Influenza A virus inhibits TET2 expression by endoribonuclease PA-X to attenuate type I interferon signaling and promote viral replication.

Influenza A virus (IAV) expresses several accessory proteins to limit host anti-viral restriction factors to facilitate viral replication. The Ten-Eleven Translocation 2 (TET2) is a methylcytosine dioxygenase that promotes DNA demethylation by catalyzing the oxidation of 5-methylcytosine (5mC) into 5-hydroxymethylcytosine (5hmC), which plays a vital role in hematopoiesis and immunity. Here we report that TET2 is a host restriction factor that limits IAV replication. But IAV endoribonuclease PA-X is able to remove the replication restriction by binding to TET2 mRNA and driving TET2 mRNA degradation to reduce TET2 expression during infection. Genetic inactivation of TET2 markedly enhances IAV replication in vitro and in vivo. Mechanistically, we found that TET2 regulates demethylation and transcription of STAT1 and some interferon-stimulated genes (ISGs), including ISG15, ISG20, and IFIT5, so the loss of TET2 greatly impairs type I Interferon signaling. Furthermore, we confirmed that TET2-mediated demethylation of the STAT1 gene is critical for interferon anti-viral activity. Our study demonstrates that the host TET2 is essential to the innate immune response against IAV infection.

Recent Advances in Pristine Iron Triad Metal-Organic Framework Cathodes for Alkali Metal-Ion Batteries.

Pristine iron triad metal-organic frameworks (MOFs), i.e., Fe-MOFs, Co-MOFs, Ni-MOFs, and heterometallic iron triad MOFs, are utilized as versatile and promising cathodes for alkali metal-ion batteries, owing to their distinctive structure characteristics, including modifiable and designable composition, multi-electron redox-active sites, exceptional porosity, and stable construction facilitating rapid ion diffusion. Notably, pristine iron triad MOFs cathodes have recently achieved significant milestones in electrochemical energy storage due to their exceptional electrochemical properties. Here, the recent advances in pristine iron triad MOFs cathodes for alkali metal-ion batteries are summarized. The redox reaction mechanisms and essential strategies to boost the electrochemical behaviors in associated electrochemical energy storage devices are also explored. Furthermore, insights into the future prospects related to pristine iron triad MOFs cathodes for lithium-ion, sodium-ion, and potassium-ion batteries are also delivered.

Recommendation of a standardized broth microdilution method for antimicrobial susceptibility testing of Avibacterium paragallinarum and resistance monitoring.

This study aimed to develop a method for standardized broth microdilution antimicrobial susceptibility testing (AST) of Avibacterium (Av.) paragallinarum, the causative agent of infectious coryza in chickens. For this, a total of 83 Av. paragallinarum isolates and strains were collected from 15 countries. To select unrelated isolates for method validation steps, macrorestriction analyses were performed with 15 Av. paragallinarum. The visible growth of Av. paragallinarum was examined in six broth media and growth curves were compiled. In Veterinary Fastidious Medium and cation-adjusted Mueller-Hinton broth (CAMHB) + 1% chicken serum + 0.0025% NADH (CAMHB + CS + NADH), visible growth of all isolates was detected and both media allowed adequate bacterial growth. Due to the better readability of Av. paragallinarum growth in microtiter plates, CAMHB + CS + NADH was chosen for AST. Repetitions of MIC testing with five epidemiologically unrelated isolates using a panel of 24 antimicrobial agents resulted in high essential MIC agreements of 96%-100% after 48-h incubation at 35 ± 2°C. Hence, the remaining 78 Av. paragallinarum were tested and demonstrated easily readable MICs with the proposed method. Differences in MICs were detected between isolates from different continents, with isolates from Africa showing lower MICs compared to isolates from America and Europe, which more often showed elevated MICs of aminoglycosides, quinolones, tetracyclines, and/or trimethoprim/sulfamethoxazole. PCR analyses of isolates used for method development revealed that isolates with elevated MICs of tetracyclines harbored the tetracycline resistance gene tet(B) but none of the other tested resistance genes were detected. Therefore, whole-genome sequencing data from 62 Av. paragallinarum were analyzed and revealed the presence of sequences showing nucleotide sequence identity to the genes aph(6)-Id, aph(3″)-Ib, blaTEM-1B, catA2, sul2, tet(B), tet(H), and mcr-like. Overall, the proposed method using CAMHB + CS + NADH for susceptibility testing with 48-h incubation time at 35 ± 2°C in ambient air was shown to be suitable for Av. paragallinarum. Due to a variety of resistance genes detected, the development of clinical breakpoints is highly recommended. IMPORTANCE: Avibacterium paragallinarum is an important pathogen in veterinary medicine that causes infectious coryza in chickens. Since antibiotics are often used for treatment and resistance of the pathogen is known, targeted therapy should be given after resistance testing of the pathogen. Unfortunately, there is currently no accepted method in standards that allows susceptibility testing of this fastidious pathogen. Therefore, we have worked out a method that allows harmonized susceptibility testing of the pathogen. The method meets the requirements of the CLSI and could be used by diagnostic laboratories.

