Application of a derivative of human defensin 5 to treat ionizing radiation-induced enterogenic infection.

Enterogenic infection is a common complication for patients with radiation injury and requires efficient therapeutics in the clinic. Herein, we evaluated the promising drug candidate T7E21RHD5, which is a peptide derived from intestinal Paneth cell-secreted human defensin 5. Oral administration of this peptide alleviated the diarrhea symptoms of mice that received total abdominal irradiation (TAI, γ-ray, 12 Gy) and improved survival. Pathologic analysis revealed that T7E21RHD5 elicited an obvious mitigation of ionizing radiation (IR)-induced epithelial damage and ameliorated the reduction in the levels of claudin, zonula occluden 1 and occludin, three tight junction proteins in the ileum. Additionally, T7E21RHD5 regulated the gut microbiota in TAI mice by remodeling ß diversity, manifested as a reversal of the inverted proportion of Bacteroidota to Firmicutes caused by IR. T7E21RHD5 treatment also decreased the abundance of pathogenic Escherichia-Shigella but significantly increased the levels of Alloprevotella and Prevotellaceae_NK3B31, two short-chain fatty acid-producing bacterial genera in the gut. Accordingly, the translocation of enterobacteria and lipopolysaccharide to the blood, as well as the infectious inflammatory responses in the intestine after TAI, was all suppressed by T7E21RHD5 administration. Hence, this versatile antimicrobial peptide possesses promising application prospects in the treatment of IR-induced enterogenic infection.

Cantilever beam bending as a potential method to determine the elasticity of cooked Udon noodles.

Elasticity is a critical measure of the eating quality of Udon noodles. To characterize the elasticity of Udon noodles, an instrumental method based on the cantilever beam bending test was established. Firstly, the optimum test parameters were determined. Then, texture profile analysis, compression, tension, and cantilever beam bending methods were used to measure the elasticity of 25 commercial Udon noodles with different shapes and sizes, and the correlations between elasticity obtained by the above instrumental methods and sensory evaluation were analyzed. Finally, how the shape and size of Udon noodles influenced their elasticity was discussed in detail. Within the deflection of 2.0 mm, the force increased approximately linearly with increasing deflection, and moderate loading speed (0.5-1.0 mm/s) should be used in the cantilever beam bending experiments to improve the accuracy of results. The bending stiffness obtained by the cantilever beam bending method exhibited a higher coefficient of variation and stronger correlation with the elasticity of sensory evaluation than other instrumental methods. Furthermore, the Udon noodle sample with a higher size, especially the thickness, had higher elasticity, and the Udon noodle sample with a rectangular cross-section showed higher elasticity than that with a circular cross-section. In conclusion, the bending stiffness determined by the cantilever beam bending method could be used to characterize the elasticity of cooked Udon noodles.

Coral Detection, Ranging, and Assessment (CDRA) algorithm-based automatic estimation of coral reef coverage.

Climate change poses significant threats to the health of coral reefs. The detection of coral coverage is a quantitative method for assessing the health of coral reefs. Valuable insights into the effects of climate change on coral reef ecosystems and the trends in coral reef development can be gained through the monitoring of coral coverage. However, traditional manual methods for estimating coral coverage are time-consuming and labor-intensive, limiting the scalability and efficiency of coral surveys. The field of automated processing for coral video imagery is currently limited, impeding the development of effective techniques for comprehensive coral monitoring and further hindering progress in coral ecosystem monitoring. To cope with these challenges, there is a crucial need for automated systems to efficiently handle large volumes of video data, thereby transforming imaging devices, such as underwater robots, into autonomous sensors capable of conducting reliable and comprehensive surveys. In response to this pressing issue, this study presents a novel video analysis approach, termed the Coral Detection, Ranging, and Assessment (CDRA) algorithm, which combines cutting-edge computer vision techniques including You Only Look Once version 5 (YOLOv5) object detection, binocular stereo vision, and tracking algorithms. The CDRA algorithm aims to analyze continuous video segments of coral reefs, enabling the identification, counting, and estimation of size and location of individual coral colonies. Experimental evaluations conducted in controlled pool environments demonstrated the high accuracy of the proposed method, with an average accuracy of 90.7% in estimating coral coverage at different locations. Moreover, field trials conducted in Xidao Island of China verified the effectiveness and robustness of the CDRA algorithm under varying water quality and lighting conditions. The findings of this study represent a significant step towards the development of reliable and automated techniques for coral reef detection, contributing to the advancement of coral reef conservation efforts.

