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Chao Cao,1– 4 Obulkasim Memet,1– 3 Fuli Liu,1,2 Hanbing Hu,1– 3 Lin Zhang,1,2 Heng Jin,4 Yiqun Cao,4 Jian Zhou,3,5,6 Jie Shen1– 3 1Center of Emergency and Critical Medicine, Jinshan Hospital of Fudan University, Shanghai, People’s Republic of China; 2Research Center for Chemical Injury, Emergency and Critical Medicine of Fudan University, Shanghai, People’s Republic of China; 3Shanghai Medical College, Fudan University, Shanghai, People’s Republic of China; 4Emergency Department, Tianjin Medical University General Hospital, Tianjin, People’s Republic of China; 5Department of Pulmonary and Critical Care Medicine, Shanghai Respiratory Research Institute, Zhongshan Hospital of Fudan University, Shanghai, People’s Republic of China; 6Shanghai Institute of Infectious Disease and Biosecurity, Fudan University, Shanghai, People’s Republic of ChinaCorrespondence: Jie Shen, Center of Emergency and Critical Medicine, Jinshan Hospital of Fudan University, Shanghai, 201508, People’s Republic of China, Tel +86 2134189990, Email jshen_@fudan.edu.cnPurpose: Chemically induced acute lung injury (CALI) has become a serious health concern in our industrialized world, and abnormal functional alterations of immune cells crucially contribute to severe clinical symptoms. However, the cell heterogeneity and functional phenotypes of respiratory immune characteristics related to CALI remain unclear.Methods: We performed scRNA sequencing on bronchoalveolar lavage fluid (BALF) samples obtained from phosgene-induced CALI rat models and healthy controls. Transcriptional data and TotalSeq technology were used to confirm cell surface markers identifying immune cells in BALF. The landscape of immune cells could elucidate the metabolic remodeling mechanism involved in the progression of acute respiratory distress syndrome and cytokine storms. We used pseudotime inference to build macrophage trajectories and the corresponding model gene expression changes, and identified and characterized alveolar cells and immune subsets that may contribute to CALI pathophysiology based on gene expression profiles at single-cell resolution.Results: The immune environment of cells, including dendritic cells and specific macrophage subclusters, exhibited increased function during the early stage of pulmonary tissue damage. Nine different subpopulations were identified that perform multiple functional roles, including immune responses, pulmonary tissue repair, cellular metabolic cycle, and cholesterol metabolism. Additionally, we found that individual macrophage subpopulations dominate the cell-cell communication landscape. Moreover, pseudo-time trajectory analysis suggested that proliferating macrophage clusters exerted multiple functional roles.Conclusion: Our findings demonstrate that the bronchoalveolar immune microenvironment is a fundamental aspect of the immune response dynamics involved in the pathogenesis and recovery of CALI.Graphical Abstract: Keywords: single cell RNAseq, phosgene-inhalation, chemically induced acute lung injury, immune microenvironment, heterogeneity, phenotypes

Saccharina japonica is an ecologically and economically important kelp in cold-temperate region. When it is cultivated on a large scale in the temperate and even subtropical zones, heat stress is a frequent abiotic stress. This study is the first attempt to reveal the regulatory mechanism of the response to heat stress from the perspective of DNA methylation in S. japonica. We firstly obtained the characteristics of variation in the methylome under heat stress, and found that heat stress caused a slight increase in the overall methylation level and methylation rate, especially in the non-coding regions of the genome. Secondly, we found that methylation was probably one of factors affecting the expression of genes, and the methylation within the gene body was positively correlated with the gene expression (rho=0.0784). Moreover, it was found that among the differentially expressed genes regulated by methylation, many genes were proved to be related to heat stress response, such as HSP gene family, genes of antioxidant enzymes, genes related to proteasome-ubiquitination pathway and plant cell signaling pathways, etc. This study demonstrated that DNA methylation is involved in regulating the response to heat stress, laying a foundation for studying the acclimation and adaptation of S. japonica to heat stress from an epigenetic perspective.

Acute respiratory distress syndrome (ARDS) presents as a form of acute respiratory failure resulting from non-cardiogenic pulmonary edema due to excessive alveolocapillary permeability, which may be pulmonary or systemic in origin. In the last 3 years, the coronavirus disease 2019 pandemic has resulted in an increase in ARDS cases and highlighted the challenges associated with this syndrome, as well as the unacceptably high mortality rates and lack of effective treatments. Currently, clinical treatment remains primarily supportive, including mechanical ventilation and drug-based therapy. Mesenchymal stem cell (MSC) therapies are emerging as a promising intervention in patients with ARDS and have promising therapeutic effects and safety. The therapeutic mechanisms include modifying the immune response and assisting with tissue repair. This review provides an overview of the general properties of MSCs and outlines their role in mitigating lung injury and promoting tissue repair in ARDS. Finally, we summarize the current challenges in the study of translational MSC research and identify avenues by which the discipline may progress in the coming years.

