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This study aimed to investigate the differences in volatile composition and aroma of different fractions of Osmanthus fragrans absolute purified by molecular distillation. Gas chromatography-mass spectrometry (GC-MS) combined with orthogonal partial least-squares discrimination analysis (OPLS-DA), P-value, variable importance in the projection (VIP), and relative odor activity value (ROAV) were used for analysis. The light, heavy, and residue fractions of Osmanthus fragrans absolute were analyzed. A total of 73 volatile components belonging to 8 major categories were detected in the three fractions, including 22 alcohols, 5 aldehydes, 10 acids, 3 phenols, 5 ketones, 17 esters, 5 alkanes, and 6 olefins. Alcohols were the main contributors to the volatile components in each fraction, and the types and relative contents of volatile components significantly differed among the fractions (P1) of the three fractions, the key aroma contributing components (ROAV>1) of the 14 light fractions, 9 recombinant fractions, and 11 residue fractions were analyzed by the aroma profile analysis. Light fractions exhibited more intense floral, sweet, and pine woody aromas compared to recombinant and residue fractions. The light fractions had the highest types and relative contents of volatile components and showcased the most intense Osmanthus fragrans floral flavor. This study provided a theoretical basis for the comprehensive utilization of different fractions of Osmanthus fragrans absolute through molecular distillation using GC-MS combined with chemometrics methods.

An innovative chemometric method was developed to exploit visible and near-infrared (Vis-NIR) spectroscopy to guide food formulation to reach the anticipated and constant quality of final products. First, a total of 671 spectral variables related to the puree quality characteristics were identified by spectral variable selection methods. Second, the concentration profiles from multivariate curve resolution-alternative least squares (MCR-ALS) made it possible to reconstruct the identified spectral variables of formulated purees. Partial least square based on the reconstructed Vis-NIR spectral variables was evidenced to predict the final puree quality, such as a* values (RPD = 3.30), total sugars (RPD = 2.64), titratable acidity (RPD = 2.55) and malic acid (RPD = 2.67), based only on the spectral data of composed puree cultivars. These results open the possibility of controlling puree formulation: a multiparameter optimization of the color and taste of final puree products can be obtained using only the Vis-NIR spectral data of single-cultivar purees.

Vitamin C-based cluster therapy, which involves the combined application of hydrocortisone, vitamin C, and thiamine (HAT), is a recently proposed new treatment option for sepsis on top of conventional treatment. This therapy has a strong theoretical basis, but its clinical efficacy remains inconclusive. This review summarizes the rationale for HAT therapy for sepsis and describes the evaluation of its efficacy in clinical observational studies and randomized controlled trials, with the aim of providing a reference for the future clinical practice application of HAT therapy in sepsis.

Abstract Electric field is a powerful instrument in nanoscale engineering, providing wide functionalities for control in various optical and solid-state nanodevices. The development of a single optically resonant nanostructure operating with a charge-induced electrical field is challenging, but it could be extremely useful for novel nanophotonic horizons. Here, we show a resonant metal-semiconductor nanostructure with a static electric field created at the interface between its components by charge carriers generated via femtosecond laser irradiation. We study this field experimentally, probing it by second-harmonic generation signal, which, in our system, is time-dependent and has a non-quadratic signal/excitation power dependence. The developed numerical models reveal the influence of the optically induced static electric field on the second harmonic generation signal. We also show how metal work function and silicon surface defect density for different charge carrier concentrations affect the formation of this field. We estimate the value of optically-generated static electric field in this nanoantenna to achieve ≈108V/m. These findings pave the way for the creation of nanoantenna-based optical memory, programmable logic and neuromorphic devices.

Several animal models have been used to assist the development of vaccines and therapeutics since the COVID-19 outbreak. Due to the lack of binding affinity of mouse angiotensin-converting enzyme II (ACE2) to the S protein of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), increasing the susceptibility of mice to SARS-CoV-2 infection was considered in several ways. Here, we generated a COVID-19 mouse model expressing human ACE2 (hACE2) under the control of the CAG promoter. Overexpression of hACE2 did not pose a significant effect on weight growth. After SARS-CoV-2 inoculation, mice showed obvious viral replication and production of inflammation within 7 days, with a gradual decrease in body weight until death. Virological testing found that the virus can replicate in the respiratory system, small intestine, and brain. Additionally, this mouse model was applied to compare two antibody drug candidates, the anti-RBD antibody (MW06) and the mouse CD24-conjugated anti-RBD antibody (mCD24-MW06). Differences in antiviral effects between these two antibodies can be demonstrated in this mouse model when a challenge dose that invalidates the anti-RBD antibody treatment was used. This study provided a new mouse model for studying SARS-CoV-2 pathogenesis and evaluating potential interventions.

PurposeDespite strong efforts to promote human papillomavirus (HPV) vaccine and cervical cancer screening, cervical cancer remains a threat to women’s reproductive health. Some high-risk HPV types play a crucial role in the progression of cervical cancer and precancerous lesions. Therefore, HPV screening has become an important means to prevent, diagnose, and triage cervical cancer. This study aims to leverage artificial intelligence to predict individual risks of cervical intraepithelial neoplasia (CIN) in women with high-risk HPV infection and to recommend the appropriate triage strategy and follow-up period according to the risk level.Materials and methodsA total of 475 cases were collected in this study. The sources were from the Department of Gynecology and Obstetrics in a tertiary hospital, a case report on HPV from the PubMed website, and clinical data of cervical cancer patients from The Cancer Genome Atlas (TCGA) database. Through in-depth study of the interaction between high-risk HPV and its risk factors, the risk factor relationship diagram structure was constructed. A Classification of Lesion Stages (CLS) algorithm was designed to predict cervical lesion stages. The risk levels of patients were analyzed based on all risk factors, and follow-up periods were formulated for each risk level.ResultsOur proposed CLS algorithm predicted the probability of occurrence of CIN3—the precancerous lesion stage of cervical cancer. This prediction was based on patients’ HPV-16 and -18 infection status, age, presence of persistent infection, and HPV type. Follow-up periods of 3–6 months, 6–12 months, and 3- to 5-year intervals were suggested for high-risk, medium-risk, and low-risk patients, respectively.ConclusionA lesion prediction model was constructed to determine the probabilities of occurrence of CIN by analyzing individual data, such as patient lifestyle, physical assessments, and patient complaints, in order to identify high-risk patients. Furthermore, the potential implications of the calculated features were mined to devise prevention strategies.

IntroductionPrevious clinical investigations have reported inconsistent findings regarding the feasibility of utilizing indocyanine green fluorescence imaging (ICGFI) in laparoscopic liver tumor removal. This meta-analysis aims to comprehensively evaluate the safety and effectiveness of ICGFI in laparoscopic hepatectomy (LH).MethodsA systematic search of pertinent clinical studies published before January 30th, 2023 was conducted in databases including PubMed, Embase, Cochrane, and Web of Science. The search strategy encompassed key terms such as “indocyanine green fluorescence,” “ICG fluorescence,” “laparoscopic hepatectomy,” “hepatectomies,” “liver Neoplasms,” “hepatic cancer,” and “liver tumor.” Additionally, we scrutinized the reference lists of included articles to identify supplementary studies. we assessed the quality of the incorporated studies and extracted clinical data. Meta-analysis was performed using STATA v.17.0 software. Either a fixed-effects or a random-effects model was employed to compute combined effect sizes, accompanied by 95% confidence intervals (CIs), based on varying levels of heterogeneity.ResultsThis meta-analysis encompassed eleven retrospective cohort studies, involving 959 patients in total. Our findings revealed that, in comparison to conventional laparoscopic hepatectomy, patients receiving ICGFI-guided LH exhibited a higher R0 resection rate (OR: 3.96, 95% CI: 1.28, 12.25, I2 = 0.00%, P = 0.778) and a diminished incidence of intraoperative blood transfusion (OR: 0.42, 95% CI: 0.22, 0.81, I2 = 51.1%, P = 0.056). Additionally, they experienced shorter postoperative hospital stays (WMD: −1.07, 95% CI: −2.00, −0.14, I2 = 85.1%, P = 0.000). No statistically significant differences emerged between patients receiving ICGFI-guided LH vs. those undergoing conventional LH in terms of minimal margin width and postoperative complications.ConclusionICGFI-guided LH demonstrates marked superiority over conventional laparoscopic liver tumor resection in achieving R0 resection and reducing intraoperative blood transfusion rates. This technique appears to hold substantial promise. Nonetheless, further studies are needed to explore potential long-term benefits associated with patients undergoing ICGFI-guided LH.Systematic review registrationhttps://www.crd.york.ac.uk/prospero/, identifier CRD 42023398195.

