Comprehensive genomic and metabolomic analysis revealed the physiological characteristics and pickle like odor compounds metabolic pathways of Bacillus amyloliquefaciens ZZ7 isolated from fermented grains of Maotai-flavor baijiu.

Pickle like odor (PLO) is one of the main defective flavors of Maotai flavor baijiu (MFB). Understanding and controlling the PLO compounds producing strains not only solves the problem of PLO from the source, but also ensures the high-quality production of MFB. However, the relevant research on PLO compounds producing strains has not been reported in MFB. In this study, we identified a Bacillus amyloliquefaciens ZZ7 with high yield of PLO compounds in the fermented grains of MFB, and measured its physiological characteristics. It produces 627 volatile compounds and 1,507 non-volatile compounds. There are 7 volatile sulfur compounds that cause the PLO, the content of dimethyl disulfide, dimethyl trisulfide, and dimethyl sulfur is relatively high, accounting for 89.43% of the total volatile sulfur compounds. The genome size of B. amyloliquefaciens ZZ7 is 3,902,720 bp with a GC content of 46.09%, and a total of 3,948 protein coding genes were predicted. Moreover, the functional annotation of coding genes and an assessment of the metabolic pathways were performed by genome annotation, showing it has strong ability to transport and metabolize amino acids and carbohydrates. Comprehensive genomic and metabolomic analysis, the metabolic pathway of PLO compounds of B. amyloliquefaciens ZZ7 was revealed, which mainly involves 12 enzymes including sulfate adenylyltransferase, cysteine synthase, cystathionine γ-synthase, etc. This work provides biological information support at both genetic and metabolic levels for the mechanism of B. amyloliquefaciens ZZ7 to synthesize PLO compounds, and provides a direction for the subsequent genetic modification of ZZ7 to solve PLO from the source in the MFB.

Comparison of Synthetic Methods and Identification of Several Artificial Antigens of Deoxynivalenol.

The purpose of this experiment was to study the design and modification of hapten molecules and artificial antigen molecules of deoxynivalenol (DON), and to compare the preparation and identification methods of four artificial antigens. According to the characteristics of the molecular structure of DON, four artificial antigen coupling methods were designed-namely, N,N'-carbonyldiimidazole (CDI), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC), isobutyl chloroformate (IBCF), and N-hydroxysuccinimide (NHS)-to prepare artificial antigens and detection antigens. Through ultraviolet (UV), infrared (IR), and SDS-polyacrylamide gel electrophoresis (SDS-PAGE), along with other physical and chemical identification methods and animal immunisation, the best artificial antigen coupling method was screened. The results showed that the CDI method achieved the best effect among the synthesis methods. The titre of anti-DON polyclonal antibody (pAb) produced by animal immunisation reached 1: (6.4 × 103). The half inhibitory concentration (IC50) was 47.75 ng/mL, the cross-reaction rate with 3-acetyldeoxynivalenol (3-AcDON) was slightly higher at 35.3%, and there was no cross-reaction with other compounds; therefore, four artificial antigens were successfully prepared by using the molecular structure of DON. Through identification, the CDI method was screened as the best artificial antigen synthesis method, with the highest DON pAb titre, the best sensitivity, and the strongest specificity. This will lay a solid antigenic foundation for the preparation of better anti-DON monoclonal antibodies (mAbs) in the future.

Electrical Broth Micro-Dilution for Rapid Antibiotic Resistance Testing.

Rapid tests to assess the susceptibility of bacteria to antibiotics are required to inform antibiotic stewardship. We have developed a novel test, which measures changes in the impedance of a 100 nanoliter volume of bacterial suspension to determine an "electrical" minimum inhibitory concentration (eMIC). Two representative strains of Klebsiella pneumoniae, Acinetobacter baumannii, Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus were tested against a panel of frontline antibiotics with different modes of action (ciprofloxacin, doxycycline, colistin and imipenem, gentamicin, and ceftazidime). The eMIC measured at 1 h correlated strongly with a standard 24 h microbroth dilution MIC for all combinations of antibiotics and bacteria, allowing strains to be correctly assigned as sensitive or resistant measured in a fraction of the time.

Inhibiting the Dissolution of Lithium Polyphosphides and Enhancing the Reaction Kinetics of a Phosphorus Anode via Screening Functional Additives.

