secondary Receptor.

K1 capsule-specific phages of Escherichia coli have been reported in recent years, but the molecular mechanism involved in host recognition of these phages remains unknown. In this study, the interactions between PNJ1809-36, a new K1-specific phage, and its host bacterium, E. coli DE058, were investigated. A transposon mutation library was used to screen for receptor-related genes. Gene deletion, lysis curve determination, plaque formation test, adsorption assay, and inhibition assay of phage by lipopolysaccharide (LPS) showed that capsular polysaccharide (CPS) was the first receptor for the initial adsorption of PNJ1809-36 to E. coli DE058 and that LPS was a secondary receptor for the irreversible binding of the phage. The penultimate galactose in the outer core was identified as the specific binding region on LPS. Through antibody blocking assay, fluorescence labeling and high-performance gel permeation chromatography, the tail protein ORF261 of phage PNJ1809-36 was identified as the receptor-binding protein on CPS. Given these findings, we propose a model for the recognition process of phage PNJ1809-36 on E. coli DE058: the phage PNJ1809-36 tail protein ORF261 recognizes and adsorbs to the K1 capsule, and then the K1 capsule is partially degraded, exposing the active site of LPS which is recognized by phage PNJ1809-36. This model provides insight into the molecular mechanisms between K1-specific phages and their host bacteria. IMPORTANCE It has been speculated that CPS is the main receptor of K1-specific phages belonging to Siphoviridae. In recent years, a new type of K1-specific phage belonging to Myoviridae has been reported, but its host recognition mechanisms remain unknown. Here, we studied the interactions between PNJ1809-36, a new type of K1 phage, and its host bacterium, E. coli DE058. Our research showed that the phage initially adsorbed to the K1 capsule mediated by ORF261 and then bound to the penultimate galactose of LPS to begin the infection process.

Psychometric properties of the MOBITEC-GP mobile application for real-life mobility assessment in older adults.

Aim of this study was to test the reliability and validity of the life-space measures and walking speed delivered by the MOBITEC-GP app. Participants underwent several supervised walking speed assessments as well as a 1-week life-space assessment during two assessment sessions 9 days apart. Fifty-seven older adults (47.4% male, mean age= 75.3 (±5.9) years) were included in the study. The MOBITEC-GP app showed moderate to excellent test-retest reliability (ICCs between 0.584 and 0.920) and validity (ICCs between 0.468 and 0.950) of walking speed measurements of 50 meters and above and of most 1-week life-space parameters, including life-space area, time spent out-of-home, and action range. The MOBITEC-GP app for Android is a reliable and valid tool for the assessment of real-life walking speed (at distances of 50 metres and above) and life-space parameters of older adults. Future studies should look into technical issues more systematically in order to avoid invalid measurements.

Microencapsulated phages show prolonged stability in gastrointestinal environments and high therapeutic efficiency to treat Escherichia coli O157:H7 infection.

Escherichia coli (E. coli) O157:H7 bacterial infection causes severe disease in mammals and results in substantial economic losses worldwide. Due to the development of antibiotic resistance, bacteriophage (phage) therapy has become an alternative to control O157:H7 infection. However, the therapeutic effects of phages are frequently disappointing because of their low resistance to the gastrointestinal environment. In this study, to improve the stability of phages in the gastrointestinal tract, E. coli O157:H7 phages were microencapsulated and their in vitro stability and in vivo therapeutic efficiency were investigated. The results showed that compared to free phages, the resistance of microencapsulated phages to simulated gastric fluid and bile salts significantly increased. The microencapsulated phages were efficiently released into simulated intestinal fluid, leading to a better therapeutic effect in rats infected with E. coli O157:H7 compared to the effects of the free phages. In addition, the microencapsulated phages were more stable during storage than the free phages, showing how phage microencapsulation can play an essential role in phage therapy.

MOBIlity assessment with modern TEChnology in older patients' real-life by the General Practitioner: the MOBITEC-GP study protocol.