The most important goal in cotton breeding is to increase the fiber yield which can be increased by improving a key trait, lint percentage (LP). However, narrow genetic background of upland cotton resulting in stagnant yield. In order to effectively dissect the genetic mechanism and enrich the gene loci of LP in cotton, the genetic diversity and population structure and a genome-wide association study (GWAS) was conducted using 204 upland cotton accessions based on 191 pairs of polymorphic microsatellite markers. The results were as follows: (1) The agronomic traits of the materials especially LP varied widely. (2) The SSRs revealed relatively high degree level of genetic diversity at DNA level. (3) Similarity coefficient cluster analysis of SSR markers indicated the influence of male parent on female parents was different after cross, the variation sites of agronomic traits could be further excavate in F1. (4) The GWAS revealed that 3 SSR markers were associated with LP: BNL3649-2, GH0132-2, CIR0246-3. There were two genes, Gh_D02G0125 and Gh_D11G2610, near the three SSR loci. These results provided a theoretical basis for improving fiber yield.

Protozoan parasites are a well-known threat to human health, particularly for people working at or visiting zoos, and potentially cause zoonotic diseases in humans. Captive wildlife may be potential reservoirs for human infection with protozoan parasites. Therefore, focusing on zoonotic protozoan infections in zoo animals is critical. However, there is no report on this topic in the Qinghai-Tibetan Plateau region. In this study, a total of 167 and 103 fecal samples were collected from 12 animal species from Qinghai-Tibet Plateau Wildlife Park in winter and summer, respectively, to detection the prevalence of infections and subtype distribution with Entamoeba sp., Cryptosporidium sp., Giardia duodenalis, Enteromicrosporidia bieneusi sp., Blastocystis sp. by PCR assay. The results showed that a total of 21 fecal samples collected in winter, including from 2 white-lipped deer, 8 Sika deer, 6 blue sheep, 2 wolves and 3 bears, were positive for Entamoeba, with a 12.6% (21/167) positive rate. However, 4.9% (5/103) of animals in summer were positive for Entamoeba, including 1 snow leopard, 1 tiger, 1 Tibetan argali and 2 mouflon. Moreover, 1 white-lipped deer and 1 bear were found to be positive for Blastocystis sp., one zoonotic STs (ST10) was identified and found in white-lipped deer. We found no effect on season on Blastocystis sp. and Entamoeba sp. colonization. To the best of our knowledge, this study is the first description of Blastocystis sp. and Entamoeba sp. infecting zoo animals in the plateau area. The findings provide the latest data on Entamoeba sp. and Blastocystis sp. in zoo animals in China.

ALBINO3 (ALB3) protein functions in the insertion and assembly of thylakoid membrane protein complexes and plays a critical role for chloroplast development in Arabidopsis. However, the biological function of ALB3 homologs in rice, OsALB3, remains elusive. Here, we identified a rice mutant, yellow leaf and lethal1 (yll1), that displayed yellow leaves and died at the seedling stage. The content of chlorophyll in yll1, compared with wild type, was significantly decreased. Transmission electron microscopy observation shows that the chloroplast of yll1 lacks thylakoid membranes. The causal mutation, which is located in OsALB3, was isolated by Mutmap+ combined with a simple mutation filtering process. Knockout of OsALB3 leads to yellow leaves and seedling lethality, mimicking the phenotype of yll1. OsALB3 is widely expressed and OsALB3 is chloroplast-localized. Moreover, the content of light-harvesting chlorophyll-binding proteins in yll1 is reduced. Together, our study demonstrated the essential role of OsALB3 in chloroplast development and provided clues to the possible conserved molecular function of ALB3 in rice.

Dermacentor nuttalli has been a focus of study because tick-borne pathogens have been widely identified in this tick from northern and southwestern China. The aim of this study was to characterize the life cycle of D. nuttalli under laboratory conditions and to detect spotted fever group (SFG) Rickettsia in the midgut and salivary glands of both field-collected and first laboratory generation adults. D. nuttalli ticks were collected in the field on the Qinghai-Tibetan Plateau from March to April 2021 and their life cycle was studied under laboratory conditions. Tick identify was molecularly confirmed, and SFG Rickettsia were detected in the midgut and salivary glands of males and females by PCR targeting different rickettsial genes. The results showed that the life cycle of D. nuttalli under laboratory conditions was completed in an average of 86.1 days. High positivity of Rickettsia spp. was detected in the midgut and salivary glands of both males (92.0%) and females (93.0%) of field-collected D. nuttalli ticks. However, a relatively lower positivity (4.0–6.0%) was detected in first laboratory generation adults. Furthermore, sequencing analysis showed that the Rickettsia sequences obtained in this study shared 98.6 to 100% nucleotide identity with Rickettsia slovaca and Rickettsia raoultii isolated from Dermacentor spp. in China. Phylogenetic analysis of Rickettsia spp. based on the gltA, ompA, ompB and sca4 genes revealed that the Rickettsia sequences obtained could be classified as belonging to R. slovaca and R. raoultii clades. This study described for the first time the life cycle of D. nuttalli from the Qinghai-Tibetan Plateau under laboratory conditions. Two species of SFG Rickettsia were detected in the midgut and salivary glands of males and females in both field-collected and first laboratory-generation adults of D. nuttalli. Our study provides new insights into pathogen detection in ticks in the Qinghai-Tibet Plateau, and the relationships among hosts, ticks, and pathogens.

Abstract Background Anaplasma, Babesia and Theileria are tick-borne pathogens (TBPs) that affect livestock worldwide. However, information on these pathogens in yaks (Bos grunniens) and Tibetan sheep (Ovis aries) on the Qinghai-Tibet Plateau (QTP), China, is limited. In this study, Anaplasma spp., Babesia spp. and Theileria spp. infections were assessed in yaks and Tibetan sheep from Qinghai Province. Methods A total of 734 blood samples were collected from 425 yaks and 309 Tibetan sheep at nine sampling sites. Standard or nested polymerase chain reaction was employed to screen all the blood samples using species- or genus-specific primers. Results The results showed that 14.1% (60/425) of yaks and 79.9% (247/309) of Tibetan sheep were infected with at least one pathogen. Anaplasma ovis, Anaplasma bovis, Anaplasma capra, Anaplasma phagocytophilum, Babesia bovis and Theileria spp. were detected in this study, with total infection rates for all the assessed animals of 22.1% (162/734), 16.3% (120/734), 23.6% (173/734), 8.2% (60/734), 2.7% (20/734) and 19.3% (142/734), respectively. For yaks, the infection rate of A. bovis was 6.4% (27/425), that of B. bovis was 4.7% (20/425) and that of Theileria spp. was 3.3% (14/425). Moreover, 52.4% (162/309) of the Tibetan sheep samples were infected with A. ovis, 30.1% (93/309) with A. bovis, 56.0% (173/309) with A. capra, 19.4% (60/309) with A. phagocytophilum and 41.4% (128/309) with Theileria spp. Conclusions This study revealed the prevalence of Anaplasma spp., Babesia spp. and Theileria spp. in yaks and Tibetan sheep in Qinghai Province, China, and provides new data for a better understanding of the epidemiology of TBPs in these animals in this area of the QTP, China. Graphical Abstract