A high-capacity, low-cost phosphorus anode is considered as one of the most promising candidates for next-generation Li-ion batteries. Nevertheless, the dissolution/shuttle effect of lithium polyphosphides and sluggish electrochemical conversion hinder the practical application of a phosphorus anode, similar to the problems of a sulfur cathode. Although the reported functional additives with physical obstruction and chemical adsorption have been successful in improving the performance of a sulfur cathode, they can not be directly applied to phosphorus due to their deterioration and failure in low voltage. To solve the above problems, we made a systematic investigation to rationally select the functional additives (Li2O, Li2S, and LiF) and effectively guide the experiment. These functional additives possess synergetic effects, including the adsorption of soluble lithium polyphosphides and the catalytic conversion of phosphorus species. The design of these functional additives provides a guiding and screening principle for inhibiting the dissolution of polyphosphides and improving the reaction kinetics of a phosphorus anode.

Plant Disease Resistance-Related Signaling Pathways: Recent Progress and Future Prospects.

Plant-pathogen interactions induce a signal transmission series that stimulates the plant's host defense system against pathogens and this, in turn, leads to disease resistance responses. Plant innate immunity mainly includes two lines of the defense system, called pathogen-associated molecular pattern-triggered immunity (PTI) and effector-triggered immunity (ETI). There is extensive signal exchange and recognition in the process of triggering the plant immune signaling network. Plant messenger signaling molecules, such as calcium ions, reactive oxygen species, and nitric oxide, and plant hormone signaling molecules, such as salicylic acid, jasmonic acid, and ethylene, play key roles in inducing plant defense responses. In addition, heterotrimeric G proteins, the mitogen-activated protein kinase cascade, and non-coding RNAs (ncRNAs) play important roles in regulating disease resistance and the defense signal transduction network. This paper summarizes the status and progress in plant disease resistance and disease resistance signal transduction pathway research in recent years; discusses the complexities of, and interactions among, defense signal pathways; and forecasts future research prospects to provide new ideas for the prevention and control of plant diseases.

Construction of Recombinant Rabies Virus Vectors Expressing H or F Protein of Peste des Petits Ruminants Virus.

Peste des petits ruminants (PPR) is one of the most contagious and fatal diseases of small ruminants in the world and is classified as a category A epidemic disease. It is the target of a global eradication campaign led by the Office International des Epizooties (OIE) and Food and Agriculture Organization of the United Nations (FAO). The PPR live attenuated vaccine is currently the most widely used and approved vaccine, but the use of this vaccine interferes with the serological testing of the PPR elimination program, and there is a potential safety risk. Viral vector vaccines are one of the most promising methods to solve this dilemma. In this study, the full-length infectious clone plasmid of rabies virus (RABV), pD-SRV9-PM-LASV, was used as the backbone, and the envelope glycoprotein H (hemagglutinin protein) or F (fusion protein) gene of PPRV was inserted into the backbone plasmid to construct the infectious clones pD-SRV9-PM-PPRV-H and pD-SRV9-PM-PPRV-F, which express the PPRV H and PPRV F genes, respectively. The correct construction of these infectious clones was verified after sequencing and double digestion. The infectious clones were transfected with a helper plasmid into BSR/T7 cells, and recombinant viruses were successfully rescued by direct immunofluorescence, indirect immunofluorescence, Western blotting, and transmission electron microscopy and named rSRV9-H and rSRV9-F. The results of growth kinetics studies indicated that the inserted gene did not affect virus proliferation. Stability studies revealed that the inserted target gene was stably expressed in recombinant RABV for at least 15 generations. In this study, the recombinant viruses rSRV9-H and rSRV9-F were successfully rescued. The constructed viruses had good proliferative activity and stability and provided potential bivalent inactivated vaccine candidate strains for the prevention of PPR and livestock rabies.

Characteristics of Chimeric West Nile Virus Based on the Japanese Encephalitis Virus SA14-14-2 Backbone.

West Nile virus disease (WND) is an arthropod-borne zoonosis responsible for nonspecific fever or severe encephalitis. The pathogen is West Nile virus belonging to the genus Flavivirus, family Flaviviridae. Every year, thousands of cases were reported, which poses significant public health risk. Here, we constructed a West Nile virus chimera, ChiVax-WN01, by replacing the prMΔE gene of JEV SA14-14-2 with that of the West Nile virus NY99. The ChiVax-WN01 chimera showed clear, different characters compared with that of JEV SA14-14-2 and WNV NY99 strain. An animal study indicated that the ChiVax-WN01 chimera presented moderate safety and immunogenicity for 4-week female BALB/c mice.

