C-type lectin 4 of Toxocara canis activates NF-ĸB and MAPK pathways by modulating NOD1/2 and RIP2 in murine macrophages in vitro.

Toxocara canis is a parasitic zoonose that is distributed worldwide and is one of the two pathogens causing toxocariasis. After infection, it causes serious public health and safety problems, which pose significant veterinary and medical challenges. To better understand the regulatory effects of T. canis infection on the host immune cells, murine macrophages (RAW264.7) were incubated with recombinant T. canis C-type lectin 4 (rTc-CTL-4) protein in vitro. The quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot were used to analyze the nucleotide-binding oligomerization domain-containing protein 1/2 (NOD1/2), receptor-interacting protein 2 (RIP2), nuclear factor kappa-light-chain enhancer of activated B cells (NF-κB), and mitogen-activated protein kinase (MAPK) on mRNA level and protein expression level in macrophages. Our results indicated that 10 µg/mL rTc-CTL-4 protein could modulate the expression of NOD1, NOD2, and RIP2 at both the transcriptional and translational levels. The protein translation levels of NF-κB, P-p65, p38, and P-p38 in macrophages were also modulated by rTc-CTL-4 protein. Macrophages were co-incubated with rTc-CTL-4 protein after siRNA silencing of NOD1, NOD2, and RIP2. The expression levels of NF-κB, P-p65, p38, and P-p38 were significantly changed compared with the negative control groups (Neg. Ctrl.). Taken together, rTc-CTL-4 protein seemed to act on NOD1/2-RIP2-NF-κB and MAPK signaling pathways in macrophages and might activate MAPK and NF-κB signaling pathways by regulating NOD1, NOD2, and RIP2. The insights from the above studies could contribute to our understanding of immune recognition and regulatory mechanisms of T. canis infection in the host animals.

3D-QSAR, molecular docking, and molecular dynamics analysis of novel biphenyl-substituted pyridone derivatives as potent HIV-1 NNRTIs.

When designing new medications targeting HIV-1, drug designers concentrate on reverse transcriptase (RT), the central enzyme of their concern. This is due to its vital role in converting single-stranded RNA into double-stranded DNA throughout the life cycle of HIV-1. In recent reports, a series of newly discovered pyridone derivatives with biphenyl substitutions have emerged as highly potent HIV-1 non-nucleoside reverse transcriptase inhibitors (NNRTIs), displaying impressive antiviral activity. To analyse the three-dimensional quantitative structure-activity relationship (3D-QSAR) of pyridone inhibitors with biphenyl substitutions, we employed CoMFA and CoMSIA methods in this study. The dataset comprises a total of 51 compounds. The findings of this research demonstrate that both the CoMFA (q2=0.688, r2=0.976, rpred2=0.831) and CoMSIA/SHE (q2=0.758, r2=0.968, rpred2=0.828) models exhibit excellent predictive capability and reliable estimation stability. According to the findings of the model, we designed a collection of eleven molecules that exhibit the potential for significantly improved predictive activity. We proceeded to investigate the binding patterns of these compounds to receptor proteins utilizing the molecular docking technique. To ensure the reliability of the docking results, we went on to validate them by conducting molecular dynamics simulations and performing accurate calculations of the binding free energy. Moreover, based on initial ADMET predictions, the results consistently indicate that the newly created molecule possesses favourable pharmacokinetic properties. This study will help to facilitate the development of efficient novel inhibitors that specifically target HIV-1's non-nucleoside reverse transcriptase (NNRTIs).Communicated by Ramaswamy H. Sarma.

Research on the Method of Counting Wheat Ears via Video Based on Improved YOLOv7 and DeepSort.