BACKGROUND: Mobility limitations in older adults are associated with poor clinical outcomes including higher mortality and disability rates. A decline in mobility (including physical function and life-space) is detectable and should be discovered as early as possible, as it can still be stabilized or even reversed in early stages by targeted interventions. General practitioners (GPs) would be in the ideal position to monitor the mobility of their older patients. However, easy-to-use and valid instruments for GPs to conduct mobility assessment in the real-life practice setting are missing. Modern technologies such as the global positioning system (GPS) and inertial measurement units (IMUs) - nowadays embedded in every smartphone - could facilitate monitoring of different aspects of mobility in the GP's practice. METHODS: This project's aim is to provide GPs with a novel smartphone application that allows them to quantify their older patients' mobility. The project consists of three parts: development of the GPS- and IMU-based application, evaluation of its validity and reliability (Study 1), and evaluation of its applicability and acceptance (Study 2). In Study 1, participants (target N = 72, aged 65+, ≥2 chronic diseases) will perform a battery of walking tests (varying distances; varying levels of standardization). Besides videotaping and timing (gold standard), a high-end GPS device, a medium-accuracy GPS/IMU logger and three different smartphone models will be used to determine mobility parameters such as gait speed. Furthermore, participants will wear the medium-accuracy GPS/IMU logger and a smartphone for a week to determine their life-space mobility. Participants will be re-assessed after 1 week. In Study 2, participants (target N = 60, aged 65+, ≥2 chronic diseases) will be instructed on how to use the application by themselves. Participants will perform mobility assessments independently at their own homes. Aggregated test results will also be presented to GPs. Acceptance of the application will be assessed among patients and GPs. The application will then be finalized and publicly released. DISCUSSION: If successful, the MOBITEC-GP application will offer health care providers the opportunity to follow their patients' mobility over time and to recognize impending needs (e.g. for targeted exercise) within pre-clinical stages of decline.

Assessing and Mapping of Road Surface Roughness based on GPS and Accelerometer Sensors on Bicycle-Mounted Smartphones.

The surface roughness of roads is an essential road characteristic. Due to the employed carrying platforms (which are often cars), existing measuring methods can only be used for motorable roads. Until now, there has been no effective method for measuring the surface roughness of un-motorable roads, such as pedestrian and bicycle lanes. This hinders many applications related to pedestrians, cyclists and wheelchair users. In recognizing these research gaps, this paper proposes a method for measuring the surface roughness of pedestrian and bicycle lanes based on Global Positioning System (GPS) and accelerometer sensors on bicycle-mounted smartphones. We focus on the International Roughness Index (IRI), as it is the most widely used index for measuring road surface roughness. Specifically, we analyzed a computing model of road surface roughness, derived its parameters with GPS and accelerometers on bicycle-mounted smartphones, and proposed an algorithm to recognize potholes/humps on roads. As a proof of concept, we implemented the proposed method in a mobile application. Three experiments were designed to evaluate the proposed method. The results of the experiments show that the IRI values measured by the proposed method were strongly and positively correlated with those measured by professional instruments. Meanwhile, the proposed algorithm was able to recognize the potholes/humps that the bicycle passed. The proposed method is useful for measuring the surface roughness of roads that are not accessible for professional instruments, such as pedestrian and cycle lanes. This work enables us to further study the feasibility of crowdsourcing road surface roughness with bicycle-mounted smartphones.

Enrichment of OpenStreetMap Data Completeness with Sidewalk Geometries Using Data Mining Techniques.

Tailored routing and navigation services utilized by wheelchair users require certain information about sidewalk geometries and their attributes to execute efficiently. Except some minor regions/cities, such detailed information is not present in current versions of crowdsourced mapping databases including OpenStreetMap. CAP4Access European project aimed to use (and enrich) OpenStreetMap for making it fit to the purpose of wheelchair routing. In this respect, this study presents a modified methodology based on data mining techniques for constructing sidewalk geometries using multiple GPS traces collected by wheelchair users during an urban travel experiment. The derived sidewalk geometries can be used to enrich OpenStreetMap to support wheelchair routing. The proposed method was applied to a case study in Heidelberg, Germany. The constructed sidewalk geometries were compared to an official reference dataset ("ground truth dataset"). The case study shows that the constructed sidewalk network overlays with 96% of the official reference dataset. Furthermore, in terms of positional accuracy, a low Root Mean Square Error (RMSE) value (0.93 m) is achieved. The article presents our discussion on the results as well as the conclusion and future research directions.

Vegetation dieback in the Mississippi River Delta triggered by acute drought and chronic relative sea-level rise.