Harnessing PUF-Based Reporters for Noninvasive Imaging of the MicroRNA Dynamics in Differentiation.

Precise characterization of miRNA expression patterns is critical to exploit the complexity of miRNA regulation in biology. Herein, we developed a Pumilio/FBF (PUF) protein-based engineering luciferase reporter system, PUF/miR, to quantitatively and non-invasively sense miRNA activity in living cells and animal models. We verified the feasibility of this reporter by monitoring the expression of several types of miRNAs (miRNA-9, 124a, 1, and 133a) in neural and muscle differentiated cells as well as subcutaneous or tibial anterior muscles in mice. The quantitative RT-PCR also validated the reliability and quantitative consistency of bioluminescence imaging in detecting miRNA expression. We further effectively employed this reporter system to visualize the expression of miRNA-1 and miRNA-133a in mouse models of skeletal muscle injury. As a non-invasive and convenient innovative approach, our results have realized the positive bioluminescence imaging of endogenous miRNAs in vitro and in vivo using the PUF/miR system. We believe that this approach would provide a potential means for noninvasive monitoring of disease-related miRNAs and could facilitate a deeper understanding of miRNA biology.

Analysis of the effect of rolling speed on the texture properties of noodle dough from water-solid interaction, development of gluten network, and bubble distribution.

To explore the influence of rolling speed on texture property of the noodle dough with 34% and 38% water additions, the water status, development of gluten network, and bubble distribution were studied. The dough sheets with 38% water addition had more uniform gluten network, and fewer bubbles, but lower hardness and tensile force because of the softening effect of water. As rolling speed increased, the water mobility increased, gluten network became weakened, and bubble number increased, leading to decreases in texture properties. Dough sheets with 38% water addition were less responsive to changes in rolling speed than that with 34% water addition because of the higher hydration of gluten. With increasing rolling speed, the total length and number of gluten network lines in length direction (rolling direction) reduced more than those in width direction, corresponding to the greater reductions in the texture properties of the dough sheet in length direction.

Irradiation accelerates SARS-CoV-2 infection by enhancing sphingolipid metabolism.

Cancer patients who receive radiotherapy have a high risk of severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) infection, but the concrete reason remains unclear. Herein, we investigated the influence of irradiation on the vulnerability of cancer cells to SARS-CoV-2 using S pseudovirions and probed the underlying mechanism via RNA-seq and other molecular biology techniques. Owing to the enhancement of sphingolipid metabolism, irradiation accelerated pseudovirion infection. Mechanistically, irradiation induced the expression of acid sphingomyelinase (ASM), which catalyses the hydrolysis of sphingomyelin to ceramide, contributing to lipid raft formation and promoting SARS-CoV-2 invasion. Inhibition of lipid raft formation with methyl-ß-cyclodextrin (MßCD) or the tyrosine kinase inhibitor genistein and ASM suppression through small interfering RNA or amitriptyline (AMT) treatment abolished the enhancing effect of irradiation on viral infection. Animal experiments supported the finding that irradiation promoted SARS-CoV-2 S pseudovirion infection in A549 cell tumour-bearing BALB/c nude mice, whereas AMT treatment dramatically decreased viral infection. This study discloses the role of sphingolipid metabolism in irradiation-induced SARS-CoV-2 infection, thus providing a potential target for clinical intervention to protect patients receiving radiotherapy from COVID-19.

Caenorhabditis elegans as an emerging model in food and nutrition research: importance of standardizing base diet.

As a model organism that has helped revolutionize life sciences, Caenorhabditis elegans has been increasingly used in nutrition research. Here we explore the tradeoffs between pros and cons of its use as a dietary model based primarily on literature review from the past decade. We first provide an overview of its experimental strengths as an animal model, focusing on lifespan and healthspan, behavioral and physiological phenotypes, and conservation of key nutritional pathways. We then summarize recent advances of its use in nutritional studies, e.g. food preference and feeding behavior, sugar status and metabolic reprogramming, lifetime and transgenerational nutrition tracking, and diet-microbiota-host interactions, highlighting cutting-edge technologies originated from or developed in C. elegans. We further review current challenges of using C. elegans as a nutritional model, followed by in-depth discussions on potential solutions. In particular, growth scales and throughputs, food uptake mode, and axenic culture of C. elegans are appraised in the context of food research. We also provide perspectives for future development of chemically defined nematode food ("NemaFood") for C. elegans, which is now widely accepted as a versatile and affordable in vivo model and has begun to show transformative potential to pioneer nutrition science.