Cobalt ferrite has problems such as poor impedance matching and high density, which results in unsatisfactory electromagnetic wave (EMW) absorption performance. In this study, the CoFe2O4@C core–shell structure composite was synthesized by a two-step hydrothermal method. X-ray diffraction, transmission electron microscopy, Fourier transform infrared spectroscopy, thermogravimetric analysis, and vector network analysis et al. were used to test the structure and EMW absorption properties of CoFe2O4@C composite. The results show that the reflection loss (RL) of the CoFe2O4@C composite reaches the maximum value of –25.66 dB at 13.92 GHz, and the effective absorbing band (EAB) is 4.59 GHz (11.20–15.79 GHz) when the carbon mass content is 6.01%. The RL and EAB of CoFe2O4@C composite are increased by 219.55% and 4.59 GHz respectively, and the density is decreased by 20.78% compared with the cobalt ferrite. Such enhanced EMW absorption properties of CoFe2O4@C composite are attributed to the attenuation caused by the strong natural resonance of the cobalt ferrite, moreover, the carbon coating layer adjusts the impedance matching of the composite, and the introduced dipole polarization and interface polarization can cause multiple Debye relaxation processes.

Cryptochrome/Photolyase Family (CPF) is a widely distributed and highly conserved gene family. The CPF includes photolyases that repair UV-induced DNA damage and blue-light receptors cryptochromes that are known for their photoreceptive functions in terrestrial plants. This study mainly emphasized the CPF members of the economically important brown alga Saccharina japonica, and traces the evolutionary relationship and distribution of this family in algae. Eight new CPF members were identified from the genomes of S. japonica and Undaria pinnatifida, and sixty CPF members from other different species were collected simultaneously for the phylogenetic analysis. Phylogenetic and transcriptional analyses provide evidence for the evolution and function of CPF genes in brown algae. According to the phylogenetic tree, the CPF family was divided into five branches. Photolyase homology domain (PHR) and a non-covalently bound flavin adenine dinucleotide (FAD) molecule are present in the CPF members of two brown algae S. japonica and U. pinnatifida. But they lack the typical C-terminal extension (CCT) characteristic of plant-like cryptochromes. The brown algae Ectocarpus siliculosus and U. pinnatifida have both animal-like CPF and plant-like CPF while only plant CPF members were identified in S. japonica. The results of RNA-seq and qPCR indicated that Sj Cryptochrome-DASH1 (CRY-DASH1) and Sj cyclobutane pyrimidine dimer (CPD) genes may not only play a role in light response but also play an important role in the gametophyte development of S. japonica. Protein interaction network of Sj CPF members showed that there are circadian clock protein Timeless homologous genes in S. japonica, which are located in the core position of the protein interaction network. This implied that Sj CPF members may transmit information by interacting with downstream Timeless genes. These results provide information for subsequent research on functional and regulatory pathways of CPFs in algae species.

Sargassum thunbergii is a brown macroalga endemic to the northwest Pacific. It plays important ecological roles in the structure and maintenance of coastal marine ecosystems. The bioactive compounds extracted from S. thunbergii have been extensively documented for potential use in anti-obesity, anti-inflammatory activity, anti-tumor, anti-oxidant and aquacultural drugs. The species is edible and contains relatively high levels of proteins, minerals and several types of amino acids. The present work compiles recently published literature on S. thunbergii, with particular focus on cultivation efforts in China, including the breeding of seedlings and cultivation at sea. A concise review of possible applications is given. Distribution, range shifts associated with past climate change, population genetic structure and connectivity, life history, reproduction and development are all detailed. The review provides important guidelines for future large-scale farming of S. thunbergii. This will help aquaculturalists (phyconomists) to meet the expected increases in demand by industrial users. It will also help to conserve natural populations which may be declining due to destructive harvesting and rapid ocean changes. 31971395; 2018YFD0900305; ZR2020MC033; UID/Multi/04326/2020; DL57/2016/CP1361/CT0035 info:eu-repo/semantics/publishedVersion