Vegetation dieback and recovery may be dependent on the interplay between infrequent acute disturbances and underlying chronic stresses. Coastal wetlands are vulnerable to the chronic stress of sea-level rise, which may affect their susceptibility to acute disturbance events. Here, we show that a large-scale vegetation dieback in the Mississippi River Delta was precipitated by salt-water incursion during an extreme drought in the summer of 2012 and was most severe in areas exposed to greater flooding. Using 16 years of data (2007-2022) from a coastwide network of monitoring stations, we show that the impacts of the dieback lasted five years and that recovery was only partial in areas exposed to greater inundation. Dieback marshes experienced an increase in percent time flooded from 43% in 2007 to 75% in 2022 and a decline in vegetation cover and species richness over the same period. Thus, while drought-induced high salinities and soil saturation triggered a significant dieback event, the chronic increase in inundation is causing a longer-term decline in cover, more widespread losses, and reduced capacity to recover from acute stressors. Overall, our findings point to the importance of mitigating the underlying stresses to foster resilience to both acute and persistent causes of vegetation loss.

The eNOS-induced leonurine's new role in improving the survival of random skin flap.

In reconstructive and plastic surgery, random skin flaps are commonly utilized to treat skin abnormalities produced by a variety of factors. Flap delay procedure is commonly used to reduce flap necrosis. Due to the limitations of various conditions, the traditional surgical improvement can't effectively alleviate the skin flap necrosis. And leonurine (Leo) has antioxidant and anti-inflammatory effects. In this study, we researched the mechanism underlying the influences of varied Leo concentrations on the survival rate of random skin flaps. Our results showed that after Leo treatment, tissue edema and necrosis of the flap were significantly reduced, while angiogenesis and flap perfusion were significantly increased. Through immunohistochemistry and Western blot, we proved that Leo treatment can upregulate the level of angiogenesis, while Leo treatment significantly reduced the expression levels of oxidative stress, apoptosis and inflammation. As a result, it can significantly improve the overall viability of the random skin flaps through the increase of angiogenesis, restriction of inflammation, attenuation of oxidative stress, and reduction of apoptosis. And this protective function was inhibited by LY294002 (a broad-spectrum inhibitor of PI3K) and L-NAME (NG- nitro-L-arginine methyl ester, a non-selective NOS inhibitor). All in all, Leo is an effective drug that can activate the eNOS via the PI3K/Akt pathway. By encouraging angiogenesis, preventing inflammation, minimizing oxidative stress, and lowering apoptosis, Leo can raise the survival rate of random skin flaps. The recommended concentration of Leo in this study was 30 mg/kg.

Extraintestinal Pathogenic Escherichia coli Utilizes Surface-Located Elongation Factor G to Acquire Iron from Holo-Transferrin.

Extraintestinal pathogenic Escherichia coli (ExPEC) can cause systemic infections in both humans and animals. As an essential nutrient, iron is strictly sequestered by the host. Circumventing iron sequestration is a determinant factor for ExPEC infection. However, the ExPEC iron acquisition mechanism, particularly the mechanism of transferrin (TF) acquisition, remains unclear. This study reports that iron-saturated holo-TF can be utilized by ExPEC to promote its growth in culture medium and survival in macrophages. ExPEC specifically bound to holo-TF instead of iron-free apo-TF via the surface located elongation factor G (EFG) in both culture medium and macrophages. As a moonlighting protein, EFG specifically bound holo-TF and also released iron in TF. These two functions were performed by different domains of EFG, in which the N-terminal domains were responsible for holo-TF binding and the C-terminal domains were responsible for iron release. The functions of EFG and its domains have also been further confirmed by surface-display vectors. The surface overexpression of EFG bound significantly more holo-TF in macrophages and significantly improved bacterial intracellular survival ability. Our findings reveal a novel iron acquisition mechanism involving EFG, which suggests novel research avenues into the molecular mechanism of ExPEC resistance to nutritional immunity. IMPORTANCE Extraintestinal pathogenic Escherichia coli (ExPEC) is an important pathogen causing systemic infections in humans and animals. The competition for iron between ExPEC and the host is a determinant for ExPEC to establish a successful infection. Here, we sought to elucidate the role of transferrin (TF) in the interaction between ExPEC and the host. Our results revealed that holo-TF could be utilized by ExPEC to enhance its growth in culture medium and survival in macrophages. Furthermore, the role of elongation factor G (EFG), a novel holo-TF-binding and TF-iron release protein, was confirmed in this study. Our work provides insights into the iron acquisition mechanism of ExPEC, deepens understanding of the interaction between holo-TF and pathogens, and broadens further researches into the molecular mechanism of ExPEC pathogenicity.