In Vivo Visualization of RNAi Efficiency Using a Pumilio/FBF Protein-Based Reporter.

As a strategy that induces gene silencing by the delivery of small interfering RNA (siRNA) targeting a specific gene locus into cells or tissues, RNA interference (RNAi) technology holds the potential to be a powerful tool in a range of intractable disorder therapeutics. However, reliable noninvasive probes for visualizing the siRNA delivery and silencing efficiency have become a major obstacle in siRNA-based treatment. Here, we describe the development of an RNA-binding protein Pumilio/FBF (PUF)-based reporter probe for the monitoring of siRNA delivery efficiency and functional screening of effective siRNA target sites in vivo. This reporter consisted of a Firefly luciferase (Fluc) gene whose expression is regulated by the unique interaction architecture of the PUF protein with its Nanos response element (NRE) target RNA. We showed that a robust and rapid increase in the luminescence signal was detected by the successful delivery of siRNA against the enhanced green fluorescent protein (EGFP) or p53 genes into mammalian cells or the livers of mice. The delivery efficiencies of various commercial transfection vehicles were quantitatively evaluated with this reporter. In addition, we also employed in vivo bioluminescence imaging to screen and identify the most potent siRNA targeting p53. Our study indicates that the positive-readout reporter represents a promising indicator for siRNA optimization and visualization, advancing the development of siRNA therapeutic products.

Assessment of the influence of gluten quality on highland barley dough sheet quality by different instruments.

This study was to compare the results of texture analyzer with those of farinograph and extensograph and determine whether texture analyzer could be used to evaluate the processing quality of highland barley flour (HBF) dough sheet. The farinograph and extensograph tests were used to determine the reconstituted flour properties, a texture analyzer was applied to measure the tensile strength (TS) of HBF dough sheet, and the content of glutenin macropolymer (GMP), free sulfhydryl (-SH) and secondary structure of protein and microstructure in HBF dough sheet were investigated. Furthermore, correlations between these parameters were determined by regression analysis and Pearson correlation coefficient. It was suggested that the reconstituted flours with a higher gluten index showed a higher farinograph quality number (FQN) and greater maximum resistance to extension (Rm ). HBF dough sheets with higher gluten index possessed higher GMP and lower free -SH contents, a more ordered secondary structure of protein, resulting in a more compact gluten network and a stronger TS. The regression and correlation analysis showed that TS was positively correlated with FQN and Rm . In addition, it was significantly correlated with the content of GMP, -SH, secondary structure of protein and gluten network. It was concluded that texture analyzer could be an alternative approach to evaluate the processing quality of HBF dough sheet. Moreover, the gluten index of flours could be used to predict the processing quality of HBF dough sheet.

Caenorhabditis elegans as a Model System for Discovering Bioactive Compounds Against Polyglutamine-Mediated Neurotoxicity.

Age-related misfolding and aggregation of pathogenic proteins are responsible for several neurodegenerative diseases. For example, Huntington's disease (HD) is principally driven by a CAG nucleotide repeat that encodes an expanded glutamine tract in huntingtin protein. Thus, the inhibition of polyglutamine (polyQ) aggregation and, in particular, aggregation-associated neurotoxicity is a useful strategy for the prevention of HD and other polyQ-associated conditions. This paper introduces generalized experimental protocols to assess the neuroprotective capacity of test compounds against HD using established polyQ transgenic Caenorhabditis elegans models. The AM141 strain is chosen for the polyQ aggregation assay as an age-associated phenotype of discrete fluorescent aggregates can be easily observed in its body wall at the adult stage due to muscle-specific expression of polyQ::YFP fusion proteins. In contrast, the HA759 model with strong expression of polyQ-expanded tracts in ASH neurons is used to examine neuronal death and chemoavoidance behavior. To comprehensively evaluate the neuroprotective capacity of target compounds, the above test results are ultimately presented as a radar chart with profiling of multiple phenotypes in a manner of direct comparison and direct viewing.

The immediate and subsequent effects of public health interventions for COVID-19 on the leisure and recreation industry.