Immunogenicity Assessment of Rift Valley Fever Virus Virus-Like Particles in BALB/c Mice.

Rift Valley fever (RVF) is an acute, febrile zoonotic disease that is caused by the RVF virus (RVFV) and is spread by arthropod vectors. Virus-like particle (VLP) vaccines, which have the advantages of strong immunogenicity and safety, play an important role in the prevention of this disease. VLPs for RVFV were successfully prepared by our research group using a baculovirus-insect cell expression system. To study the immunogenicity of these RVFV VLPs, a correct 3rd or 4th generation recombinant baculovirus, rBac-N-G, was identified and used to infect Sf9 cells, which were cultured in suspension at a large scale. Subsequently, cell debris was removed by centrifugation, and the VLPs were concentrated by ultracentrifugation and purified using a sucrose gradient, after which they were used to immunize BALB/c mice by intramuscular injection. The results showed that the RVFV VLPs prepared by our research group could effectively induce mice to produce RVFV neutralizing antibodies and that the prepared VLPs could stimulate mouse spleen cells to produce high levels of the cytokines IL-4 and IFN-γ. Moreover, the proportion of lymphocytes producing IL-4 and IFN-γ in the spleen of mice immunized with RVFV VLPs was significantly increased. Therefore, the RVFV VLPs prepared in this study had strong immunogenicity and could effectively activate humoral and cellular immunity in mice. This study lays a solid foundation for the development of RVFV VLP vaccine candidates and promotes the healthy development of animal husbandry and human public health.

Development of a rapid phenotypic test on a microfluidic device for carbapenemase detection using the chromogenic compound nitrocefin.

Routine identification of carbapenemase-producing bacterial isolates is a lengthy process often taking up to 72 h to generate results with standard culture-based tests. Here we describe a rapid test based on the hydrolysis of nitrocefin to identify isolates producing ß-lactamase enzymes. A cocktail of inhibitors has been optimized in the reaction mix to provide specificity for carbapenemase enzymes. The developed assay has also been translated to a microfluidic platform with an optical readout (optofluidic chip). The chip has a long absorbance path (25 mm) to provide high sensitivity. A sample-to-answer has been achieved in under 30 min on these chips using colonies from culture plates. The test on this platform has the potential to provide a rapid indicative (presumptive positive) test for carbapenemase producers direct from bacteria isolated from patient samples, to rapidly trigger infection control measures and identify samples that should be prioritized for more specialized carbapenemase diagnostic assays.

Development of a Direct Competitive ELISA Kit for Detecting Deoxynivalenol Contamination in Wheat.

This study was conducted to develop a self-assembled direct competitive enzyme-linked immunosorbent assay (dcELISA) kit for the detection of deoxynivalenol (DON) in food and feed grains. Based on the preparation of anti-DON monoclonal antibodies, we established a standard curve with dcELISA and optimized the detection conditions. The performance of the kit was evaluated by comparison with high-performance liquid chromatography (HPLC). The minimum detection limit of DON with the kit was 0.62 ng/mL, the linear range was from 1.0 to 113.24 ng/mL and the half-maximal inhibition concentration (IC50) was 6.61 ng/mL in the working buffer; there was a limit of detection (LOD) of 62 ng/g, and the detection range was from 100 to 11324 ng/g in authentic agricultural samples. We examined four samples of wheat bran, wheat flour, corn flour and corn for DON recovery. The average recovery was in the range of 77.1% to 107.0%, and the relative standard deviation (RSD) ranged from 4.2% to 11.9%. In addition, the kit has the advantages of high specificity, good stability, a long effective life and negligible sample matrix interference. Finally, wheat samples from farms in the six provinces of Henan, Anhui, Hebei, Shandong, Jiangsu and Gansu in China were analyzed by the kit. A total of 30 samples were randomly checked (five samples in each province), and the results were in good agreement with the standardized HPLC method. These tests showed that the dcELISA kit had good performance and met relevant technical requirements, and it had the characteristics of accuracy, reliability, convenience and high-throughput screening for DON detection. Therefore, the developed kit is suitable for rapid screening of DON in marketed products.

Equine immunoglobulin F(ab')2 fragments protect mice from Rift Valley fever virus infection.