A Channel-Dimensional Feature-Reconstructed Deep Learning Model for Predicting Breast Cancer Molecular Subtypes on Overall b-Value Diffusion-Weighted MRI.

BACKGROUND: Dynamic contrast-enhanced (DCE) MRI commonly outperforms diffusion-weighted (DW) MRI in breast cancer discrimination. However, the side effects of contrast agents limit the use of DCE-MRI, particularly in patients with chronic kidney disease. PURPOSE: To develop a novel deep learning model to fully exploit the potential of overall b-value DW-MRI without the need for a contrast agent in predicting breast cancer molecular subtypes and to evaluate its performance in comparison with DCE-MRI. STUDY TYPE: Prospective. SUBJECTS: 486 female breast cancer patients (training/validation/test: 64%/16%/20%). FIELD STRENGTH/SEQUENCE: 3.0 T/DW-MRI (13 b-values) and DCE-MRI (one precontrast and five postcontrast phases). ASSESSMENT: The breast cancers were divided into four subtypes: luminal A, luminal B, HER2+, and triple negative. A channel-dimensional feature-reconstructed (CDFR) deep neural network (DNN) was proposed to predict these subtypes using pathological diagnosis as the reference standard. Additionally, a non-CDFR DNN (NCDFR-DNN) was built for comparative purposes. A mixture ensemble DNN (ME-DNN) integrating two CDFR-DNNs was constructed to identify subtypes on multiparametric MRI (MP-MRI) combing DW-MRI and DCE-MRI. STATISTICAL TESTS: Model performance was evaluated using accuracy, sensitivity, specificity, and area under the receiver operating characteristic curve (AUC). Model comparisons were performed using the one-way analysis of variance with least significant difference post hoc test and the DeLong test. P < 0.05 was considered significant. RESULTS: The CDFR-DNN (accuracies, 0.79 ~ 0.80; AUCs, 0.93 ~ 0.94) demonstrated significantly improved predictive performance than the NCDFR-DNN (accuracies, 0.76 ~ 0.78; AUCs, 0.92 ~ 0.93) on DW-MRI. Utilizing the CDFR-DNN, DW-MRI attained the predictive performance equal (P = 0.065 ~ 1.000) to DCE-MRI (accuracies, 0.79 ~ 0.80; AUCs, 0.93 ~ 0.95). The predictive performance of the ME-DNN on MP-MRI (accuracies, 0.85 ~ 0.87; AUCs, 0.96 ~ 0.97) was superior to those of both the CDFR-DNN and NCDFR-DNN on either DW-MRI or DCE-MRI. DATA CONCLUSION: The CDFR-DNN enabled overall b-value DW-MRI to achieve the predictive performance comparable to DCE-MRI. MP-MRI outperformed DW-MRI and DCE-MRI in subtype prediction. LEVEL OF EVIDENCE: 2 TECHNICAL EFFICACY STAGE: 1.

Multidentate Molecule Anchoring Halide Perovskite Surface and Regulating Crystallization Kinetics toward Efficient Light-Emitting Diodes.

Functional passivators are conventionally utilized in modifying the crystallization properties of perovskites to minimize the non-radiative recombination losses in perovskite light-emitting diodes (PeLEDs). However, the weak anchor ability of some commonly adopted molecules has limited passivation ability to perovskites and even may desorb from the passivated defects in a short period of time, which bring about plenty of challenges for further development of high-performance PeLEDs. Here, a multidentate molecule, formamidine sulfinic acid (FSA), is introduced as a novel passivator to perovskites. FSA has multifunctional groups (S≐O, C≐N and NH2 ) where the S≐O and C≐N groups enable coordination with the lead ions and the NH2 interacts with the bromide ions, thus providing the most effective chemical passivation for defects and in turn the formation of highly stable perovskite emitters. Moreover, the interaction between the FSA and octahedral [PbBr6 ]4- can inhibit the formation of unfavorable low-n domains to further minimize the inefficient energy transfer inside the perovskite emitters. Therefore, the FSA passivated green-emitting PeLED exhibits a high external quantum efficiency (EQE) of 26.5% with fourfold enhancement in operating lifetime as compared to the control device, consolidating that the multidentate molecule is a promising strategy to effectively and sustainably passivate the perovskites.