Toxoplasmosis is a zoonotic disease caused by the obligate intracellular protozoan parasite T. gondii which is widely prevalent in humans and animals worldwide. The diagnosis of toxoplasmosis and distinguishing acute or chronic T. gondii infections have utmost importance for humans and animals. The TgSAG1, TgGRA7, and TgBAG1 proteins were used in the present study to develop the serological rSAG1-ELISA, rGRA7-ELISA and rBAG1-ELISA methods for the testing of T. gondii specific IgG and IgM antibodies and differentiating acute or chronic toxoplasmosis in 3733 animals, including Tibetan sheep, yaks, pigs, cows, cattle, horses, chickens, camels and donkeys from the Qinghai-Tibetan Plateau. The ELISA tests showed that the overall positivity of IgG antibody was 21.1% (786/3733), 15.3% (570/3733) and 18.2% (680/3733) for rSAG1-, rGRA7- and rBAG1-ELISA, respectively, and the positivity of IgM antibody was 11.8% (439/3733), 13.0% (486/3733) and 11.8% (442/3733) for rSAG1-, rGRA7- and rBAG1-ELISA, respectively. A total of 241 animals (6.5%) positive for all rSAG1-, rGRA7- and rBAG1-IgG were found in this study, and the 141 animals (3.8%) tested were anti-T. gondii IgM positive in all three ELISAs. Moreover, the 338, 284 and 377 animals were IgG positive in rSAG1 + rGRA7-, rBAG1 + rGRA7- and rSAG1 + rBAG1- ELISAs respectively, and the 346, 178 and 166 animals in rSAG1 + rGRA7-, rBAG1 + rGRA7- and rSAG1 + rBAG1-ELISAs were IgM positive respectively. The results confirmed that the application of SAG1, GRA7, and BAG1 recombinant antigens could successfully be used in the detection of specific IgG and IgM antibodies for distinguishing between acute or chronic T. gondii infections. It is inferred that the forms in which current animal species in the plateau area were infected with T. gondii, and the period of infection or the clinical manifestations of the current infections may be different. The present study provides substantial clinical evidence for the differential diagnosis of toxoplasmosis, and the classification of acute and chronic T. gondii infections.

Abstract Babesiosis is a tick-borne disease with global impact caused by parasites of the phylum Apicomplexa, genus Babesia. Typically, acute bovine babesiosis (BB) is characterized by fever, anemia, hemoglobinuria, and high mortality. Surviving animals remain persistently infected and become reservoirs for parasite transmission. Bovids in China can be infected by one or more Babesia species endemic to the country, including B. bovis, B. bigemina, B. orientalis, B. ovata, B. major, B. motasi, B. U sp. Kashi and B. venatorum. The latter may pose a zoonotic risk. Occurrence of this wide diversity of Babesia species in China may be due to a combination of favorable ecological factors, such as the presence of multiple tick vectors, including Rhipicephalus and Hyalomma, the coexistence of susceptible bovid species, such as domestic cattle, yaks, and water buffalo, and the lack of efficient measures of tick control. BB is currently widespread in several regions of the country and a limiting factor for cattle production. While some areas appear to have enzootic stability, others have considerable cattle mortality. Research is needed to devise solutions to the challenges posed by uncontrolled BB. Critical research gaps include risk assessment for cattle residing in endemic areas, understanding factors involved in endemic stability, evaluation of parasite diversity and pathogenicity of regional Babesia species, and estimation of whether and how BB should be controlled in China. Research should allow the design of comprehensive interventions to improve cattle production, diminish the risk of human infections, and increase the availability of affordable animal protein for human consumption in China and worldwide. In this review, we describe the current state of BB with reference to the diversity of hosts, vectors, and parasite species in China. We also discuss the unique risks and knowledge gaps that should be taken into consideration for future Babesia research and control strategies.

This study investigated the barley lesion mimic mutant (LMM) 5386, evidenced by a leaf brown spot phenotype localized on the chromosome 3H, and its conferred basal resistance to Fusarium graminearum. RNA-seq analysis identified 1453 genes that were differentially expressed in LMM 5386 compared to those in the wild type. GO and KEGG functional annotations suggested that lesion mimic formation was mediated by pathways involving oxidation reduction and glutathione metabolism. Additionally, reactive oxygen species (ROS) accumulation in brown spots was substantially higher in LMM 5386 than in the wild-type plant; therefore, antioxidant competence, which is indicated by ROS accumulation, was significantly lower in LMM 5386. Furthermore, the reduction of glycine in LMM 5386 inhibited glutathione biosynthesis. These results suggest that the decrease in antioxidant competence and glutathione biosynthesis caused considerable ROS accumulation, leading to programmed cell death, which eventually reduced the yield components in LMM 5386.

Neosporosis is a worldwide infectious disease caused by intracellular parasite Neospora caninum that is a major pathogen of abortion in cattle and neurological disorders in other hosts. However, limited data are available on animals exposed to N. caninum in the Qinghai-Tibetan Plateau Area (QTPA), and little is known about whether animals in the plateau area play an important role in the epidemiology of N. caninum. Therefore, indirect ELISAs based on a combination of NcSAG1 and NcGRA7 antigens were developed to examine both N. caninum-specific IgG and IgM antibodies in Tibetan sheep, yak, cow, pig, cattle, horse, chicken, camel, and donkey from the QTPA in this study. The results showed that all current species present- IgG and IgM-positive animals, and that the overall seroprevalence of N. caninum were 18.6 (703/3,782) and 48.1% (1,820/3,782) for the IgG and IgM antibodies, respectively. Further analysis found significant differences from different altitudes in IgG in Tibetan sheep and IgM in the yak. Hence, the present serological results indicate that the tested animal populations in the QTPA are suffering from N. caninum infections or have become carriers of N. caninum antibodies. To the best of our knowledge, this is the first report on current N. caninum-infected animals in the QTPA, the first epidemiology of neosporosis in cow and camel in China, and the first record of N. caninum IgM antibodies in all the surveyed animals in China. This study provides the latest valuable data on the epidemiology of neosporosis in China and in plateau areas of the world.

Abstract Direct and ultrasensitive detection of nanoscale objects is of great importance for materials science and biomedical application. Here, a printed single nanoparticle array‐based coloration biochip is developed for direct visualizing nanoscale objects with the microscopy image. Arising from the scattering cross section induced Mie scattering enhancement, the color change of nanoparticles with/without viruses can be observed with a conventional optical microscope. The scattering light from single nanoparticle has the size‐dependent characteristics of wavelength and intensity variation, allowing for optically detecting nanoscale virus. By comparing optical images before and after attaching viruses, quantitative detection can be achieved with the virus concentrations from 1.0 × 102 to 1.0 × 106 PFU mL–1 and a detection limit of 1.0 × 102 PFU mL–1. The antibody‐modified array can specifically recognize influenza viruses in 15 min. The result provides a promising approach to detect diverse biological samples, including protein, exosomes, and platelet microparticle.

Aim: To explore the relationship between the use of aspirin and the incidence of hepatocellular carcinoma (HCC).Methods: MEDLINE, EMBASE, Web of Science and Cochrane CENTRAL databases were searched systematically from the earliest available date to 13 March 2020. The primary outcome was incidence of HCC, and the secondary outcomes were recurrence and mortality of HCC. The results were expressed as the Hazard Ratio (HR) and 95% confidence interval (CI). Based on the heterogeneity evaluated with the I2 statistic, a meta-analysis was performed using either a random- or fixed-effects model.Results: A total of sixteen articles (2781100 participants) were included. There was lower incidence of HCC in aspirin users than those in non-aspirin users (HR, 0.56; 95% CI, 0.46-0.69; p < 0.001). Subgroup analysis further showed that the incidence of liver cancer in patients with alcoholic cirrhosis (HR, 0.14; 95% CI, 0.09-0.22; p < 0.001) and virus hepatitis (HR, 0.68; 95% CI, 0.62-0.74; p < 0.001) who use aspirin was lower than that of patients who do not use aspirin. In addition, aspirin was found to associate with decreased risk of HCC mortality (HR, 0.71; 95% CI, 0.65-0.78; p < 0.001), not HCC recurrence (HR, 0.52; 95% CI, 0.15-1.76; p = 0.291).Conclusions: Aspirin use is significantly associated with the low incidence rate of liver cancer.

Left bundle branch pacing (LBBP) is a physiological pacing technique that captures the left bundle branch (LBB) directly, causing the left ventricle (LV) to be excited earlier than the right ventricle (RV), resulting in a “iatrogenic” right bundle branch block (RBBB) pacing pattern. Several studies have recently shown that permanent LBBP can completely or partially narrow the wide QRS duration of the intrinsic RBBB in most patients with bradycardia, although the mechanisms by which this occurs has not been thoroughly investigated. This article presents a review of the LBBP in patients with intrinsic RBBB mentioned in current case reports and clinical studies, discussing the technique, possible mechanisms, future clinical explorations, and the feasibility of eliminating the interventricular dyssynchronization accompanied with LBBP.