During tumorigenesis, tumor cells interact intimately with their surrounding cells (microenvironment) for their growth and progression. However, the roles of tumor microenvironment in tumor development and progression are not fully understood. Here, using an established benign tumor model in adult Drosophila intestines, we find that non-cell autonomous autophagy (NAA) is induced in tumor surrounding neighbor cells. Tumor growth can be significantly suppressed by genetic ablation of autophagy induction in tumor neighboring cells, indicating that tumor neighboring cells act as tumor microenvironment to promote tumor growth. Autophagy in tumor neighboring cells is induced downstream of elevated ROS and activated JNK signaling in tumor cells. Interestingly, we find that active transport of nutrients, such as amino acids, from tumor neighboring cells sustains tumor growth, and increasing nutrient availability could significantly restore tumor growth. Together, these data demonstrate that tumor cells take advantage of their surrounding normal neighbor cells as nutrient sources through NAA to meet their high metabolic demand for growth and progression. Thus we provide insights into our understanding of the mechanisms underlying the interaction between tumor cells and their microenvironment in tumor development.

Biosynthesis and replication, from A to Z Four nucleobases. adenine (A), cytosine (C), guanine (G), and thymine (T), are usually thought to be invariable in DNA. In bacterial viruses, however, each of the DNA bases have variations that help them to escape degradation by bacterial restriction enzymes. In the genome of cyanophage S-2L, A is completely replaced by diaminopurine (Z), which forms three hydrogen bonds with T and thus creates non–Watson-Crick base pairing in the DNA of this virus (see the Perspective by Grome and Isaacs). Zhou et al. and Sleiman et al. determined the biochemical pathway that produces Z, which revealed more Z genomes in viruses hosted in bacteria distributed widely in the environment and phylogeny. Pezo et al. identified a DNA polymerase that incorporates Z into DNA while rejecting A. These findings enrich our understanding of biodiversity and expand the genetic palette for synthetic biology. Science , this issue p. 512 , 516 , 520 ; see also p. 460

Leptospirosis, caused by pathogenic Leptospira species, has emerged as a widespread zoonotic disease worldwide. Macrophages mediate the elimination of pathogens through phagocytosis and cytokine production. Scavenger receptor A1 (SR-A1), one of the critical receptors mediating this process, plays a complicated role in innate immunity. However, the role of SR-A1 in the immune response against pathogenic Leptospira invasion is unknown. In the present study, we found that SR-A1 is an important nonopsonic phagocytic receptor on murine macrophages for Leptospira. However, intraperitoneal injection of leptospires into WT mice presented with more apparent jaundice, subcutaneous hemorrhaging, and higher bacteria burdens in blood and tissues than that of SR-A1-/- mice. Exacerbated cytokine and inflammatory mediator levels were also observed in WT mice and higher recruited macrophages in the liver than those of SR-A1-/- mice. Our findings collectively reveal that although beneficial in the uptake of Leptospira by macrophage, SR-A1 might be exploited by Leptospira to modulate inflammatory activation and increase the susceptibility of infection in the host. These results provide our new insights into the innate immune response during early infection by L. interrogans.

Mesenchymal stem cells (MSCs) have promising potential in the treatment of various diseases, such as the therapeutic effect of bone marrow-derived MSCs for phosgene-induced acute lung injury (P-ALI). However, MSC-related therapeutics are limited due to poor cell survival, requiring appropriate MSC delivery systems to maximise therapeutic capacity. Biomaterial RGD-hydrogel is a potential cell delivery vehicle as it can mimic the natural extracellular matrix and provide cell adhesion support. The application of RGD-hydrogel in the MSC treatment of respiratory diseases is scarce. This study reports that RGD-hydrogel has good biocompatibility and can increase the secretion of Angiopoietin-1, hepatocyte growth factor, epidermal growth factor, vascular endothelial cell growth factor, and interleukin-10 in vitro MSCs. The hydrogel-encapsulated MSCs could further alleviate P-ALI and show better cell survival in vivo. Overall, RGD-hydrogel could improve the MSC treatment of P-ALI by modulating cell survival and reparative activities. It is exciting to see more and more ways to unlock the therapeutic potential of MSCs.

In this paper, the solid waste desulfurization gypsum produced by coal-fired power plants was used as a raw material to prepare calcium sulfate whiskers with high application prospects. Calcium sulfate whiskers with uniform morphology and high aspect ratio can be prepared by hydrothermal method in sulfuric acid solution. A new process of desulfurization gypsum activated by high-energy grinding to reduce the reaction temperature and sulfuric acid concentration was developed. Through the comparison of product morphology, the best grinding time was determined to be 3.5 h. The mechanism of desulfurization gypsum through physical–chemical coupling to reduce energy consumption was clarified. The activation of desulfurization gypsum by grinding and the acidic environment provided by the sulfuric acid solution made the calcium sulfate solution reached rapid saturation and accelerated the nucleation rate. By calculating the conversion and crystallization rate of calcium sulfate whiskers, it was found that there were obvious “autocatalytic” kinetic characteristics during the crystallization process.