Toward a Method for Harmonized Susceptibility Testing of Mycoplasma bovis by Broth Microdilution.

Mycoplasma bovis is a fastidious pathogen of cattle causing massive economic losses in the calf and dairy industries worldwide. Since there is no approved standard method for antimicrobial susceptibility testing (AST) of M. bovis, the Clinical and Laboratory Standards Institute has requested the development of a suitable method. Therefore, this study aimed at developing a method for harmonized broth microdilution AST of M. bovis. For this, 131 M. bovis field isolates and M. bovis strain DSM 22781T were collected and macrorestriction analysis was performed to select 15 epidemiologically unrelated M. bovis strains for method validation steps. To select a suitable broth for AST of M. bovis, growth determinations were performed using five media and growth curves were compiled. Then, susceptibility testing was performed considering the exact (precondition of five identical MICs) and essential (MIC mode, accepting a deviation of ±1 dilution step) MIC agreements to evaluate the reproducibility of MIC values using a panel of 16 antimicrobial agents. Subsequently, the remaining field isolates were tested and the suitability of quality control (QC) strains was assessed. Growth experiments showed that SP4 broth was the only one of the five media that yielded sufficient growth of M. bovis. Therefore, it was selected as the test medium for AST and homogeneous MIC values were obtained (exact and essential agreements of 36 to 100% and 92 to 100%, respectively). For all other isolates tested, easy-to-read MIC endpoints were determined with this medium. High overall MIC50 and/or MIC90 values were observed for aminoglycosides and macrolides, and some isolates showed elevated MICs of fluoroquinolones, gentamicin, and/or tiamulin. Since the MICs of four commonly used QC strains were partially not within their ranges, a 20-fold MIC testing of M. bovis DSM 22781T was performed and met the criteria for a new QC strain. For harmonized AST of M. bovis, SP4 broth seems to be suitable with an incubation time of 72 ± 2 h and further validation of M. bovis DSM 22781T as a future QC strain is recommended.

Reproductive effects of pubertal exposure to neonicotinoid thiacloprid in immature male mice.

Thiacloprid (THIA) is a kind of neonicotinoid, a widely used insecticide class. Animal studies of adult and prenatal exposure to THIA have revealed deleterious effects on mammalian sperm fertility and embryonic development. A recent cross-sectional study linked higher THIA concentrations to delayed genitalia development stages in adolescent boys, suggesting that pubertal exposure to THIA may adversely affect reproductive development in immature males. Hence, this study aimed to investigate the effects of daily oral administration of THIA during puberty on the reproductive system of developing male mice. Young male C57 BL/6 J mice aged 21 days were administrated with THIA at concentrations of 10 (THIA-10), 50 (THIA-50) and 100 mg/kg (THIA-100) for 4 weeks by oral gavage. It is found that exposure to 100 mg/kg THIA diminished sexual behavior in immature male mice, caused a decrease in the spermatogenic cell layers and irregular arrangement of the seminiferous epithelium, and down-regulated the mRNA levels of spermatogenesis-related genes Ddx4, Scp3, Atg5, Crem, and Ki67, leading to an increase of sperm abnormality rate. In addition, THIA exposure at 50 and 100 mg/kg reduced the serum levels of testosterone and FSH, and decreased the expression levels of Star and Cyp11a1 related to testosterone biosynthesis. THIA exposure at 10 mg/kg did not produce any of the above significant changes. In conclusion, the high dose of THIA exposure impaired reproductive function in immature mice. It seems that THIA has no detrimental effects on the reproductive system of mice at low dose of 10 mg/kg.

Comparison of Dynamic Contrast-Enhanced MRI and Non-Mono-Exponential Model-Based Diffusion-Weighted Imaging for the Prediction of Prognostic Biomarkers and Molecular Subtypes of Breast Cancer Based on Radiomics.