BackgroundNeutrophil elastase (NE) is associated with sepsis occurrence and progression. We hypothesized that the NE inhibitor Sivelestat might modulate abnormal gut microbiota and metabolites during sepsis.MethodsSixty Sprague-Dawley (SD) rats were randomly divided into sham control (SC), sepsis (CLP), and sepsis+Sivelestat (Sive) groups. The rats’ survival status was monitored for 24 hours postoperatively, and feces were collected for microbiome and non-targeted metabolomics analyses.ResultsSivelestat administration significantly improved the survival of septic rats (80% vs 50%, P = 0.047). Microbiome analysis showed that the microbiota composition of rats in the CLP group was significantly disturbed, as potential pathogens such as Escherichia-Shigella and Gammaproteobacteria became dominant, and the beneficial microbiota represented by Lactobacillus decreased. These changes were reversed in Sive group, and the overall microbial status was restored to a similar composition to SC group. Differential analysis identified 36 differential operational taxonomic units and 11 metabolites between the Sive and CLP groups, such as 6-Aminopenicillanic acid, gamma-Glutamyl-leucine, and cortisone (variable importance in projection>1and P

Abstract Objective: To observe the effects of preoperative right stellate ganglion block on perioperative atrial fibrillation in patients undergoing lung lobectomy. Methods: Two hundred patients who underwent a scheduled lobectomy were randomly divided into the S and C groups. The S group was injected with 4 mL of 0.2% ropivacaine under ultrasound guidance, and the C group did not receive stellate ganglion block. The patients underwent continuous ECG monitoring, and the incidences of atrial fibrillation and other types of arrhythmias were recorded from the start of surgery to 24 hours after surgery. Results: The respective incidences of atrial fibrillation in the S group and the C group were 3% and 10% (p = 0.045); other atrial arrhythmias were 20% and 38% (p = 0.005); and ventricular arrhythmia were 28% and 39% (p = 0.09). Conclusions: The results of the study indicated that preoperative right stellate ganglion block can effectively reduce the incidence of intraoperative and postoperative atrial fibrillation.

The wayside acoustic defective bearing detector system (TADS) is located on both sides of the railway, so that the acoustic signals recorded by the microphone not only include the sound from the train bearings but also include it from the other disturbance sources. The heavy noise and multisource acoustic signals would badly reduce the reliability and accuracy of the detection result of the TADS. In order to extract the useful information from the recorded signal exactly and efficiently, a novel denoising method based on the Short-time Fourier transform (STFT) and improved Crazy Climber algorithm was improved in this paper. Firstly, the STFT was performed on the recorded acoustic signals in order to obtain the time-frequency distribution matrix. Based on the original algorithm, the novel movement rule and the fitting process of the ridge lines were presented which could extract the time-frequency ridge lines of the acoustic signal accurately and rapidly. In this way, the important information from the train bearings could be divided from the heavy noise and other signals. Finally, the simulation and experimental verifications were carried out, and the denoising method based on the STFT and improved Crazy Climber algorithm has proved to be effective in extracting ridge lines of the time-frequency distribution matrix and dividing the useful information form the recorded acoustic signals.

Background: Although numerous studies are conducted every year on how to reduce the fatality rate associated with sepsis, it is still a major challenge faced by patients, clinicians, and medical systems worldwide. Early identification and prediction of patients at risk of sepsis and adverse outcomes associated with sepsis are critical. We aimed to develop an artificial intelligence algorithm that can predict sepsis early.Methods: This was a secondary analysis of an observational cohort study from the Intensive Care Unit of the First Affiliated Hospital of Zhengzhou University. A total of 4,449 infected patients were randomly assigned to the development and validation data set at a ratio of 4:1. After extracting electronic medical record data, a set of 55 features (variables) was calculated and passed to the random forest algorithm to predict the onset of sepsis.Results: The pre-procedure clinical variables were used to build a prediction model from the training data set using the random forest machine learning method; a 5-fold cross-validation was used to evaluate the prediction accuracy of the model. Finally, we tested the model using the validation data set. The area obtained by the model under the receiver operating characteristic (ROC) curve (AUC) was 0.91, the sensitivity was 87%, and the specificity was 89%.Conclusions: This newly established machine learning-based model has shown good predictive ability in Chinese sepsis patients. External validation studies are necessary to confirm the universality of our method in the population and treatment practice.

Gibberella stalk rot caused by Fusarium graminearum is one of the devastating diseases of maize that causes significant yield losses worldwide. The molecular mechanisms regulating defense against this pathogen remain poorly understood. According to recent studies, a major oxylipin hormone produced by 13-lipoxygenases (LOX) namely jasmonic acid (JA) has been associated with maize susceptibility to GSR. However, the specific roles of numerous 9-LOX-derived oxylipins in defense against Gibberella stalk rot (GSR) remain unexplained. In this study, we have shown that disruption of a 9-LOX gene, ZmLOX5, resulted in increased susceptibility to GSR, indicating its role in defense. To understand how ZmLOX5 regulates GSR resistance, we conducted transcriptome and oxylipin profiling using a zmlox5-3 mutant and near-isogenic wild type B73, upon infection with F. graminearum. The results showed that JA biosynthetic pathway genes were highly up-regulated, whereas multiple 9-LOX pathway genes were down-regulated in the infected zmlox5-3 mutant. Furthermore, oxylipin profiling of the mutant revealed significantly higher contents of several jasmonates but relatively lower levels of 9-oxylipins in zmlox5-3 upon infection. In contrast, B73 and W438, a more resistant inbred line, displayed relatively lower levels of JAs, but a considerable increase of 9-oxylipins. These results suggest antagonistic interaction between 9-oxylipins and JAs, wherein 9-oxylipins contribute to resistance while JAs facilitate susceptibility to F. graminearum.

Tick-borne diseases (TBDs) can cause serious economic losses and are very important to animal and public health. To date, research on TBDs has been limited in Qinghai-Tibet Plateau, China. This epidemiological investigation was conducted to evaluate the distribution and risk factors of Anaplasma spp. and Ehrlichia chaffeensis in livestock in Qinghai. A total of 566 blood samples, including 330 yaks (Bos grunniens) and 236 Tibetan sheep (Ovis aries) were screened. Results showed that A. bovis (33.3%, 110/330) and A. phagocytophilum (29.4%, 97/330) were most prevalent in yaks, followed by A. ovis (1.2%, 4/330), A. capra (0.6%, 2/330), and E. chaffeensis (0.6%, 2/330). While A. ovis (80.9%, 191/236) and A. bovis (5.1%, 12/236) infection was identified in Tibetan sheep. To our knowledge, it is the first time that A. capra and E. chaffeensis have been detected in yaks in China. Apart from that, we also found that co-infection of A. bovis and A. phagocytophilum is common in yaks (28.2%, 93/330). For triple co-infection, two yaks were infected with A. bovis, A. phagocytophilum, and A. capra, and two yaks were infected with A. bovis, A. phagocytophilum, and E. chaffeensis. Risk analysis shows that infection with A. bovis, A. phagocytophilum, and A. ovis was related to region and altitude. This study provides new data on the prevalence of Anaplasma spp. and E. chaffeensis in Qinghai, China, which may help to develop new strategies for active responding to these pathogens.

As one of the important parameters of bearing operation, temperature is a key metric to diagnose the state of service of a bearing. However, there are still some shortcomings in the study of the temperature variation law for damaged bearings. In this paper, according to the structural characteristics of bearings, the influence law of thermal-solid coupling effect on bearing structure is considered, and a novel transient temperature analysis model of damaged bearings is established. First, a quasi-static analysis of the bearing is performed to obtain the variation laws of the key parameters of the bearing under thermal expansion. Then, the load variation law of the bearing under the condition of damage is discussed, and the heat generation and heat transfer of the damaged bearing during operation are studied. Based on the thermal grid method, a transient temperature analysis model of the damaged bearing is developed. Finally, the model is tested experimentally and the influence of the rotate speed and load on the bearing temperature variation is analyzed. The results show that the established model can effectively predict the temperature variation and thermal equilibrium temperature of damaged bearings.