Copper ferrite modified carbon nanotubes (CuFe2O4/CNTs) were synthesized and employed as adsorbents/catalysts to assist electro-peroxone treatment of aqueous emerging pollutant fluconazole (FLC). Results suggest that the synthetic CuFe2O4/CNTs integrated the CNTs adsorption and ferrite magnetic abilities, showing a fast FLC removal and an excellent recyclability. The pseudo-second-order equation and the Langmuir isotherm model could well elaborate the adsorption of FLC on prepared catalysts. Compared to individual processes, electro-peroxone treatment exhibited a synergetic degradation of FLC. Approximately 89% FLC transformation was observed in the CuFe2O4/CNTs catalytic electro-peroxone process, 10% higher than that without catalysts, implying that CuFe2O4/CNTs could enhance the peroxone oxidation. Increasing of inlet gasphase ozone concentration (7.2–29.7 mg/L), catalysts dose (0.1–0.8 g/L), initial solution pH (3–11), and reaction temperature (10–30 °C) could improve the treatment efficiency, while initial FLC concentration (5–40 mg/L) and natural organic matter contents (humic acid, 0–20 mg/L) have opposite effects. The role of cathodic current density (10–60 mA/cm2) was complex, 40 mA/cm2 was observed as the optimal value. Furthermore, the degradation of FLC followed the hydroxyl radicals (HO ) oxidation mechanism. The production of HO was mainly via peroxone reactions, and the presence of CuFe2O4/CNTs could catalytic ozone and H2O2 decomposition, thereby promoting HO generation. Although the magnetic CuFe2O4/CNTs had not shown obvious microelectrode characteristics, it combined adsorption and catalytic peroxone oxidation, may serve as the catalyst for electro-peroxone process.

Saccharina japonica is a typical seaweed that has been domesticated and cultivated at a large scale for a long time worldwide. Compared to the wild populations, the Saccharina cultivars have significant greater high-temperature tolerance. However, the molecular mechanisms of high temperature tolerance are still unknown. This study characterized the protein expression profile under heat stress using the iTRAQ strategy integrated with transcriptome data to identify the candidate genes involved in the heat stress tolerance. A total of 104 and 107 proteins were identified as the differential expressed proteins (DEPs) in sporophyte and gametophyte, respectively. Only 14 DEPs identified both in sporophyte and gametophyte indicated that the two generations have different protein expression in response to the heat stress. The correlation between proteome and transcriptome profile under heat stress was very low, with only six genes were identified both on the transcription and protein levels. However, 23 pathways, including multiple cellular processes such as nitrogen and carbon metabolism, transcription, translation, posttranslational modification, antioxidant system etc., were identified. This study is the first exploration into the molecular mechanism of Saccharina heat tolerance on both transcriptomic and proteomic levels, which provides new insight into the regulation mechanism in response to heat stress in kelp.

Background Small-cell lung cancer (SCLC) has a poor prognosis since there is currently no effective therapy for commonly recurring disease. In our previous study, both primary and recurrent human tumors have been shown to express functional N-methyl-D-aspartate (NMDA) receptors, and blockade of these receptors with GluN1 and GluN2B antagonists decreased tumor cell viability in vitro, and growth of tumor xenografts in nu/nu mice. Materials and methods In this study, we examine the influence of the GluN2B antagonist ifenprodil and the channel-blocker antagonist memantine, on cell viability and growth of tumor xenografts of recurrent SCLC (rSCLC) in mice. Results Both antagonists significantly reduced cell viability and levels of components of the ERK1/2 pathway, increased apoptosis, and at very safe levels significantly reduced the growth of tumors in mice. Each antagonist and topotecan had additive effects to reduce cell viability with significant synergy demonstrated for the case of memantine. More significantly, combination treatments of xenografts in mice with ifenprodil and the chemotherapeutic agent topotecan produced clear additive effects that completely stopped tumor growth. Moreover, the ifenprodil and topotecan combination showed excellent supra-addition or synergy of inhibition for tumors ≤300 mm in size (P=4.7E−4). Combination treatment of memantine with topotecan also showed clear addition but, unlike ifenprodil, no synergy for the doses chosen. Conclusion Since topotecan is a drug of choice for treatment of rSCLC, our findings suggest that combining this agent with NMDA receptor blockade using the GluN2B antagonist, ifenprodil, will significantly improve patient outcomes., Video abstract