BACKGROUND: Dynamic contrast-enhanced (DCE) MRI and non-mono-exponential model-based diffusion-weighted imaging (NME-DWI) that does not require contrast agent can both characterize breast cancer. However, which technique is superior remains unclear. PURPOSE: To compare the performances of DCE-MRI, NME-DWI and their combination as multiparametric MRI (MP-MRI) in the prediction of breast cancer prognostic biomarkers and molecular subtypes based on radiomics. STUDY TYPE: Prospective. POPULATION: A total of 477 female patients with 483 breast cancers (5-fold cross-validation: training/validation, 80%/20%). FIELD STRENGTH/SEQUENCE: A 3.0 T/DCE-MRI (6 dynamic frames) and NME-DWI (13 b values). ASSESSMENT: After data preprocessing, high-throughput features were extracted from each tumor volume of interest, and optimal features were selected using recursive feature elimination method. To identify ER+ vs. ER-, PR+ vs. PR-, HER2+ vs. HER2-, Ki-67+ vs. Ki-67-, luminal A/B vs. nonluminal A/B, and triple negative (TN) vs. non-TN, the following models were implemented: random forest, adaptive boosting, support vector machine, linear discriminant analysis, and logistic regression. STATISTICAL TESTS: Student's t, chi-square, and Fisher's exact tests were applied on clinical characteristics to confirm whether significant differences exist between different statuses (±) of prognostic biomarkers or molecular subtypes. The model performances were compared between the DCE-MRI, NME-DWI, and MP-MRI datasets using the area under the receiver-operating characteristic curve (AUC) and the DeLong test. P < 0.05 was considered significant. RESULTS: With few exceptions, no significant differences (P = 0.062-0.984) were observed in the AUCs of models for six classification tasks between the DCE-MRI (AUC = 0.62-0.87) and NME-DWI (AUC = 0.62-0.91) datasets, while the model performances on the two imaging datasets were significantly poorer than on the MP-MRI dataset (AUC = 0.68-0.93). Additionally, the random forest and adaptive boosting models (AUC = 0.62-0.93) outperformed other three models (AUC = 0.62-0.90). DATA CONCLUSION: NME-DWI was comparable with DCE-MRI in predictive performance and could be used as an alternative technique. Besides, MP-MRI demonstrated significantly higher AUCs than either DCE-MRI or NME-DWI. EVIDENCE LEVEL: 2. TECHNICAL EFFICACY: Stage 2.

Whole genome sequencing for the risk assessment of probiotic lactic acid bacteria.

Probiotic bacteria exhibit beneficial effects on human and/or animal health, and have been widely used in foods and fermented products for decades. Most probiotics consist of lactic acid bacteria (LAB), which are used in the production of various food products but have also been shown to have the ability to prevent certain diseases. With the expansion of applications for probiotic LAB, there is an increasing concern with regard to safety, as cases with adverse effects, i.e., severe infections, transfer of antimicrobial resistance genes, etc., can occur. Currently, in vitro assays remain the primary way to assess the properties of LAB. However, such methodologies are not meeting the needs of strain risk assessment on a high-throughput scale, in the context of the evolving concept of food safety. Analyzing the complete genetic information, including potential virulence genes and other determinants with a negative impact on health, allows for assessing the safe use of the product, for which whole-genome sequencing (WGS) of individual LAB strains can be employed. Genomic data can also be used to understand subtle differences in the strain level important for beneficial effects, or protect patents. Here, we propose that WGS-based bioinformatics analyses are an ideal and cost-effective approach for the initial in silico microbial risk evaluation, while the technique may also increase our understanding of LAB strains for food safety and probiotic property evaluation.

Overuse of food-grade disinfectants threatens a global spread of antimicrobial-resistant bacteria.

Food-grade disinfectants are extensively used for microbial decontamination of food processing equipment. In recent years, food-grade disinfectants have been increasingly used. However, the overuse of disinfectants causes another major issue, which is the emergence and spread of antimicrobial-resistant bacteria on a global scale. As the ongoing pandemic takes global attention, bacterial infections with antibiotic resistance are another ongoing pandemic that often goes unnoticed and will be the next real threat to humankind. Here, the effects of food-grade disinfectant overuse on the global emergence and spread of antimicrobial-resistant bacteria were reviewed. It was found that longtime exposure to the most common food-grade disinfectants promoted resistance to clinically important antibiotics in pathogenic bacteria, namely cross-resistance. Currently, the use of disinfectants is largely unregulated. The mechanisms of cross-resistance are regulated by intrinsic molecular mechanisms including efflux pumps, DNA repair system, modification of the molecular target, and metabolic adaptation. Cross-resistance can also be acquired by mobile genetic elements. Long-term exposure to disinfectants has an impact on the dissemination of antimicrobial resistance in soil, plants, animals, water, and human gut environments.