Anaplasma genus infects the blood cells of humans and animals by biting, causing zoonotic anaplasmosis. However, limited data are available on carrier animals for Anaplasma spp. antibodies in the Qinghai–Tibetan Plateau Area. Therefore, a serological indirect ELISA diagnostic method based on the major surface protein 5 (MSP5), derived from Anaplasma phagocytophilum, was developed in this study to analyze both IgG and IgM antibodies of Anaplasma spp. in a total of 3952 animals from the Qinghai–Tibetan Plateau, including yaks (Bos grunniens), cows (Bos taurus), cattle (Bos taurus domesticus), Tibetan sheep (Ovis aries), horses (Equus ferus caballus), pigs (Sus domesticus), chickens (Gallus gallus domesticus), donkeys (Equus asinus), stray dogs (Canis sp.), and stray cats (Felis sp.). The results showed that recombinant MSP5 protein was expressed and was successfully used to establish the indirect ELISA methods. The overall positivity for Anaplasma IgG and IgM antibodies was 14.6% (578/3952) and 7.9% (312/3952), respectively, and a total of 123 animals (3.1%) were both IgG- and IgM-positive. Moreover, the most prevalent Anaplasma IgG positivity was exhibited by donkeys (82.5%), followed by stray dogs, Tibetan sheep, pigs, chickens, horses, yaks, cows, cattle, and stray cats. The analysis for IgM antibody positivity revealed that IgM positivity was the most prevalent in the stray dogs (30.1%), followed by horses, yaks, Tibetan sheep, cows, stray cats, and cattle. Moreover, the results revealed significant differences (p < 0.05) at different altitudes in Anaplasma-specific IgG in the yaks, Tibetan sheep, and horses, and in IgM in the yaks and Tibetan sheep. In conclusion, this study is the first to demonstrate that yaks, cows, cattle, Tibetan sheep, horses, donkeys, stray dogs, stray cats, pigs, and chickens living in the Qinghai–Tibet Plateau are carrier animals for Anaplasma spp. IgG or IgM antibodies. The current findings provide valuable current data on the seroepidemiology of anaplasmosis in China and for plateau areas of the world.

Four supported α-diimine nickel(II) catalysts covalently linked to silica via hydroxyl functionality on α-diimine acenaphthequinone-backbone were prepared and used in slurry polymerizations of ethylene to produce branched polyethylenes. The catalytic activities of these still reached 106 g/molNi·h at 70 °C. The life of the supported catalyst is prolonged, as can be seen from the kinetic profile. The molecular weight of the polyethylene obtained by the 955 silica gel supported catalyst was higher than that obtained by the 2408D silica gel supported catalyst. The melting points of polyethylene obtained by the supported catalysts S-C1-a/b are all above 110 °C. Compared with the homogeneous catalyst, the branching numbers of the polyethylenes obtained by the supported catalysts S-C1-a/b is significantly lower. The polyethylenes obtained by supported catalyst S-C1-a/b at 30–50 °C are free-flowing particles, which is obviously better than the rubber-like cluster polymer obtained from homogeneous catalyst.

Abstract Background The thrombospondin-related anonymous protein (TRAP) was first discovered in the sporozoite of Plasmodium falciparum and TRAP family proteins are secreted by micronemes and transported to the parasite surface to participate in the invasion process. Various TRAP proteins have been identified in apicomplexan protozoans, but there have been few reports about TRAP proteins in Babesia orientalis. Methods The functional domain of TRAP2 in B. orientalis was cloned, sequenced, characterized and compared to the TRAP sequences of related apicomplexan parasites. The functional domain of BoTRAP2 was truncated, named BoTRAP2-1, and then cloned into the pET-28a expression vector. Rabbit anti-rBoTRAP2-1 polyclonal antibody was produced by immunizing three rabbits. Western blot analysis was used to identify the native form and immunogenicity of BoTRAP2. The localization of BoTRAP2 was identified by indirect fluorescence assay (IFA). Results The amplified genes of BoTRAP2 are 2817 bp in length, encoding a functional domain of about 938 aa with two vWFA domains, one TSP domain and one transmembrane domain. The amino acid sequence of BoTRAP2 has a high similarity with that of B. bovis and B. gibsoni. The predicted tertiary structure of truncated BoTRAP2-1 confirmed that BoTRAP2 contains two vWFA domains and a TSP domain, the main functional areas of the protein. The native BoTRAP2 was identified from B. orientalis lysate by using rabbit polyclonal anti-rBoTRAP2-1. A band corresponding to rBoTRAP2-1 was detected by reaction with serum from a B. orientalis-infected water buffalo, indicating that the protein has a high immunogenicity. IFA showed that BoTRAP2 is mainly localized on the apical end of parasites by rabbit anti-rBoTRAP2-1 polyclonal serum. Conclusions The rBoTRAP2 could differentiate serum from B. orientalis-infected water buffalo and normal water buffalo, implicating that BoTRAP2 has high immunogenicity and could serve as a candidate antigen for diagnosis of B. orientalis infection in buffalo.

Abstract Background The thrombospondin-related anonymous protein (TRAP) family, a kind of transmembrane protein, is widely distributed with a conserved feature of structure in all apicomplexan parasites and plays a crucial role in the gliding motility and survival of parasites. Methods The Babesia orientalis TRAP1 gene (BoTRAP1) was truncated and cloned into a pET-42b expression vector and expressed as a GST-tag fusion protein with a TEV protease site. Rabbit anti-rBoTRAP1 antibody was produced and purified using a protein A chromatography column. Western blot analysis was performed to identify the native protein of BoTRAP1 and differentiate B. orientalis-infected positive from negative serum samples. The localization of BoTRAP1 on merozoites was identified by the indirect florescent antibody test (IFAT). Results The partial sequence of the TRAP1 gene was cloned from B. orientalis cDNA and identified to contain a von Willebrand factor A (vWFA) region and a thrombospondin type-1 (TSP-1) domain; it had a length of 762 bp, encoding a polypeptide of 254 amino acid residues with a predicted size of 28.2 kDa. The partial sequence was cloned into a pET-42b expression vector and expressed in E. coli as a GST fusion protein. Western blot indicated that rBoTRAP1 has a high immunogenicity and can differentiate B. orientalis-infected positive and negative serum samples collected from water buffaloes. IFAT showed that BoTRAP1 is mainly localized on the apical end of intracellular parasites by using polyclonal antibodies (PcAb) against rBoTRAP1. Meanwhile, the PcAb test also identified the native BoTRAP1 as a ~65 kDa band from B. orientalis lysates. The predicted 3D structure of BoTRAP1 contains a metalion-dependent adhesion site (MIDAS), which could be important for interaction with ligand on the surface of the host cells. Conclusions Like all known protozoa, B. orientalis has a TRAP family, comprising TRAP1, TRAP2, TRAP3 and TRAP4. The newly identified and characterized BoTRAP1 may play a key role in the invasion of B. orientalis into water buffalo erythrocytes.

Gibberella stalk rot (GSR) caused by Fusarium graminearum is one of the most devastating diseases causing significant yield loss of maize, and GSR resistance is a quantitative trait controlled by multiple genes. Although a few quantitative trait loci/resistance genes have been identified, the molecular mechanisms underlying GSR resistance remain largely unexplored. To identify potential resistance genes and to better understand the molecular mechanism of GSR resistance, a joint analysis using a comparative transcriptomic and metabolomic approaches was conducted using two inbred lines with contrasting GSR resistance, K09 (resistant) and A08 (susceptible), upon infection with F. graminearum. While a substantial number of differentially expressed genes associated with various defense-related signaling pathways were identified between two lines, multiple hub genes likely associated with GSR resistance were pinpointed using Weighted Gene Correlation Network Analysis and K-means clustering. Moreover, a core set of metabolites, including anthocyanins, associated with the hub genes was determined. Among the complex co-expression networks, ZmHIR3 showed strong correlation with multiple key genes, and genetic and histological studies showed that zmhir3 mutant is more susceptible to GSR, accompanied by enhanced cell death in the stem in response to infection with F. graminearum. Taken together, our study identified differentially expressed key genes and metabolites, as well as co-expression networks associated with distinct infection stages of F. graminearum. Moreover, ZmHIR3 likely plays a positive role in disease resistance to GSR, probably through the transcriptional regulation of key genes, functional metabolites, and the control of cell death.