Background Small-cell lung cancer (SCLC) has a poor prognosis since there is currently no effective therapy for commonly recurring disease. In our previous study, both primary and recurrent human tumors have been shown to express functional N-methyl-D-aspartate (NMDA) receptors, and blockade of these receptors with GluN1 and GluN2B antagonists decreased tumor cell viability in vitro, and growth of tumor xenografts in nu/nu mice. Materials and methods In this study, we examine the influence of the GluN2B antagonist ifenprodil and the channel-blocker antagonist memantine, on cell viability and growth of tumor xenografts of recurrent SCLC (rSCLC) in mice. Results Both antagonists significantly reduced cell viability and levels of components of the ERK1/2 pathway, increased apoptosis, and at very safe levels significantly reduced the growth of tumors in mice. Each antagonist and topotecan had additive effects to reduce cell viability with significant synergy demonstrated for the case of memantine. More significantly, combination treatments of xenografts in mice with ifenprodil and the chemotherapeutic agent topotecan produced clear additive effects that completely stopped tumor growth. Moreover, the ifenprodil and topotecan combination showed excellent supra-addition or synergy of inhibition for tumors ≤300 mm in size (P=4.7E-4). Combination treatment of memantine with topotecan also showed clear addition but, unlike ifenprodil, no synergy for the doses chosen. Conclusion Since topotecan is a drug of choice for treatment of rSCLC, our findings suggest that combining this agent with NMDA receptor blockade using the GluN2B antagonist, ifenprodil, will significantly improve patient outcomes.

Cu-Based catalysts are currently used for the low-temperature water-gas shift (WGS) reaction, where the metallic Cu acts as the main active site. Under light irradiation, the Cu nanoparticles absorb photons, and electrons are promoted to the excited state. Then, the rapid relaxation of those excited electrons further drives the excited lattice phonon mode to generate heat in the local environment. This leads to a relatively high temperature at the catalytic reaction site. Herein, Cu-based catalysts were fabricated, and their catalytic activities for the WGS reaction were tested under the light irradiation of a xenon lamp without any additional heat treatment. Time-dependent temperature monitoring revealed that the temperature reached at 300 °C in 3 min, and then stabilized at ca. 350 °C for those Cu catalysts, which provided sufficient energy to overcome the energetic barrier and initiate the WGS reaction. Various metal oxides were optimized to function as the support, and Cu–ZnO exhibited the highest photothermal catalytic activity, which could produce 9.5 mmol of H2 in 30 min and had the CO conversion efficiency as high as 86.8%. The superior catalytic activity was ascribed to the strengthened interaction between the intermediate formate species and the partially oxidized Zn in the Cu–ZnO catalyst.

The richened reactive oxygen species (ROS) and their derived excessive inflammation at bone injured sites hinder osteogenesis of endosseous Ti-based implants. Herein, anti-oxidized polydopamine (PDA) is deposited on hydrothermal growth formed hydroxyapatite (HA) nanorods on Ti to form a core-shell structural nanorod-like array with HA as a core and PDA as an amorphous shell (PDA@HA), showing not only ROS scavenging ability but also near-infrared (NIR) light derived photo-thermal effects. PDA@HA suppresses inflammation based on its ROS scavenging ability to a certain extent, while periodic photo-thermal treatment (PTT) at a mild temperature (41 ± 1 °C) further accelerates the transition of the macrophages (MΦs) adhered to PDA@HA from the pro-inflammatory (M1) phenotype to the anti-inflammatory (M2) phenotype in vitro and in vivo. Transcriptomic analysis reveals that the activation of the PI3K-Akt1 signaling pathway is responsible for the periodic PTT induced acceleration of the M1-to-M2 transition of MΦs. Acting on mesenchymal stem cells (MSCs) with paracrine cytokines of M2 macrophages, PDA@HA with mild PTT greatly promote the osteogenetic functions of MSCs and thus osteogenesis. This work paves a way of employing mildly periodic PTT to induce a favorable immunomodulatory microenvironment for osteogenesis and provides insights into its underlying immunomodulation mechanism.