Application of problem-based self-designed experiments in physiology laboratory teaching.

A physiology laboratory course plays an important role in improving the scientific abilities of medical students. This study involved a teaching reform based on problem-based self-designed experiments in a physiology laboratory course. The study subjects were divided into two groups, i.e., students enrolled in 2019 were assigned to the traditional course control group (n = 146) and students enrolled in 2021 were assigned to the improved course test group (n = 128). Students in the test group were required to conduct self-designed experiments based on the questions for each experimental theme, in addition to completing the specified experimental items. At the end of the course, the differences in academic achievements between the two groups were compared. The results showed that compared to the control group, the students in the test group spent less time finishing the specified experimental items (P < 0.05). More students in the test group obtained good results in the operation assessment for the specified experiments (P < 0.05), and a significant increase in the number of winners in discipline-wise competitions, participants in scientific research projects, and academic publications was observed in the test group. Most of the students in the test group agreed that the self-designed experiment promoted their scientific thinking, helped them better understand theoretical knowledge, and improved their hands-on operation and team cooperation abilities. Our research showed that our teaching reform promoted students' self-directed learning and problem-solving abilities, stimulated their enthusiasm for scientific research, and was conducive to the cultivation of innovative medical talents.NEW & NOTEWORTHY This study involved a teaching reform based on problem-based self-designed experiments in a physiology laboratory course. Students in the test group were required to conduct self-designed experiments based on questions for each experimental theme, in addition to completing the specified experimental items. The results showed that the teaching reform promoted the students' self-directed learning and problem-solving ability, stimulated their enthusiasm for scientific research, and was conducive to cultivating innovative medical talents.

Colitis cystica profunda of the rectum diagnosed by endoscopic submucosal dissection.

Colitis cystica profunda is a rare and benign lesion characterized by mucus-containing cysts under the mucosa of the colon and rectum. We report a patient with localized colitis cystica profunda of the rectum diagnosed by endoscopic submucosal dissection. Although colitis cystica profunda is benign, it is sometimes indistinguishable from other malignant lesions. So early excision and biopsy make sense.

Proposal of a Method for Harmonized Broth Microdilution Antimicrobial Susceptibility Testing of Avibacterium gallinarum.

Avibacterium (Av.) gallinarum is an opportunistic pathogen in poultry, which, however, has also been associated with human disease. There is currently no approved method for antimicrobial susceptibility testing of this pathogen, so this study aimed at developing a harmonized broth microdilution method for Av. gallinarum that is suitable for diagnostic laboratories. For this, the Av. gallinarum CCUG 12391T type strain and 42 field isolates were collected and their species was confirmed by using a species-specific PCR assay and biochemical reactions. To select epidemiologically unrelated isolates, ApaI macrorestriction analysis was performed. Preliminary growth experiments were conducted with six culture media, and based on the results, four media were selected to compile growth curves with four isolates. Independent repetitions of MIC determinations were then performed to evaluate the reproducibility of the values. Cation-adjusted Mueller-Hinton broth (CAMHB) was initially selected as broth medium, but did not show sufficient homogeneity of MICs. Therefore, CAMHB plus 1% chicken serum and 0.0025% NADH was selected and showed a good homogeneity of MICs after 20 h and 24 h of incubation at 35 ± 2°C. This was reflected in essential MIC agreements ranging between 96% and 100%. Testing of a larger Av. gallinarum collection (n = 43) revealed that easily readable MICs could be obtained for the type strain and all isolates. Some Av. gallinarum showed elevated MICs of enrofloxacin (n = 35), nalidixic acid (n = 35), penicillin (n = 2), tetracycline (n = 19), and/or trimethoprim-sulfamethoxazole (n = 1). By using PCR analyses, the following antimicrobial resistance genes were detected: blaTEM, dfrA14, sul2, tet(B), tet(H). The study demonstrated that the proposed medium is suitable for a harmonized broth microdilution susceptibility testing of Av. gallinarum with a recommended incubation time of 20 to 24 h.