Babesia orientalis, a major infectious agent of water buffalo hemolytic babesiosis, is transmitted by Rhipicephalus haemaphysaloides. However, no effective vaccine is available. Essential antigens that are involved in parasite invasion of host red blood cells (RBCs) are potential vaccine candidates. Therefore, the identification and the conduction of functional studies of essential antigens are highly desirable. Here, we evaluated the function of B. orientalis merozoite surface antigen 2c1 (BoMSA-2c1), which belongs to the variable merozoite surface antigen (VMSA) family in B. orientalis. We developed a polyclonal antiserum against the purified recombinant (r)BoMSA-2c1 protein. Immunofluorescence staining results showed that BoMSA-2c1 was expressed only on extracellular merozoites, whereas the antigen was undetectable in intracellular parasites. RBC binding assays suggested that BoMSA-2c1 specifically bound to buffalo erythrocytes. Cytoadherence assays using a eukaryotic expression system in vitro further verified the binding and inhibitory ability of BoMSA-2c1. We found that BoMSA-2c1 with a GPI domain was expressed on the surface of HEK293T cells that bound to water buffalo RBCs, and that the anti-rBoMSA2c1 antibody inhibited this binding. These results indicated that BoMSA-2c1 was involved in mediating initial binding to host erythrocytes of B. orientalis. Identification of the occurrence of binding early in the invasion process may facilitate understanding of the growth characteristics, and may help in formulating strategies for the prevention and control of this parasite.

Lectin receptor-like kinases (LecRLKs), a plant-specific receptor-like kinase (RLK) sub-family, have been recently found to play crucial roles in plant development and responses to abiotic and biotic stresses. In this review, we first describe the classification and structures of Lectin RLKs. Then we focus on the analysis of functions of LecRLKs in various biological processes and discuss the status of LecRLKs from the ligands they recognize, substrate they target, signaling pathways they are involved in, to the overall regulation of growth-defense tradeoffs. LecRLKs and the signaling components they interact with constitute recognition and protection systems at the plant cell surface contributing to the detection of environmental changes monitoring plant fitness.

Toxoplasma gondii and Neospora caninum belong to the Apicomplexan protozoa which is an obligate intracellular parasite, causing toxoplasmosis and neosporosis throughout the world. Cats and dogs are the definitive hosts of these two parasites. However, information on the epidemiology of toxoplasmosis and neosporosis in stray cats and dogs in the Qinghai–Tibetan Plateau Area (QTPA) is limited, and little is known about the diversity of the diseases. Therefore, the aim of this study was to perform indirect ELISA tests based on recombinant TgSAG1, TgGRA1, NcSAG1 and NcGRA7 proteins to establish a detailed record of the seroprevalence of T. gondii and N. caninum-specific IgG and IgM antibodies in serum samples and to develop qPCR amplification based on TgB1 and NcNc5 genes to conduct molecular epidemiology in feces from stray cats and dogs in the QTPA. In the current study, a total of 128 cat serum samples were analyzed through serological tests in which 53 (41.4%) and 57 (44.5%) samples were found positive for T. gondii specific-IgG and IgM antibodies, and 2 (1.6%) and 74 (57.8%) samples were confirmed positive for N. caninum specific-IgG and IgM antibodies, respectively. Out of 224 stray dog sera, 59.8% and 58.9% were recorded as positive against anti-Toxoplasma IgG and IgM antibodies, 17.9% and 64.7% were detected positive against Neospora IgG and IgM. On the other hand, 1 of 18 cat fecal samples was successfully amplified within the Ct value of 10 to 30 while no cat was positive for neosporosis. Moreover, a higher prevalence of toxoplasmosis in stray dogs (14.5%, 16/110) than of neosporosis (5.5%, 6/110) with different parasite numbers were found. Further analysis showed that no significant sex differences were found nor between the overall infection rates of T. gondii and N. caninum in this study. This study suggests that stray cats and dogs play key roles in the transmission and prevalence of T. gondii and N. caninum in the plateau area.

Background: Bai-Hu-Tang (BHT), a Chinese herbal decoction used as an antipyretic agent, results from the combination of Anemarrhena asphodeloides Bunge, Glycyrrhizae, Japonica rice, and Gypsum. In our previous study, we identified nanoaggregates in BHT. However, the present study aimed to analyze and elucidate the mechanism of nanoaggregate formation and to investigate its antipyretic effect. Methods: A BHT decoction extract was split into 15 groups, and in each group, the extract was further separated into two solutions: Nano-phase and Decoction. The physicochemical properties of these solutions, such as particle size, salinity, conductivity, and surface tension were investigated, and analyzed the 15 groups of by transmission electron microscopy (TEM) and fingerprint chromatography. Furthermore, the antipyretic effect of nanoaggregates was evaluated through enzyme-linked immunosorbent assays, HE staining, Western Blot, and Real-time PCR. Results: In the 15 groups, the salinity and conductivity results showed a promoting and stabilizing effect towards the Nano-phase formation. Analysis of the surface tension indicated good solubilization of Radix Glycyrrhizae. The TEM analysis of the BHT separated extracts revealed that only in the presence of Japonica rice the Nano-phase is formed. Sixteen common peaks were identified in the BHT fingerprint chromatogram, and the main chemical components were Neomangiferin, Mangiferin, Liquiritin, and Ammonium glycyrrhizinate. Furthermore, BHT and nanoaggregates from Bai-Hu-Tang (N-BHT) groups did not differ in the main chemical components. Additionally, the N-BHT group had the same antipyretic effect compared with the BHT group. However, the pathological analysis indicated that treatment with N-BHT could ameliorate the lung damage in the rat. At the same time, N-BHT group inhibited expression of several proteins, specifically IL-1β, TRPV4, NF-κB, and TNF-α, which agreed with the Real-time PCR results. Conclusion: We identified the key factors that are involved in the nano-phase formation. Also, by Western blot and Real-time PCR methods, we investigated the N-BHT mechanism of antipyretic action. The discovery of the N-BHT formation would provide a new idea of studying traditional Chinese medicine decoction.

Human babesiosis is caused by apicomplexan Babesia parasites, including Babesia microti, Babesia crassa, Babesia sp. MOI, Babesia divergens, Babesia duncani, and Babesia venatorum. Among them, B. microti is the most common cause of human and rodent babesiosis. Currently, no vaccine is available, and drugs for the treatment have high failure rates and side effects. Due to lack of a traditional tricarboxylic acid cycle (TCA cycle) and its dominant dependence on anaerobic metabolism to produce ATP, B. microti lactate dehydrogenase (BmLDH) was assumed to play a critical role in B. microti ATP supply. Our previous study demonstrated that BmLDH is a potential drug target and Arg99 is a crucial site. Herein, a molecular docking was performed based on the crystal structure of BmLDH from a series of gossypol derivatives or structural analogs to find the potent inhibitors interacting with the residue Arg99, and three naphthalene-based compounds 2,6-naphthalenedicarboxylic acid (NDCA), 1,6-dibromo-2-hydroxynapthalene 3-carboxylic acid (DBHCA), and 3,5-dihydroxy 2-napthoic acid (DHNA) were selected for further tests. Enzyme activity inhibitory experiments show that DBHCA and DHNA inhibit recombinant BmLDH (rBmLDH) catalysis with ~109-fold and ~5,000-fold selectivity over human LDH, respectively. Surface plasmon resonance (SPR) assays demonstrate that DHNA has a lower KD value to BmLDH (3.766 x 10−5 M), in contrast to a higher value for DBHCA (3.988 x 10−8 M). A comparison of the kinetic parameters [association constant (ka) and dissociation constant (kd) values] reveals that DBHCA can bind the target faster than DHNA, while the complex of DHNA with the target dissociates slower than that of DBHCA. Both DBHCA and DHNA can inhibit the growth of B. microti in vitro with half-maximal inhibitory concentration (IC50) values of 84.83 and 85.65 μM, respectively. Cytotoxicity tests in vitro further indicate that DBHCA and DHNA have selectivity indexes (SI) of 2.6 and 22.1 between B. microti and Vero cells, respectively. Although the two naphthalene-based compounds only display modest inhibitory activity against both rBmLDH and the growth of B. microti, the compound DHNA features high selectivity and could serve as a novel lead compound for designing LDH-specific antibabesial drug.