The number of wheat ears in a field is an important parameter for accurately estimating wheat yield. In a large field, however, it is hard to conduct an automated and accurate counting of wheat ears because of their density and mutual overlay. Unlike the majority of the studies conducted on deep learning-based methods that usually count wheat ears via a collection of static images, this paper proposes a counting method based directly on a UAV video multi-objective tracking method and better counting efficiency results. Firstly, we optimized the YOLOv7 model because the basis of the multi-target tracking algorithm is target detection. Simultaneously, the omni-dimensional dynamic convolution (ODConv) design was applied to the network structure to significantly improve the feature-extraction capability of the model, strengthen the interaction between dimensions, and improve the performance of the detection model. Furthermore, the global context network (GCNet) and coordinate attention (CA) mechanisms were adopted in the backbone network to implement the effective utilization of wheat features. Secondly, this study improved the DeepSort multi-objective tracking algorithm by replacing the DeepSort feature extractor with a modified ResNet network structure to achieve a better extraction of wheat-ear-feature information, and the constructed dataset was then trained for the re-identification of wheat ears. Finally, the improved DeepSort algorithm was used to calculate the number of different IDs that appear in the video, and an improved method based on YOLOv7 and DeepSort algorithms was then created to calculate the number of wheat ears in large fields. The results show that the mean average precision (mAP) of the improved YOLOv7 detection model is 2.5% higher than that of the original YOLOv7 model, reaching 96.2%. The multiple-object tracking accuracy (MOTA) of the improved YOLOv7-DeepSort model reached 75.4%. By verifying the number of wheat ears captured by the UAV method, it can be determined that the average value of an L1 loss is 4.2 and the accuracy rate is between 95 and 98%; thus, detection and tracking methods can be effectively performed, and the efficient counting of wheat ears can be achieved according to the ID value in the video.

Tracking spatio-temporal distribution and transmission of antibiotic resistance in aquatic environments by using ESBL-producing Escherichia coli as an indicator.

Wastewater treatment plants (WWTPs) play an important role in the production, and transmission of antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs) as a convergence for human, animal, and environmental wastewater. The aim of this study was to investigate the spatio-temporal variation and influencing factors of ARB in different functional areas of the urban WWTP and the connecting rivers for 1-year monitoring using extended-spectrum ß-lactamase-producing Escherichia coli (ESBL-Ec) as an indicator bacteria, and to study the transmission patterns of ARB in the aquatic environment. The results showed that ESBL-Ec isolates were identified from the WWTP (n = 219), including influent (n = 53), anaerobiotic tank (n = 40), aerobiotic tank (n = 36), activated sludge tank (n = 31), sludge thickner tank (n = 30), effluent (n = 16), and mudcake storage area (n = 13). The dehydration process can significantly remove the ESBL-Ec isolates; however, ESBL-Ec was still detected in samples collected from the effluent of the WWTP (37.0%). The detection rate of ESBL-Ec was significantly different across seasons (P < 0.05), and ambient temperature was negatively correlated with the detection rate of ESBL-Ec (P < 0.05). Furthermore, a high prevalence of ESBL-Ec isolates (29/187, 15.5%) was detected in samples collected from the river system. These findings emphasize that the high majority of ESBL-Ec in aquatic environments is alarming because it poses a significant threat to public health. Clonal transmission of ESBL-Ec isolates between the WWTP and rivers based on the spatio-temporal scale was observed by pulsed-field gel electrophoresis analysis, ST38 and ST69 ESBL-Ec clone were selected as prioritized isolates for antibiotic resistance monitoring in the aquatic environment. Further phylogenetic analysis showed human-associated (feces, blood) E. coli was the main source contributing to the presence of antibiotic resistance in aquatic environments. Longitudinal and targeted monitoring of ESBL-Ec in WWTPs and the development of effective wastewater disinfection strategies before effluent discharge from WWTPs are urgently required, to prevent and control the spread of antibiotic resistance in the environment.

Persistent transmission of carbapenem-resistant, hypervirulent Klebsiella pneumoniae between a hospital and urban aquatic environments.