Power metering, relay protection, lightning current measurement, fault current measurement, and traveling current measurement in the field of power systems, as well as high-current measurement in the field of pulse power, require safe, high-accuracy, large-range, and wide-frequency-band current sensors. Current transformers and Rogowski coils can basically meet the requirements of measurement accuracy and range, but they are not good enough to meet the two requirements of safety and broad frequency bands. In this study, a non-contact broad-band current sensor is designed, which is based on the improvement of the traditional Rogowski coil to realize non-contact measurement of current. The equivalent circuit of the sensor was established, and the output characteristics of the sensor were calculated from the theoretical level as well as the design parameters were optimized. The sensor samples were made, and the usability of the sensor and the correctness of the theoretical calculation model were verified by 50 Hz AC current and 1.6 μs/3.4 μs impulse current tests. The non-contact current sensor designed in this paper has the advantages of simple structure, low cost, safety and reliability, large range, and wide operating band, and can be widely used for AC current or impulse current measurement in power systems and pulsed power fields.

Adult stem cells are critical for the maintenance of residential tissue homeostasis and functions. However, the roles of cellular protein homeostasis maintenance in stem cell proliferation and tissue homeostasis are not fully understood. Here, we find that Derlin-1 and TER94/VCP/p97, components of the endoplasmic reticulum (ER)-associated degradation (ERAD) pathway, restrain intestinal stem cell proliferation to maintain intestinal homeostasis in adult Drosophila. Depleting any of them results in increased stem cell proliferation and midgut homeostasis disruption. Derlin-1 is specifically localized in the ER of progenitors, and its C-terminus is required for its function. Interestingly, we find that increased stem cell proliferation is resulted from elevated ROS levels and activated JNK signaling in Derlin-1- or TER94-deficient progenitors. Further removal of reactive oxygen species (ROS) or inhibition of JNK signaling almost completely suppresses increased stem cell proliferation. Together, these data demonstrate that the ERAD pathway is critical for stem cell proliferation and tissue homeostasis. Thus, we provide insights into our understanding of the mechanisms underlying cellular protein homeostasis maintenance (ER protein quality control) in tissue homeostasis and tumor development.

BackgroundLidocaine administered through the working channel of a flexible bronchoscope can provide effective local anesthesia but cannot achieve good distribution in the airway. This study was undertaken to determine whether lidocaine delivered via a multi-orifice epidural catheter (three orifices/openings) is superior to conventional method and if a better distribution and decreased the cough reflex can be achieved.MethodsThe patients (N=100; 50 in each group) were randomized to receive either topical airway anesthesia by the “spray-as-you-go” technique via conventional application (group C) through the working channel of the bronchoscope or via a triple-orifice epidural catheter (group E). The primary outcome measurement was the cough severity, which was documented using a 4-point scale. Bronchoscopists and nurses assessed the coughing.ResultsThere was a significant difference in the median cough severity scores between the two groups (group C: 3 vs group E: 2, P =0.004). The median visual analogue scale (VAS) scores for the cough, were significantly higher in group C than those in group E (bronchoscopist: 3 vs 2 P =0.002; nurse: 3 vs 2, P

Carbapenem-resistant Klebsiella pneumoniae (CRKP), one of the major nosocomial pathogens, is increasingly becoming a serious threat to global public health. There is an urgent need to develop effective therapeutic and preventive approaches to combat the pathogen. Here, we identified and characterized a novel capsule depolymerase (K64-ORF41) derived from Klebsiella phage SH-KP152410, which showed specific activities for K. pneumoniae K64-serotype. We showed that this depolymerase could be used in the identification of K64 serotypes based on the capsular typing, and the results agreed well with those from the conventional serotyping method using antisera. From this study, we also identified K64 mutant strains, which showed typing discrepancy between wzi-sequencing based genotyping and depolymerase-based or antiserum-based typing methods. Further investigation indicated that the mutant strain has an insertion sequence (IS) in wcaJ, which led to the alteration of the capsular serotype structure. We further demonstrated that K64-ORF41 depolymerase could sensitize the bacteria to serum or neutrophil killing by degrading the capsular polysaccharide. In summary, the identified K64 depolymerase proves to be an accurate and reliable tool for capsular typing, which will facilitate the preventive intervention such as vaccine development. In addition, the polymerase may represent a potential and promising therapeutic biologics against CRKP-K64 infections.

Leptospirosis, caused by pathogenicLeptospiraspecies, has emerged as a widespread zoonotic disease worldwide. Macrophages mediate the elimination of pathogens through phagocytosis and cytokine production. Scavenger receptor A1 (SR-A1), one of the critical receptors mediating this process, plays a complicated role in innate immunity. However, the role of SR-A1 in the immune response against pathogenicLeptospirainvasion is unknown. In the present study, we found that SR-A1 is an important nonopsonic phagocytic receptor on murine macrophages forLeptospira. We also found that leptospiral LPS is the ligand of SR-A1. However, intraperitoneal injection of leptospires into WT mice presented with more severe jaundice, subcutaneous hemorrhaging, and higher bacteria burdens in blood and tissues than that of SR-A1-/-mice. Exacerbated cytokine and inflammatory mediator levels were also observed in WT mice and higher recruited macrophages in the liver than those of SR-A1-/-mice. Our findings collectively reveal that although beneficial in the uptake ofLeptospiraby macrophage, SR-A1 might be exploited byLeptospirato promote bacterial dissemination and modulate inflammatory activation, which causes a more severe infection in the host. These results provide our new insights into the innate immune response during early infection byL. interrogans.