An Unsupervised Deep Learning Approach for Dynamic-Exponential Intravoxel Incoherent Motion MRI Modeling and Parameter Estimation in the Liver.

BACKGROUND: Dynamic-exponential intravoxel incoherent motion (IVIM) imaging is a potential technique for prediction, monitoring, and differential diagnosis of hepatic diseases, especially liver tumors. However, the use of such technique at voxel level is still limited. PURPOSE: To develop an unsupervised deep learning approach for voxel-wise dynamic-exponential IVIM modeling and parameter estimation in the liver. STUDY TYPE: Prospective. POPULATION: Ten healthy subjects (4 males; age 28 ± 6 years). FIELD STRENGTH/SEQUENCE: Single-shot spin-echo echo planar imaging (SE-EPI) sequence with monopolar diffusion-encoding gradients (12 b-values, 0-800 seconds/mm2 ) at 3.0 T. ASSESSMENT: The proposed deep neural network (DNN) was separately trained on simulated and in vivo hepatic IVIM datasets. The trained networks were compared to the approach combining least squares with Akaike information criterion (LSQ-AIC) in terms of dynamic-exponential modeling accuracy, inter-subject coefficients of variation (CVs), and fitting residuals on the simulated subsets and regions of interest (ROIs) in the left and right liver lobes. The ROIs were delineated by a radiologist (H.-X.Z.) with 7 years of experience in MRI reading. STATISTICAL TESTS: Comparisons between approaches were performed with a paired t-test (normality) or a Wilcoxon rank-sum test (nonnormality). P < 0.05 was considered statistically significant. RESULTS: In simulations, DNN gave significantly higher accuracy (91.6%-95.5%) for identification of bi-exponential decays with respect to LSQ-AIC (79.7%-86.8%). For tri-exponential identification, DNN was also superior to LSQ-AIC despite not reaching a significant level (P = 0.08). Additionally, DNN always yielded comparatively low root-mean-square error for estimated parameters. For the in vivo IVIM measurements, inter-subject CVs (0.011-0.150) of DNN were significantly smaller than those (0.049-0.573) of LSQ-AIC. Concerning fitting residuals, there was no significant difference between the two approaches (P = 0.56 and 0.76) in both the simulated and in vivo studies. DATA CONCLUSION: The proposed DNN is recommended for accurate and robust dynamic-exponential modeling and parameter estimation in hepatic IVIM imaging. LEVEL OF EVIDENCE: 2 TECHNICAL EFFICACY: Stage 1.

Short-Term Repeatability of in Vivo Cardiac Intravoxel Incoherent Motion Tensor Imaging in Healthy Human Volunteers.

BACKGROUND: Intravoxel incoherent motion (IVIM) tensor imaging is a promising technique for diagnosis and monitoring of cardiovascular diseases. Knowledge about measurement repeatability, however, remains limited. PURPOSE: To evaluate short-term repeatability of IVIM tensor imaging in normal in vivo human hearts. STUDY TYPE: Prospective. POPULATION: Ten healthy subjects without history of heart diseases. FIELD STRENGTH/SEQUENCE: Balanced steady-state free-precession cine sequence and single-shot spin-echo echo planar IVIM tensor imaging sequence (9 b-values, 0-400 seconds/mm2 and six diffusion-encoding directions) at 3.0 T. ASSESSMENT: Subjects were scanned twice with an interval of 15 minutes, leaving the scanner between studies. The signal-to-noise ratio (SNR) was evaluated in anterior, lateral, septal, and inferior segments of the left ventricle wall. Fractional anisotropy (FA), mean diffusivity (MD), mean fraction (MF), and helix angle (HA) in the four segments were independently measured by five radiologists. STATISTICAL TESTS: IVIM tensor indexes were compared between observers using a one-way analysis of variance or between scans using a paired t-test (normal data) or a Wilcoxon rank-sum test (non-normal data). Interobserver agreement and test-retest repeatability were assessed using the intraclass correlation coefficient (ICC), within-subject coefficient of variation (WCV), and Bland-Altman limits of agreements. RESULTS: SNR of inferior segment was significantly lower than the other three segments, and inferior segment was therefore excluded from repeatability analysis. Interobserver repeatability was excellent for all IVIM tensor indexes (ICC: 0.886-0.972; WCV: 0.62%-4.22%). Test-retest repeatability was excellent for MD of the self-diffusion tensor (D) and MF of the perfusion fraction tensor (fp ) (ICC: 0.803-0.888; WCV: 1.42%-9.51%) and moderate for FA and MD of the pseudo-diffusion tensor (D* ) (ICC: 0.487-0.532; WCV: 6.98%-10.89%). FA of D and fp and HA of D presented good test-retest repeatability (ICC: 0.732-0.788; WCV: 3.28%-8.71%). DATA CONCLUSION: The D and fp indexes exhibited satisfactory repeatability, but further efforts were needed to improve repeatability of D* indexes. LEVEL OF EVIDENCE: 2 TECHNICAL EFFICACY: Stage 1.