The increasing prevalence of infections caused by carbapenem-resistant hypervirulent Klebsiella pneumoniae strains (CR-hvKP) prompts the question of whether these strains also circulate outside of clinical settings. However, the environmental occurrence and dissemination of CR-hvKP are poorly studied. In the current study, we investigated the epidemiological characteristics, and dissemination dynamics of carbapenem-resistant K. pneumoniae (CRKP) isolated from a hospital, an urban wastewater treatment plant (WWTP), and adjacent rivers in Eastern China during one year of monitoring. A total of 101 CRKP were isolated, 54 were determined to be CR-hvKP harboring pLVPK-like virulence plasmids, which were isolated from the hospital (29 out of 51), WWTP (23 out of 46), and rivers (2 out of 4), respectively. The period with lowest detection rate of CR-hvKP in the WWTP, August, corresponded with the lowest detection rate at the hospital. Comparing the inlet and outlet of the WWTP, no significant reduction of the detection of CR-hvKP and relative abundance of carbapenem resistance genes was observed. The detection rate of CR-hvKP and the relative abundance of carbapenemase genes were significantly higher in the WWTP in colder months compared to warmer months. Clonal dissemination of CR-hvKP clones of ST11-KL64 between the hospital and the aquatic environment, as well as the horizontal spread of IncFII-IncR and IncC plasmids carrying carbapenemase genes, was observed. Furthermore, phylogenetic analysis showed that the ST11-KL64 CR-hvKP strain has spread nationally by interregional transmission. These results indicated transmission of CR-hvKP clones between hospital and urban aquatic environments, prompting the need for improved wastewater disinfection and epidemiological models to predict the public health hazard from prevalence data of CR-hvKP.

Genomic insight into transmission mechanisms of carbapenem-producing Citrobacter spp. isolates between the WWTP and connecting rivers.

Carbapenem-resistant Enterobacteriaceae (CRE) poses major health risks worldwide. Most studies have focused on carbapenem resistance in Klebsiella pneumoniae and Escherichia coli; however, the occurrence and transmission of carbapenem-resistant Citrobacter spp. (CRCS) are poorly understood. In this study, we investigated the occurrence and potential transmission patterns of CRCS in different functional areas of an urban wastewater treatment plant (WWTP) and connecting rivers during one-year monitoring in Shandong Province, China. In total, 14 CRCS were detected in 376 environmental samples, including those from the WWTP inlet (n = 7), WWTP anaerobic tank (n = 2), and rivers (n = 5). Citrobacter braakii (n = 6) was the dominant subtype among 14 CRCS isolates, followed by Citrobacter freundii (n = 5), Citrobacter sedlakii (n = 2), and Citrobacter werkmanii (n = 1). All CRCS were resistant to imipenem, meropenem, ampicillin, amoxicillin/clavulanic acid, cefotaxime, ceftazidime, trimethoprim/sulfamethoxazole, and ciprofloxacin. Plasmid analysis showed that the blaKPC-2 gene was located on IncN and IncFII (Yp) plasmids, whereas the blaNDM gene was located on IncX3 and IncN2 plasmids. Clonal transmission of CRCS harboring carbapenem genes occurred between the WWTP and connecting rivers on a temporal or spatial scale. High genomic relatedness of NDM-5-producing C. sedlakii was identified between river water and WWTP aerosol, suggesting a potential exposure risk of CRCS for workers and surrounding residents near the WWTP. Furthermore, NDM-5-producing C. sedlakii isolated from rivers was related to C. sedlakii isolated from soil and well water in different regions of China, indicating that NDM-5-producing C. sedlakii may be widespread in China. These findings indicate that rare healthcare-associated pathogens such as CRCS can contribute to widespread carbapenem production in the environment; thus, CRCS should be continuously monitored.

Identification of muttony-related compounds in cooked mutton tallows and their flavor intensities subjected to phenolic extract from thyme (Thymus vulgaris L.).