BACKGROUND: Rift Valley fever virus (RVFV) is an emerging arbovirus in Africa and the Arabian Peninsula, in which infection with RVFV poses a serious threat to humans and livestock globally. Approved treatments for RVFV infection, especially for use in humans, have not yet been developed. There is an urgent need for effective drugs to prevent RVFV disease. METHODS: In previous study, we developed RVFV virus like particles (VLPs) expressing the surface glycoproteins Gn and Gc. The morphology was shown to be similar to live RVFV under electron microscopy. In this study, we immunized horses with RVFV VLPs, prepared the immunoglobulin F(ab')2 fragments, and characterized its in vitro neutralization and in vivo efficacy in mice. RESULTS: F(ab')2 was found to potently neutralize RVFV in VeroE6 cells, and passive transfer of immunoglobulin F(ab')2 fragments resulting in reduced mortality in RVFV infected mice. CONCLUSION: Our results show that passive immunotherapy with equine immunoglobulin F(ab')2 fragments is a promising strategy to treat RVFV infections.

Packaging of Rift Valley fever virus pseudoviruses and establishment of a neutralization assay method.

Rift Valley fever (RVF) is an acute, febrile zoonotic disease that is caused by the RVF virus (RVFV). RVF is mainly prevalent on the Arabian Peninsula, the African continent, and several islands in the Indian Ocean near southeast Africa. RVFV has been classified by the World Organisation for Animal Health (OIE) as a category A pathogen. To avoid biological safety concerns associated with use of the pathogen in RVFV neutralization assays, the present study investigated and established an RVFV pseudovirus-based neutralization assay. This study used the human immunodeficiency virus (HIV) lentiviral packaging system and RVFV structural proteins to successfully construct RVFV pseudoviruses. Electron microscopy observation and western blotting indicated that the size, structure, and shape of the packaged pseudoviruses were notably similar to those of HIV lentiviral vectors. Infection inhibition assay results showed that an antibody against RVFV inhibited the infective ability of the RVFV pseudoviruses, and an antibody neutralization assay for RVFV detection was then established. This study has successfully established a neutralization assay based on RVFV pseudoviruses and demonstrated that this method can be used to effectively evaluate antibody neutralization.

Electrically tunable whispering gallery mode microresonator based on a grapefruit-microstructured optical fiber infiltrated with nematic liquid crystals.

An electrically tunable whispering gallery mode (WGM) microresonator based on an HF-etched microstructured optical fiber (MOF) infiltrated with nematic liquid crystals (NLCs) is proposed and experimentally demonstrated. Experimental results indicate that as the peak-to-peak voltage of the applied AC electric field increases from 160 to 220 V, WGM resonance peaks gradually move toward a shorter wavelength region by 0.527 nm with a wavelength sensitivity up to 0.01 nm/V for a TM1691 mode, and the Q-factor for each WGM resonance peak rapidly decreases with the increment of applied electric voltage. The proposed electrically controlled WGM tuning scheme shows a linear resonance wavelength shift with good spectral reversibility, which makes it a promising candidate to serve as an integrated functional photonic device in practical use and in related fundamental scientific studies.

Treatment with hyperimmune equine immunoglobulin or immunoglobulin fragments completely protects rodents from Ebola virus infection.

Recent successes with monoclonal antibody cocktails ZMapp(TM) and MIL77 against Ebola virus (EBOV) infections have reignited interest in antibody-based therapeutics. Since the production process for monoclonal antibodies can be prolonged and costly, alternative treatments should be investigated. We produced purified equine antisera from horses hyperimmunized with EBOV virus-like particles, and tested the post-exposure efficacy of the antisera in a mouse model of infection. BALB/c mice were given up to 2 mg of purified equine antisera per animal, at 30 minutes, 1 or 2 days post-infection (dpi), in which all animals survived. To decrease the possibility of serum sickness, the equine antisera was digested with pepsin to generate F(ab')2 fragments, with in vitro neutralizing activity comparable to whole immunoglobulin. Full protection was achieved with when treatment was initiated at 1 dpi, but the suboptimal protection observed with the 30 minute and 2 dpi groups demonstrate that in addition to virus neutralization, other Fc-dependent antibody mechanisms may also contribute to survival. Guinea pigs given 20 mg of antisera or F(ab')2 at or starting at 1 or 2 dpi were also fully protected from EBOV infection. These results justify future efficacy studies for purified equine products in NHPs.

Effects of longitudinal body mass index variability on microvasculature over 5 years in adult Chinese.