Adult tissue homeostasis is maintained by residential stem cells. The proliferation and differentiation of adult stem cells must be tightly balanced to avoid excessive proliferation or premature differentiation. However, how stem cell proliferation is properly controlled remains elusive. Here, we find that auxilin (Aux) restricts intestinal stem cell (ISC) proliferation mainly through EGFR signaling. aux depletion leads to excessive ISC proliferation and midgut homeostasis disruption, which is unlikely caused by defective Notch signaling. Aux is expressed in multiple types of intestinal cells. Interestingly, aux depletion causes a dramatic increase in EGFR signaling, with a strong accumulation of EGFR at the plasma membrane and an increased expression of EGFR ligands in response to tissue stress. Furthermore, Aux co-localizes and associates with EGFR. Finally, blocking EGFR signaling completely suppresses the defects caused by aux depletion. Together, these data demonstrate that Aux mainly safeguards EGFR activation to keep a proper ISC proliferation rate to maintain midgut homeostasis.

Mestrado em Marketing Com o desenvolvimento da ciência e da tecnologia na sociedade moderna, cada vez mais pessoas estão a usar smartphones. Ao mesmo tempo, nos últimos anos, têm aparecido várias aplicações de entrega de comida ao domicílio em Portugal e na China. Por exemplo, o Glovo e o Ubereats são as aplicações mais usadas em Portugal, e o Meituan e o Eleme as mais usadas na China. Este trabalho tem como objetivo analisar as perceções e fatores influenciadores na compra de serviços de entrega de comida ao domicílio pelos consumidores em Portugal e na China, focando-se principais diferenças entre os consumidores dos dois países. Para esta investigação recorreu-se a um estudo quantitativo, através da aplicação de um questionário online para recolher dados, que foi dividido em duas versões, uma em português e outra em chinês. A técnica de amostra utilizada foi não probabilística por conveniência, com a criação do efeito bola de neve. Finalmente obteve-se um total de 293 respostas válidas. Entre estas, as respostas efetivas dos consumidores chineses e dos consumidores portugueses foram 206 e 87, respetivamente. Os resultados obtidos indicam que para os consumidores portugueses, a perceção da confidencialidade de informação pessoal está positivamente relacionada com a confiança nos serviços de entrega de comida ao domicílio. Para os consumidores chineses, exceto a perceção da segurança durante a entrega, todos outros fatores, como a perceção da segurança alimentar, a perceção da economia de tempo, a perceção da política promocional, e perceção da conveniência e a perceção da confidencialidade de informação pessoal, estão positivamente associados com a mesma. Adicionalmente, a confiança estápositivamente relacionada com a intenção de compra deste serviço tanto pelos consumidores portugueses como chineses. Como o serviço de entrega de comida ao domicílio é uma indústria emergente, existe até ao momento poucas estudos e investigações na área. Assim sendo esta dissertação é uma contribuição para literatura existente a nível académico, e ao mesmo tempo, um registo da atual indústria para estas plataformas em Portugal e na China. With the development of science and technology in our modern society, more and more people are using smartphones. At the same time, in recent years, there have been developed several mobile applications for home delivery food both in Portugal and China. For example, Glovo and Ubereats are the most used applications in Portugal, and Meituan and Eleme the most used in China. This work aims to analyze the perceptions and influencing factors for consumers in Portugal and China during the purchase of food delivery services, focusing on the differences between consumers in both countries. For this investigation a quantitative study was used, with the application of an online questionnaire to collect data, which was divided into two versions, one in Portuguese and the other in Chinese. The sample technique used was non-probabilistic for convenience, with the creation of the snowball effect. Finally, a total of 293 valid responses were obtained. Among these, the effective responses of Chinese consumers and those of Portuguese consumers are 206 and 87, respectively. The results obtained indicate that for Portuguese consumers, the perception of confidentiality of personal information is positively related to trust in home delivery services. For Chinese consumers, except for the perception of security during delivery, all other factors, such as the perception of food safety, the perception of time savings, the perception of promotional policy, and the perception of convenience and the perception of confidentiality of personal information , are positively associated with it. Additionally, trust is positively related to the intention of purchase of such service by Portuguese and Chinese consumers. Seen as home delivery service is an emerging industry, there is so far little research and experience in this area. Therefore, this study is also a contribution to the existing academic literature, and at the same time, a report of the current industry for these platforms both in Portugal and China. info:eu-repo/semantics/publishedVersion