Mutton possesses a typical flavor, known as "muttony" or "goaty", which significantly limits consumers' acceptability and its further popularization. Generally, this unpleasant flavor originates from mutton tallow. Thus, we first characterized the key volatiles of the cooked mutton tallow (CMT) via gas chromatography-mass spectrometry/olfactometry (GC-MS/O) and odor activity value (OAV). Combined with aroma recombination and omission tests, eleven compounds, involving 4-methyloctanoic acid, 4-methynonanoic acid, octanoic acid, decanoic acid, hexanal, heptanal, (E)-2-octenal, (E)-2-nonenal, (E)-2-decenal, 2-nonanone and 2-penty-furan, were screened out to be responsible for the "muttony" flavor. The objective of this study was to investigate the sensory property and acceptability of CMTs, elaborated with 4 different levels of thyme phenolic extract (TPE), through descriptive sensory analysis and key muttony-related compounds identification. The results showed that, of different TPEs employed, CMT plus TPE3 was the most effective strategy to control the key "muttony" contributors, thereby to improve flavor profile of CMT.

The shared NDM-positive strains in the hospital and connecting aquatic environments.

The spread of antibiotic-resistant priority pathogens outside hospital settings is, both, a significant public health concern and an environmental problem. In recent years, New Delhi Metallo-ß-lactamase (NDM)-positive strains have caused nosocomial infections with high mortality and poor prognosis worldwide. Our study investigated the links of NDM-positive strains between the hospital and the connecting river system in Jinan city, Eastern China by using NDM-producing Escherichia coli (NDM-EC) as an indicator via whole genome sequencing. Thirteen NDM-EC isolates were detected from 187 river water and sediment samples, while 9 isolates were identified from patients at the local hospital. All NDM-EC isolates were resistant to imipenem, meropenem, cefotaxime, cefoxitin, ampicillin, tetracycline, fosfomycin, piperacillin-tazobactam. The blaNDM-5 (n = 20) and blaNDM-9 (n = 2) genes were identified, which were predominantly on IncX3 plasmids (n = 13), followed by IncFII plasmids (n = 5) and IncFIA plasmids (n = 2). Conjugation experiments showed that 21 isolates could transfer NDM-harboring plasmids. The well-conserved blaNDM-5 genetic environment (ISAba125-blaNDM-5/9-bleMBL-trpF-dsbD-IS26) of these plasmids suggested a common genetic origin. Nine sequence types (STs) were detected, including three international high-risk clones ST167 (n = 8), ST410 (n = 1), and ST617 (n = 1). Phylogenetic analysis showed ST167 E. coli from the river was genotypically related to clinical isolates recovered from patients. Furthermore, ST167 isolates showed high genetic similarities with other clinical strains from geographically distinct regions. The genetic concordance between isolates from different sampling sites in the same river (ST218 clone), and different rivers (ST448 clone) raises concerns regarding the rapid dissemination of NDM-EC in the aquatic environment. The emergence and spread of the clinically relevant NDM-positive strains, especially for E. coli ST167 clone, an international high-risk clone associated with multi-resistance and virulence capacity, within and between the hospital and aquatic environments were elucidated, highlighting the need for attention and action.

Advances in the Formation and Control Methods of Undesirable Flavors in Fish.

Undesirable flavor formation in fish is a dynamic biological process, decreasing the overall flavor quality of fish products and impeding the sale of fresh fish. This review extensively summarizes chemical compounds contributing to undesirable flavors and their sources or formation. Specifically, hexanal, heptanal, nonanal, 1-octen-3-ol, 1-penten-3-ol, (E,E)-2,4-heptadienal, (E,E)-2,4-decadienal, trimethylamine, dimethyl sulfide, 2-methyl-butanol, etc., are characteristic compounds causing off-odors. These volatile compounds are mainly generated via enzymatic reactions, lipid autoxidation, environmentally derived reactions, and microbial actions. A brief description of progress in existing deodorization methods for controlling undesirable flavors in fish, e.g., proper fermenting, defatting, appropriate use of food additives, and packaging, is also presented. Lastly, we propose a developmental method regarding the multifunctional natural active substances made available during fish processing or packaging, which hold great potential in controlling undesirable flavors in fish due to their safety and efficiency in deodorization.