Development of a harmonized method for antimicrobial susceptibility testing of Bordetella avium using broth microdilution and detection of resistance genes.

AIMS: In response to a request from the Clinical and Laboratory Standards Institute (CLSI), the objective of this study was to develop a harmonized method for broth microdilution susceptibility testing of Bordetella (B.) avium, the major causative agent of infectious coryza in poultry. METHODS AND RESULTS: To find a suitable test medium, growth curves with four epidemiologically unrelated B. avium isolates were created in cation-adjusted Mueller-Hinton broth (CAMHB), CAMHB + 2.5% lysed horse blood and veterinary fastidious medium. All isolates showed good growth in CAMHB, therefore MIC values were determined using this medium and the homogeneity of the values was determined. An essential MIC agreement of 99.7% was calculated. Testing of a larger strain collection (n = 49) for their susceptibility to 24 antimicrobials confirmed the suitability of the tested method and revealed some isolates with elevated MICs of florfenicol (n = 1), streptomycin (n = 2), tetracyclines (n = 5), and trimethoprim/sulfamethoxazole (n = 6). PCR assays detected the resistance genes aadA1, dfrB1, floR, sul1, sul2 and tet(A). CONCLUSIONS: The method used enables easy reading and a good reproducibility of MIC values for B. avium. SIGNIFICANCE AND IMPACT OF STUDY: Application of the tested method allows harmonized resistance testing of B. avium and identification of isolates with elevated MIC values.

Clostridium butyricum alone or combined with 1, 25-dihydroxyvitamin D3 improved early-stage broiler health by modulating intestinal flora.

AIM: This study was conducted to investigate the effects of Clostridium butyricum in isolation or in combination with 1, 25-dihydroxyvitamin D3 in early-stage broilers. METHODS AND RESULTS: A total of 360 half male and half female Cobb broilers (1 day old) were randomly divided into four groups: Con (basal diet), Anti (basal diet+75 mg/kg chlortetracycline), Cb (basal diet+109  CFU per kg C. butyricum) and CD (basal diet+109  CFU per kg C. butyricum+25 µg/kg 1,25(OH)2 D3 ). The results were as follows: (1) Compared with Con, CD significantly increased ADG (p < 0.05). (2) Contrast with Con and Anti, Cb and CD significantly increased glutathione peroxidase and SOD in the serum and liver, and decreased malondialdehyde content in serum (p < 0.05). (3) In addition, the content of immunoglobulin (IgA, IgY and IgM) in Cb and CD birds was higher than that in Con birds (p < 0.05); the Cb supplementation decreased (p < 0.05) the contents of IL-8, IL-1ß and TNF-α than those in Con. (4) Cb and CD had lower caecal acetic and propionic content than the Anti group (p < 0.05). (5) The community richness of Con was significantly higher than that of Anti (p < 0.05). The relative abundance of Alistipes and Ruminococcaceae-UCG-014 in Cb and CD supplemented birds were lower than those in Con (p < 0.05). The relative abundant of Escherichia-Shigella in CD was higher than Con and Anti (p < 0.05). CONCLUSIONS: These data indicated that dietary C. butyricum and 1, 25-dihydroxyvitamin D3 can improve the growth performance, immunity responses, antioxidation, bone development and intestinal microflora in early-stage broilers. SIGNIFICANCE AND IMPACT OF THE STUDY: Oral administration of C. butyricum or C. butyricum combined with 1,25-dihydroxyvitamin D3 enhanced immunity and antioxidant activity in early-stage birds.

Establishment of enzyme-linked immunosorbent assays based on recombinant S1 and its truncated proteins for detection of PEDV IgA antibody.