Public health interventions to combat COVID-19 can be viewed as an exogenous shock to the economy, especially for industries-such as leisure, recreation, and tourism-that rely heavily on human mobility. This study investigates whether and how exactly the economic impact of government public health policies varies over time. Focusing on the leisure and recreation industry, we use data for 131 countries/regions from February to May 2020 and employ generalized difference-in-differences models to investigate the short- and longer-term effects of public health policies. We find that stricter policies lead, on average, to an immediate 9.2-percentage-point drop in leisure and recreation participation. Even so, that industry recovers in about seven weeks after a COVID-19 outbreak in countries/regions that undertake active interventions. After thirteen weeks, leisure and recreation involvement recovers to 70% of pre-pandemic levels in a place that actively intervened but stagnates at about 40% in one that did not.

The impact of starch degradation induced by pre-gelatinization treatment on the quality of noodles.

A study was to determine the correlation between the fine structure and the physicochemical properties of pre-gelatinized corn starch (PGCS) and the quality of noodles with added PGCS. Drum-dried corn starch (DDCS) and extrusion-cooked corn starch (ECCS) were used. The results revealed that the cold-water viscosity of ECCS and DDCS was about 10 and 100 times greater, respectively, than that of native corn starch (NCS), and the gel strength of DDCS was 1.67 times greater than that of ECCS. The average hydrodynamic radius of whole molecular (R¯h) and Ap were approximately half that of native corn starch in ECCS. Dough sheets prepared with various concentrations of PGCS (5-10%, w/w) had significantly greater tensile strength, and the addition of PGCS significantly increased the smoothness of the cooked noodles. DDCS was better than ECCS for increasing the noodles' processing quality.

Softness, elasticity, and smoothness characteristics of cooked udon noodles based on texture analysis.

Objective evaluation methods for the elasticity, smoothness, and softness of cooked udon noodles were established on the basis of texture analysis. Noodles with different diameters and amylose contents were prepared to verify the reliability of the proposed evaluation methods. Results revealed that the elasticity of udon noodles could be examined by using the method that involved rinsing cooked noodles with 10°C cold water for 30 s and stretching a single strand of cooked noodle at 3 mm/s with an A/KIE probe until broken. The stiffness of the sample at 3.5 mm could be determined to indicate the elasticity caused by the size of dried udon noodles. The softness and smoothness of cooked udon noodles could be examined when cooled and drained for 30 s by using an heavy duty platform/pasta firmness/stickiness rig probe. The elasticity of cooked udon noodles increased as their diameter increased, and their smoothness and softness significantly increased as the amylose content decreased (p < .05). Texture analysis could be used to quickly and accurately indicate the elasticity, softness, and smoothness of cooked udon noodles. The texture characteristics of udon noodles were related to their formulation, processing, and size. Establishing an adaptability evaluation method for the quality of udon noodles is a prerequisite for optimizing processing technologies and developing new products. Thus far, limited research has focused on objective methods for evaluating the texture of cooked udon noodles. This work developed valuable instrumental methods for examining the elasticity, softness, and smoothness of cooked udon noodles and provided noodle manufacturers a tool for selecting udon noodle formulations and processing technology based on texture analysis.

Implementation of the CRISPR-Cas13a system in fission yeast and its repurposing for precise RNA editing.

In contrast to genome editing, which introduces genetic changes at the DNA level, disrupting or editing gene transcripts provides a distinct approach to perturbing a genetic system, offering benefits complementary to classic genetic approaches. To develop a new toolset for manipulating RNA, we first implemented a member of the type VI CRISPR systems, Cas13a from Leptotrichia shahii (LshCas13a), in Schizosaccharomyces pombe, an important model organism employed by biologists to study key cellular mechanisms conserved from yeast to humans. This approach was shown to knock down targeted endogenous gene transcripts with different efficiencies. Second, we engineered an RNA editing system by tethering an inactive form of LshCas13a (dCas13) to the catalytic domain of human adenosine deaminase acting on RNA type 2 (hADAR2d), which was shown to be programmable with crRNA to target messenger RNAs and precisely edit specific nucleotide residues. We optimized system parameters using a dual-fluorescence reporter and demonstrated the utility of the system in editing randomly selected endogenous gene transcripts. We further used it to restore the transposition of retrotransposon Tf1 mutants in fission yeast, providing a potential novel toolset for retrovirus manipulation and interference.

Effect of egg freshness on their automatic orientation.