Abstract Background The spherical body, a membrane bound organelle localized in the apical organelle complex, is unique to Babesia and Theileria spp. The spherical body proteins (SBPs) secreted by spherical bodies include SBP1, SBP2, SBP3 and SBP4. Up to now, only SBP3 has been characterized in Babesia orientalis. Methods The BoSBP4 gene was amplified from cDNA and gDNA and cloned into the pGEX-6P-1 vector by homologous recombination, sequenced and analyzed by bioinformatics tools. The amino acid (aa) sequence of BoSBP4 was compared with that of Babesia bovis and Babesia bigemina as well as SBP3 of B. orientalis. The immunoreactivity was evaluated by incubating recombinant BoSBP4 (rBoSBP4) with the serum of B. orientalis-infected water buffalo. The native form of BoSBP4 was identified by incubating lysate of B. orientalis-infected water buffalo erythrocytes with the anti-rBoSBP4 mouse serum. The cellular localization of BoSBP4 was determined by indirect immunofluorescence assay. Results The full length of the BoSBP4 gene was estimated to be 945 bp without introns, encoding a 314 aa polypeptide with a predicted molecular weight of 37 kDa. The truncated recombinant protein was expressed from 70 to 945 bp as a GST fusion protein with a practical molecular weight of 70 kDa. BoSBP4 shared a 40% and 30% identity with B. bovis and B. bigemina, respectively. Furthermore, it was 31% identical to SBP3 of B. orientalis. BoSBP4 was identified in the lysate of B. orientalis-infected water buffalo erythrocytes with a molecular weight of 37 kDa, corresponding to the expected molecular mass of BoSBP4. The result of rBoSBP4 with positive serum revealed that BoSBP4 can elicit an immune response to B. orientalis-infected water buffalo. The cellular localization of BoSBP4 was detected to be adjacent to the merozoite nucleus in the intracellular phase, followed by the diffusion of the fluorescence of BoSBP4 into the cytoplasm of B. orientalis-infected erythrocytes as puncta-like specks and a gradual increase of the fluorescence. Conclusions In this study, SBP4 in B. orientalis was characterized for the first time. It may play a key role in interaction with the host cell by being secreted into the cytoplasm of the B. orientalis-infected erythrocytes to facilitate parasite growth and reproduction.

Nodes with high centrality in protein–protein interaction (PPI) networks are known to be essential in some organisms. Here, the authors in contrast find that in the interactome of A. thaliana central nodes are enriched in conditional and immune phenotypes and are preferred targets of pathogens.

Abstract Background The spherical body is a distinct organelle only existing in Babesia and Theileria. Spherical body proteins (SBPs) are secreted from spherical bodies and incorporated into the cytoplasm of infected erythrocytes during invasion and post-invasion stages. Four different SBP homologues (SBP1, SBP2, SBP3 and SBP4) have been identified in Babesia bovis and Babesia bigemina. So far, there has been no report available about the identification of SBPs in Babesia orientalis. Methods The SBP3-like in B. orientalis (BoSBP3-like) was cloned, sequenced, characterized and compared to the SBP3 sequences of B. bovis and B. bigemina by bioinformatics analyses. The BoSBP3-like gene was truncated into three fragments: BoSBP3-like-1 (915 bp), BoSBP3-like-2 (1311 bp) and BoSBP3-like-3 (1011 bp), which were amplified and cloned into the expression vector pET-28a and expressed as three truncated recombinant (His-fusion) proteins. The immunogenicity, native forms and localization of BoSBP3-like were identified by western blot and indirect immunofluorescence assay (IFA). Results The BoSBP3-like gene was intronless with an open reading frame (ORF) of 3237 bp, encoded a 1079 amino acid polypeptide with a predicted size of 135 kDa, and contained a cysteine-rich region, three dispersing FAINT domains and a signal peptide (1–16 aa) at the N-terminus. The amino acid sequence of BoSBP3-like was 61.6 and 35.0% identical to that of B. bovis and B. bigemina, respectively. BoSBP3-like was identified as 135 kDa in the parasite lysate by rabbit antiserum against the truncated recombinant BoSBP3-like-1 (rBoSBP3-like-1). Three specific bands corresponding to rBoSBP3-like-1 (1–305 aa, 43 kDa), rBoSBP3-like-2 (306–742 aa, 58 kDa) and rBoSBP3-like-3 (743–1079 aa, 52 kDa) were detected by reaction with serum from B. orientalis-infected buffalo. The BoSBP3-like was not only localized in the spherical body of B. orientalis but also in the cytoplasm of infected erythrocytes of buffalo as puncta-like protein specks at both single and paired parasite development stages. Conclusions Through secretion into the cytoplasm of infected erythrocytes, BoSBP3-like may play a significant role in adaptation, interaction, and modification related to the host environment to benefit the growth and survival of Babesia. BoSBP3-like could react with the serum from B. orientalis-infected buffalo, but not healthy buffalo, implicating that BoSBP3-like is highly antigenic and may serve as a candidate diagnostic antigen for the detection of B. orientalis.

Abstract The universal nature of networks’ structural and physical properties across diverse systems offers a better prospect to elucidate the interplay between a system and its environment. In the last decade, several large-scale transcriptome and interactome studies were conducted to understand the complex and dynamic nature of interactions between Arabidopsis and its bacterial pathogen, Pseudomonas syringae pv. tomato DC3000. We took advantage of these publicly available datasets and performed “-omics”-based integrative, and network topology analyses to decipher the transcriptional and protein-protein interaction activities of effector targets. We demonstrated that effector targets exhibit shorter distance to differentially expressed genes (DEGs) and possess increased information centrality. Intriguingly, effector targets are differentially expressed in a sequential manner and make for 1% of the total DEGs at any time point of infection with virulent or defense-inducing DC3000 strains. We revealed that DC3000 significantly alters the expression levels of 71% effector targets and their downstream physical interacting proteins in Arabidopsis interactome. Our integrative “-omics”-–based analyses identified dynamic complexes associated with MTI and disease susceptibility. Finally, we discovered five novel plant defense players using a systems biology-fueled top-to-bottom approach and demonstrated immune-related functions for them, further validating the power and resolution of our network analyses.

SCOPE: Circadian rhythms allow organisms to anticipate and exploit daily environmental changes, and food components can adjust internal rhythms in peripheral tissues or even suprachiasmatic nucleus. Dysregulation of circadian rhythms is associated with many pathological conditions. The aim of the current study is to determine potential effects of tea polyphenols on modulating circadian rhythms regarding ameliorating diabetes, metabolic syndrome, and memory disorders. Results: Results demonstrated that tea polyphenols can reverse the relatively shallow daily oscillations of circadian clock genes transcription and protein expression in both liver and hypothalamus triggered by constant darkness. These compounds also alleviate insulin resistance and disorder of glucose/lipid metabolism in constant darkness-treated mice. Concomitantly, tea polyphenols prevented constant darkness-induced memory impairment by inhibiting neuron damage and elevating the expression of neurotropic factor related genes. Conclusion: Tea polyphenols supplementation might serve as future strategies to combat obesity, metabolic syndrome and cognitive disorder induced by circadian dysfunction.

A number of studies have demonstrated that Babesia orientalis (B. orientalis) can only infect water buffalo (Bubalus bubalis) and not dairy cattle (Bos taurus) or beef cattle (Bos taurus), even though all three belong to the tribe Bovini and have close evolutionary relationships. In addition, Babesia species are intracellular protozoans that obligately parasitize in erythrocytes. This may indicate that the infection specificity is due to differences in erythrocyte proteins. Totals of 491, 1,143, and 1,145 proteins were identified from water buffalo, beef cattle, and dairy cattle, respectively, by searching the Uniprot and NCBI databases. The number of proteins identified for water buffalo was far lower than for beef cattle and dairy cattle, particularly in the range from 15 to 25 kDa. Remarkably, 290 identified proteins were unique to water buffalo, of which putative gamma-globin and putative epsilon-globin had a significant possibility of being relevant to the survival of B. orientalis only in water buffalo. A total of 2,222 proteins were annotated in terms of molecular function, biological process, and cellular component according to GO annotation. The number of proteins of water buffalo in oxygen binding was far higher than for beef cattle and dairy cattle. This is the first time that the protein profiles of water buffalo, beef cattle, and dairy cattle have been comparatively analyzed. The uniquely expressed proteins in water buffalo obtained in this study may provide new insights into the mechanism of B. orientalis infection exclusivity in water buffalo and may be a benefit for the development of strategies against B. orientalis.