Hcp Proteins of the Type VI Secretion System Promote Avian Pathogenic E. coli DE205B (O2:K1) to Induce Meningitis in Rats.

Avian pathogenic Escherichia coli (APEC) is an important extra-intestinal pathogenic E. coli (ExPEC), which often causes systemic infection in poultry and causes great economic loss to the breeding industry. In addition, as a major source of human ExPEC infection, the potential zoonotic risk of APEC has been an ongoing concern. Previous studies have pointed out that APEC is a potential zoonotic pathogen, which has high homology with human pathogenic E. coli such as uro-pathogenic E. coli (UPEC) and neonatal meningitis E. coli (NMEC), shares multiple virulence factors and can cause mammalian diseases. Previous studies have reported that O18 and O78 could cause different degrees of meningitis in neonatal rats, and different serotypes had different degrees of zoonotic risk. Here, we compared APEC DE205B (O2:K1) with NMEC RS218 (O18:K1:H7) by phylogenetic analysis and virulence gene identification to analyze the potential risk of DE205B in zoonotic diseases. We found that DE205B possessed a variety of virulence factors associated with meningitis and, through phylogenetic analysis, had high homology with RS218. DE205B could colonize the cerebrospinal fluid (CSF) of rats, and cause meningitis and nerve damage. Symptoms and pathological changes in the brain were similar to RS218. In addition, we found that DE205B had a complete T6SS, of which Hcp protein was its important structural protein. Hcp1 induced cytoskeleton rearrangement in human brain microvascular endothelial cells (HBMECs), and Hcp2 was mainly involved in the invasion of DE205B in vitro. In the meningitis model of rats, deletion of hcp2 gene reduced survival in the blood and the brain invasiveness of DE205B. Compared with WT group, Δhcp2 group induced lower inflammation and neutrophils infiltration in brain tissue, alleviating the process of meningitis. Together, these results suggested that APEC DE205B had close genetic similarities to NMEC RS218, and a similar mechanism in causing meningitis and being a risk for zoonosis. This APEC serotype provided a basis for zoonotic research.

The Key Factors in Physical Activity Type Detection Using Real-Life Data: A Systematic Review.

Background: Physical activity (PA) is paramount for human health and well-being. However, there is a lack of information regarding the types of PA and the way they can exert an influence on functional and mental health as well as quality of life. Studies have measured and classified PA type in controlled conditions, but only provided limited insight into the validity of classifiers under real-life conditions. The advantage of utilizing the type dimension and the significance of real-life study designs for PA monitoring brought us to conduct a systematic literature review on PA type detection (PATD) under real-life conditions focused on three main criteria: methods for detecting PA types, using accelerometer data collected by portable devices, and real-life settings. Method: The search of the databases, Web of Science, Scopus, PsycINFO, and PubMed, identified 1,170 publications. After screening of titles, abstracts and full texts using the above selection criteria, 21 publications were included in this review. Results: This review is organized according to the three key elements constituting the PATD process using real-life datasets, including data collection, preprocessing, and PATD methods. Recommendations regarding these key elements are proposed, particularly regarding two important PA classes, i.e., posture and motion activities. Existing studies generally reported high to near-perfect classification accuracies. However, the data collection protocols and performance reporting schemes used varied significantly between studies, hindering a transparent performance comparison across methods. Conclusion: Generally, considerably less studies focused on PA types, compared to other measures of PA assessment, such as PA intensity, and even less focused on real-life settings. To reliably differentiate the basic postures and motion activities in real life, two 3D accelerometers (thigh and hip) sampling at 20 Hz were found to provide the minimal sensor configuration. Decision trees are the most common classifier used in practical applications with real-life data. Despite the significant progress made over the past year in assessing PA in real-life settings, it remains difficult, if not impossible, to compare the performance of the various proposed methods. Thus, there is an urgent need for labeled, fully documented, and openly available reference datasets including a common evaluation framework.