Porcine epidemic diarrhea virus (PEDV) can infect pigs of all ages, especially piglets. PEDV has spread across Asia since the 1980s. The highly virulent variant PEDV broke out on a large scale and caused huge economic losses to the pig industry in late 2010 in China. Rapid detection methods with high specificity and sensitivity are urgently needed for the diagnosis and control of the disease. In this study, we divided the PEDV S1 gene into three segments and constructed the recombinant plasmids pFastBac1-S1T1 (aa 21-279), pFastBac1-S1T2 (aa 280-539) and pFastBac1-S1T3 (aa 540-788), which carry the different antigenic regions of the S1 gene. Truncated S1 proteins PEDV-S1T1/S1T2/S1T3 were obtained by a Bac-to-Bac expression system, with protein sizes of 36 kDa, 38 kDa and 38 kDa, respectively. Recombinant proteins presented high reactivity with the monoclonal antibody against PEDV and positive pig serum. Based on full-length S1 protein and these truncated proteins, we established indirect ELISA methods for the detection of PEDV IgA antibody. A total of 213 clinical serum samples were tested by the above indirect ELISA methods, and IFA was used as the gold standard. ROC curves revealed a significant correlation between S1-ELISA and S1T2-ELISA with a 0.9134 correlation coefficient and favourable sensitivity and specificity of S1-ELISA (93.24%, 95.68%) and S1T2-ELISA (89.33%, 94.16%). Our results also indicated that serum with higher neutralizing activity (SNT ≥ 40) had a higher IgA antibody level based on S1-ELISA, S1T1-ELISA and S1T2-ELISA. In conclusion, both S1-ELISA and S1T2-ELISA can be used as candidate systems for detecting anti-PEDV IgA antibody titers in serum, which can reflect the level of neutralizing activity in pigs after natural infection or vaccination. The above research results provide a basis for the prevention and control of PEDV and can be used in the detection of host anti-infective immunity and evaluation of vaccine immune effects.

Unsupervised monocular visual odometry via combining instance and RGB information.

Unsupervised deep learning methods have made significant progress in monocular visual odometry (VO) tasks. However, due to the complexity of the real-world scene, learning the camera ego-motion from the RGB information of monocular images in an unsupervised way is still challenging. Existing methods mainly learn motion from the original RGB information, lacking higher-level input from scene understanding. Hence, this paper proposes an unsupervised monocular VO framework that combines the instance and RGB information, named combined information based (CI-VO). The proposed method includes two stages. First is obtaining the instance maps of the monocular images, without finetuning on the VO dataset. Then we obtain the combined information from the two types of information, which is input into the proposed combined information based pose estimation network, named CI-PoseNet, to estimate the relative pose of the camera. To make better use of the two types of information, we propose a fusion feature extraction network to extract the fused features from the combined information. Experiments on the KITTI odometry and KITTI raw dataset show that the proposed method has good performance in the camera pose estimation task, which exceeds the existing mainstream methods.

Characterization of Two-Component System CitB Family in Salmonella Pullorum.

Salmonella enterica, serovar Gallinarum, biovar Pullorum, is an avian-specific pathogen which has caused considerable economic losses to the poultry industry worldwide. Two-component systems (TCSs) play an essential role in obtaining nutrients, detecting the presence of neighboring bacteria and regulating the expression of virulence factors. The genome analysis of S. Pullorum strain S06004 suggesting the carriage of 22 pairs of TCSs, which belong to five families named CitB, OmpR, NarL, Chemotaxis and LuxR. In the CitB family, three pairs of TCSs, namely CitA-CitB, DcuS-DcuR and DpiB-DpiA, remain unaddressed in S. Pullorum. To systematically investigate the function of the CitB family in S. Pullorum, four mutants, ΔcitAB (abbreviated as Δcit), ΔdcuSR (Δdcu), ΔdpiBA (Δdpi) and ΔcitABΔdcuSRΔdpiBA (Δ3), were made using the CRISPR/Cas9 system. The results demonstrated that the CitB family did not affect the growth of bacteria, the results of biochemical tests, invasion and proliferation in chicken macrophage HD-11 cells and the expression of fimbrial protein. But the mutants showed thicker biofilm formation, higher resistance to antimicrobial agents, enhanced tolerance to inhibition by egg albumen and increased virulence in chicken embryos. Moreover, the deletion of Dpi TCS was detrimental to survival after exposure to hyperosmotic and oxidative environments, as well as the long-term colonization of the small intestine of chickens. Collectively, we provided new knowledge regarding the possible role of the CitB family involved in the pathogenic processes of S. Pullorum.