Ganoderic Acid A (GA) has many pharmacological effects such as anti-tumor, antibacterial, anti-inflammatory, and immunosuppressive effects. However, the protective effect of GA on liver injury has not been reported. This study aimed to investigate the action of GA on insufficient methionine and choline combined with high-fat diet (HFD)-induced non-alcoholic fatty liver disease (NAFLD) in rats. NAFLD model was established by insufficient methionine and choline combined with high fat feeding to rats. The levels of Acetyl-CoA carboxylase, fatty acid synthase, sterol regulatory element binding protein, liver X receptors, AMP-activated protein kinase, peroxisome proliferator-activated receptor α, PPARg coactivator 1α and NF-κB pathway in the liver were detected by western blot. The results of this study demonstrated that the expression of GA can not only significantly decrease the live weight and liver weight per body weight of HFD mice, but also restore the alanine aminotransferase, aspartate aminotransferase, total bilirubin levels, triglyceride and cholesterol in serum. In addition, the expression of GA increased the levels of high-density lipoprotein cholesterol in serum, ameliorated pathological changes and decreased NAS score of mice's liver. In conclusion, the treatment with GA could improve NAFLD in rats by regulating the levels of signaling events involved in free fatty acid production, lipid oxidation and liver inflammation.

Bangia fuscopurpurea is a farmed species in the ancient family Bangiales. It inhabits upper intertidal zones and suffers periodical desiccation and osmotic stress. The transcriptomic regulation under dehydration and hyposalinity was investigated. The differentially expressed genes (DEGs) accounted for 18.7% of the unigenes obtained by de novo transcriptome assembly (|log2fold-change| ≥ 1, FDR ≤ 0.001). Over 72% of the DEGs were downregulated under stress. The DEGs were predominantly enriched into the KEGG pathways “metabolic pathways,” “ribosome,” “biosynthesis of secondary metabolites,” “protein processing in endoplasmic reticulum,” and “oxidative phosphorylation.” The optimum photosynthetic efficiency (Fv/Fm) and photochemical quenching (qP) dropped significantly with 89% relative water loss and recovered rapidly after being rehydrated. Most DEGs regarding “photosynthesis” and “C3 carbon fixation” were upregulated in the dehydrated thalli, which may enable the thalli to gain photosynthetic recovery once being rehydrated. Fv/Fm and qP decreased significantly with 1 h of 90% freshwater treatment and then recovered to the control level 1 day later. With 6 h hyposaline treatment, expression of plasma membrane H+-ATPase genes was strongly and predominantly induced while the mRNA abundance of vacuolar, chloroplastic, and mitochondrial H+-ATPase genes decreased or showed no significant change. Some transporter, ion channel, and transmembrane protein genes together with the gene-encoding key enzymes involving in proline and heteroside metabolism were upregulated under hyposalinity. The results indicated that transmembrane exchange of ion and osmolytes was induced under hyposalinity to balance the osmotic fluctuation, which seemed to be triggered by plasma membrane H+-ATPases. These findings will facilitate elucidating the stress acclimation mechanism of B. fuscopurpurea.

Continuous and intensive selective breeding reduces the genetic diversity of cultivated Pyropia stock, thereby increasing its susceptibility to diseases, and hindering the sustainable development of the Pyropia industry. To develop new Pyropia germplasms with desirable agronomic traits, a new Pyropia yezoensis strain, “Huangyou No. 1” was bred from a wild subtidal population. In this study, the genetic diversity of this strain was analyzed using AFLP. Totally, 249 loci were obtained, of which 248 were polymorphic with a polymorphism rate of 99.6%, using seven primer pairs. The intra-population polymorphism rates of the populations Wild 1 and Wild 2 were the highest (47.39% and 59.44%, respectively), while they were relatively low (20.88–24.5%) in the cultivated populations. The genetic distances between the breeding Pyropia populations cultivated in the same aquaculture farm were low. Specifically, the F2 offspring of “Huangyou No. 1” and the control cultivar, cultivated using the “never-drying” protocol, were genetically distant from the other populations, including that from the same seedlings, yet cultivated using the “periodic-drying” protocol. The never-drying cultivation protocol had a high likelihood of influencing genetic diversity of Pyropia yezoensis.