In plants, programmed cell death (PCD) has diverse, essential roles in vegetative and reproductive development, and in the responses to abiotic and biotic stresses. Despite the rapid progress in understanding the occurrence and functions of the diverse forms of PCD in plants, the signaling components and molecular mechanisms underlying the core PCD machinery remain a mystery. The roles of BAK1 (BRASSINOSTEROID INSENSITIVE 1-associated receptor kinase 1), an essential co-receptor of multiple receptor complexes, in the regulation of immunity and development- and defense-related PCD have been well characterized. However, the ways in which BAK1 functions in mediating PCD need to be further explored. In this review, different forms of PCD in both plants and mammals are discussed. Moreover, we mainly summarize recent advances in elucidating the functions and possible mechanisms of BAK1 in controlling diverse forms of PCD. We also highlight the involvement of post-translational modifications (PTMs) of multiple signaling component proteins in BAK1-mediated PCD.

Toxoplasma gondii belongs to the Apicomplexan protozoa—an obligate intracellular parasite—causing toxoplasmosis that has a worldwide distribution and is very harmful to both human health and the livestock industry. However, the information on toxoplasmosis in the Qinghai-Tibetan Plateau Area (QTPA) and the seroprevalence of T. gondii in the food-borne animals in that area has been limited. Therefore, this study focused to T. gondii and toxoplasmosis to perform an indirect ELISA test based on recombinant TgSAG2 protein to establish a comprehensive record of the seroprevalence of T. gondii infections in a wide range of animals, including Tibetan sheep (Ovis aries), yaks (Bos grunniens), cows, chicken, pigs, and horses, in the QTPA. Overall, the seropositive rates of the specific-T. gondii IgG and IgM antibodies in all investigated animals were 44.1% (1179/2673) and 18.0% (469/2612), respectively. The 14.9% (389/2612) sera were determined to be both IgG and IgM positive samples, 30.2% (789/2673) were single-IgG seropositive, and a total of 80 in 2612 animals (3.0%) were single-IgM seropositive. Moreover, for the animal species, the pig was the most prevalent animal (90.2%, 304/337) for IgG positivity, followed by Tibetan sheep (50.7%, 460/907), chickens (45.8%, 229/500), yaks (21.1%, 140/663), cows (18.5%, 38/205) and horses (13.1%, 8/61), respectively. For the IgM antibody positivity, the pig was also the most prevalent animal (41.8%, 141/337), followed by Tibetan sheep (21.2%, 191/907), cows (15.1%, 31/205), chickens (12.4%, 62/500) and yaks (6.6%, 44/663), respectively. The significant differences in the prevalent distribution of T. gondii were found in the different altitudes. In conclusion, this study found the high seroprevalence for T. gondii infections among these animal species in the QTPA, and provides new data to facilitate further research for development of control measures against T. gondii infections in the surveyed locations.

Gibberella stalk rot (GSR) by Fusarium graminearum causes significant losses of maize production worldwide. Jasmonates (JAs) have been broadly known in regulating defense against pathogens through the homeostasis of active JAs and COI-JAZ-MYC function module. However, the functions of different molecular species of JAs and COI-JAZ-MYC module in maize interactions with Fusarium graminearum and regulation of diverse metabolites remain unknown. In this study, we found that exogenous application of MeJA strongly enhanced resistance to GSR. RNA-seq analysis showed that MeJA activated multiple genes in JA pathways, which prompted us to perform a genome-wide screening of key JA signaling components in maize. Yeast Two-Hybrid, Split-Luciferase, and Pull-down assays revealed that the JA functional and structural mimic coronatine (COR) functions as an essential ligand to trigger the interaction between ZmCOIa and ZmJAZ15. By deploying CRISPR-cas9 knockout and Mutator insertional mutants, we demonstrated that coi1a mutant is more resistant, whereas jaz15 mutant is more susceptible to GSR. Moreover, JA-deficient opr7-5opr8-2 mutant displayed enhanced resistance to GSR compared to wild type. Together, these results provide strong evidence that ZmJAZ15 plays a pivotal role, whereas ZmCOIa and endogenous JA itself might function as susceptibility factors, in maize immunity to GSR.

Anosovite crystalline is an ideal mineral for flotation from the Ti-bearing blast furnace (TBBF) slag. Ti3O5 crystal and Al2TiO5 crystal are two kinds of anosovites, and the Al element significantly affects the electronic structure and flotation performance of anosovite. The floatability of Ti3O5 and Al2TiO5 crystals were studied by Mulliken populations, energy bands, and density of states (DOS). In addition, the flotation experiment of the two kinds of anosovite crystals (Ti3O5 and Al2TiO5) was conducted and proved that the density functional theory (DFT) calculation results were accurate. Compared with Ti3O5 crystal, the Fermi energy level of Al2TiO5 crystal shifts around 2 eV in a negative direction by DOS analysis, which is beneficial to flotation. And Al2TiO5 crystal possesses a larger value of bond population, which is 0.41, for Ti-O bonds than Ti3O5 crystal and the bond length of Ti-O in Al2TiO5 crystal is shorter, therefore Al2TiO5 crystal shows a stronger covalency. The changes of the Fermi energy level and the covalency bonds in Al2TiO5 crystal both demonstrated that doping the Al component into the Ti3O5 crystal was beneficial to improve the flotation effect. Moreover, the Al2TiO5 crystal had a higher flotation efficiency compared to the Ti3O5 crystal when the dosages of salicylhydroxamic acid (SHA) and sodium oleate were the same. Therefore, both DFT calculation and experiment show that the flotation effect of the Al2TiO5 crystal is better than that of the Ti3O5 crystal.

Corn stalk rot caused by Fusarium spp., a genus of soil-borne fungal pathogens, has become a major concern of maize production. This disease normally causes significant reduction of maize yield and quality worldwide. The field assay for identifying stalk rot resistance using adult plants is largely relying on large population, yet time-consuming, labor costs, and often influenced by environmental conditions. Therefore, a rapid and reliable assay for investigating maize stalk rot caused by Fusarium spp. is required for screening the resistant lines and functional study of maize resistance to this pathogen. We have developed a seedling assay to rapidly screen the resistant lines using 12-day to 2-week-old seedlings. The entire assay can be completed within approximately 16-18 days post seed germination, with inexpensive labor cost and high repeatability. This simple, rapid and reliable assay can be widely used for identifying the maize resistance to stalk rot caused by Fusarium spp. and other similar fungal pathogens.

Babesiosis caused by Babesia orientalis is one of the most prevalent infections of water buffalo transmitted by Rhipicephalus haemaphysaloides causing a parasitic and hemolytic disease. The organelles proteins localized in apical membrane especially rhoptries neck and microneme protein form a complex called moving junction important during invasion process of parasites belonging to apicomplexan group, including Babesia species. A truncated fragment coding a 936 bps fragment was cloned in pMD-19T and subcloned into pET32 (a)+ expression vector, expressed in E. coli BL21. Purified recombinant BoRON2 was used to produce polyclonal antibody against BoRON2. Here, we identified the full sequence of gene encoding the rhoptry neck 2 protein that we named BoRON2 which is 4035 bp in full-length open reading frame without introns, encoding a polypeptide of 1345 amino acids. Western blot of rBoRON2 probed with buffalo positive serum analysis revealed a band of around 150 kDa in parasite lysates, suggesting an active involvement during invasion process. These findings most likely are constructive in perspective of ongoing research focused particularly on water buffalo babesiosis prevention and therapeutics and globally provide new information for genes comparative analysis.

Some coronaviruses (CoVs) have an extra furin cleavage site (RRKR/S, furin-S2′ site) upstream of the fusion peptide in the spike protein, which plays roles in virion adsorption and fusion. Mutation of the S2′ site of QX genotype (QX-type) infectious bronchitis virus (IBV) spike protein (S) in a recombinant virus background results in higher pathogenicity, pronounced neural symptoms and neurotropism when compared with conditions in wild-type IBV (WT-IBV) infected chickens. In this study, we present evidence suggesting that recombinant IBV with a mutant S2′ site (furin-S2′ site) leads to higher mortality. Infection with mutant IBV induces severe encephalitis and breaks the blood−brain barrier. The results of a neutralization test and immunoprotection experiment show that an original serum and vaccine can still provide effective protection in vivo and in vitro. This is the first demonstration of IBV-induced neural symptoms in chickens with encephalitis and the furin-S2′ site as a determinant of neurotropism.