BACKGROUND: High-quality eggs in unified packaging are desired by egg production enterprises. Automatic orientation apparatus is frequently used to orient eggs uniformly to pointed-end-down position for packaging. However, such apparatus may not work accordingly if the eggs are stored under improper methods or for excessive storage time. To study the effect of egg freshness on the efficiency of automatic orientation process, the relationship between egg freshness and its orientation motions was investigated under different storage conditions. RESULTS: The results showed that as the storage time increased, centroid position and pointed-end-down turnover ratio decreased; other parameters such as eggs' obliquity at stationary state, horizontal deflection angle, speed, acceleration of axial motion, side-slip angle and rolling distance increased. However, the effects of storage time on the guiding distance of the guide rod were not apparent. In addition, the higher the storage temperature, the greater the changes of the final orientation states of eggs on the conveyor line. If the eggs were to be oriented uniformly, they should be stored for less than 25, 16, 10 and 7 days at 10 °C, 18 °C, 26 °C and 34 °C, respectively, under a relative humidity of 75%. CONCLUSION: The experimental results presented in this paper are very useful for quality control and quality assurance in egg production enterprises. © 2017 Society of Chemical Industry.

Lipoxin A4 and platelet activating factor are involved in E. coli or LPS-induced lung inflammation in CFTR-deficient mice.

CFTR (cystic fibrosis transmembrane conductance regulator) is expressed by both neutrophils and platelets. Lack of functional CFTR could lead to severe lung infection and inflammation. Here, we found that mutation of CFTR (F508del) or inhibition of CFTR in mice led to more severe thrombocytopenia, alveolar neutrocytosis and bacteriosis, and lower lipoxin A4/MIP-2 (macrophage inhibitory protein-2) or lipoxin A4/neutrophil ratios in the BAL (bronchoalveolar lavage) during acute E. coli pneumonia. In vitro, inhibition of CFTR promotes MIP-2 production in LPS-stimulated neutrophils; however, lipoxin A4 could dose-dependently suppress this effect. In LPS-induced acute lung inflammation, blockade of PSGL-1 (P-selectin glycoprotein ligand-1) or P-selectin, antagonism of PAF by WEB2086, or correction of mutated CFTR trafficking by KM11060 could significantly increase plasma lipoxin A4 levels in F508del relevant to wildtype mice. Concurrently, F508del mice had higher plasma platelet activating factor (PAF) levels and PAF-AH activity compared to wildtype under LPS challenge. Inhibiting hydrolysis of PAF by a specific PAF-AH (PAF-acetylhydrolase) inhibitor, MAFP, could worsen LPS-induced lung inflammation in F508del mice compared to vehicle treated F508del group. Particularly, depletion of platelets in F508del mice could significantly decrease plasma lipoxin A4 and PAF-AH activity and deteriorate LPS-induced lung inflammation compared to control F508del mice. Taken together, lipoxin A4 and PAF are involved in E. coli or LPS-induced lung inflammation in CFTR-deficient mice, suggesting that lipoxin A4 and PAF might be therapeutic targets for ameliorating CFTR-deficiency deteriorated lung inflammation.

Important role of platelets in modulating endotoxin-induced lung inflammation in CFTR-deficient mice.

Mutation of CFTR (cystic fibrosis transmembrane conductance regulator) leads to cystic fibrosis (CF). Patients with CF develop abnormalities of blood platelets and recurrent lung inflammation. However, whether CFTR-mutated platelets play a role in the development of lung inflammation is elusive. Therefore, we intratracheally challenged wildtype and F508del (a common type of CFTR mutation) mice with LPS to observe changes of F508del platelets in the peripheral blood and indexes of lung inflammation (BAL neutrophils and protein levels). Furthermore, we investigated whether or not and how F508del platelets modulate the LPS-induced acute lung inflammation by targeting anti-platelet aggregation, depletion of neutrophils, reconstitution of bone marrow or neutrophils, blockade of P-selectin glycoprotein ligand-1 (PSGL-1), platelet activating factor (PAF), and correction of mutated CFTR trafficking. We found that LPS-challenged F508del mice developed severe thrombocytopenia and had higher levels of plasma TXB2 coincided with neutrophilic lung inflammation relative to wildtype control. Inhibition of F508del platelet aggregation or depletion of F508del neutrophils diminished the LPS-induced lung inflammation in the F508del mice. Moreover, wildtype mice reconstituted with either F508del bone marrow or neutrophils developed worse thrombocytopenia. Blocking PSGL-1, platelet activating factor (PAF), or rectifying trafficking of mutated CFTR in F508del mice diminished and alveolar neutrophil transmigration in the LPS-challenged F508del mice. These findings suggest that F508del platelets and their interaction with neutrophils are requisite for the development of LPS-induced lung inflammation and injury. As such, targeting platelets might be an emerging strategy for dampening recurrent lung inflammation in cystic fibrosis patients.