OBJECTIVE: To explore the associations of 5-year trend and fluctuation in body mass index (BMI) with retinal vascular caliber in a middle-aged and elderly Chinese population. METHODS: Participants age ≥40 years were recruited in a prospective study. Baseline BMI data were collected in 2008, and the participants were re-examined annually until 2012. Retinal vascular caliber was measured from fundus photographs collected in 2012. BMI trend was calculated as the slope of BMI against the time of examinations. BMI fluctuation was defined as the root mean square error around the regression line of BMI over time (BMI RMSE) and the coefficient of variation of BMI (BMI CV). RESULTS: Rising BMI trend was associated with narrower retinal arteriolar and wider venular calibers in the overall subjects, especially among persons with overweight and obesity (BMI ≥ 25 kg/m(2) , P = 0.004 and 0.033, respectively). Rising BMI trend was also significantly associated with narrower retinal arteriole even in nonobese individuals with BMI < 25 kg/m(2) (P = 0.017) when eliminating the effects of hypertension and diabetes. Neither BMI RMSE nor BMI CV was statistically associated with retinal vascular caliber (all P > 0.05). CONCLUSIONS: Annual rising trending BMI was associated with retinal microvascular alteration. The results suggest that weight gain probably increases the risk of cardiovascular diseases among middle-aged and elderly people.

Laser-tuned whispering gallery modes in a solid-core microstructured optical fibre integrated with magnetic fluids.

A laser-assisted tuning method of whispering gallery modes (WGMs) in a cylindrical microresonator based on magnetic-fluids-infiltrated microstructured optical fibres (MFIMOFs, where MF and MOF respectively refer to magnetic fluid and microstructured optical fibre) is proposed, experimentally demonstrated and theoretically analysed in detail. The MFIMOF is prepared by infiltrating the air-hole array of the MOF using capillary action effect. A fibre-coupling system is set up for the proposed MFIMOF-based microresonator to acquire an extinction ratio up to 25 dB and a Q-factor as large as 4.0 × 10(4). For the MF-infiltrated MOF, the light propagating in the fibre core region would rapidly spread out and would be absorbed by the MF-rod array cladding to induce significant thermal effect. This has been exploited to achieve a WGM resonance wavelength sensitivity of 0.034 nm/mW, which is ~20 times higher than it counterpart without MF infiltration. The wavelength response of the resonance dips exhibit linear power dependence, and owing to such desirable merits as ease of fabrication, high sensitivity and laser-assisted tunability, the proposed optical tuning approach of WGMs in the MFIMOF would find promising applications in the areas of optical filtering, sensing, and signal processing, as well as future all-optical networking systems.

Intraocular pressure, central corneal thickness, and glaucoma in chinese adults: the liwan eye study.

PURPOSE: To describe the distribution of central corneal thickness (CCT), intraocular pressure (IOP), and their determinants and association with glaucoma in Chinese adults. DESIGN: Population-based cross-sectional study. METHODS: Chinese adults aged 50 years and older were identified using cluster random sampling in Liwan District, Guangzhou. CCT (both optical [OCCT] and ultrasound [UCCT]), intraocular pressure (by Tonopen, IOP), refractive error (by autorefractor, RE), radius of corneal curvature (RCC), axial length (AL), and body mass index (BMI) were measured, and history of hypertension and diabetes (DM) was collected by questionnaire. Right eye data were analyzed. RESULTS: The mean values of OCCT, UCCT, and IOP were 512 ± 29.0 µm, 542 ± 31.4 µm, and 15.2 ± 3.1 mm Hg, respectively. In multiple regression models, CCT declined with age (P < .001) and increased with greater RCC (P < .001) and DM (P = .037). IOP was positively associated with greater CCT (P < .001), BMI (P < .001), and hypertension (P < .001). All 25 persons with open-angle glaucoma had IOP <21 mm Hg. CCT did not differ significantly between persons with and without open- or closed-angle glaucoma. Among 65 persons with ocular hypertension (IOP >97.5th percentile), CCT (555 ± 29 µm) was significantly (P = .01) higher than for normal persons. CONCLUSIONS: The distributions of CCT and IOP in this study are similar to that for other Chinese populations, though IOP was lower than for European populations, possibly due to lower BMI and blood pressure. Glaucoma with IOP <21 mm Hg is common in this population. We found no association between glaucoma and CCT, though power (0.3) for this analysis was low.

Refractive error and biometry in older Chinese adults: the Liwan eye study.

PURPOSE: To assess the prevalence of refractive error and describe the distribution of ocular biometry and its association with refraction in adult Chinese. METHODS: Random clustering sampling was used to identify adults aged > or = 50 years in Liwan District, Guangzhou. Refraction was determined by subjective refraction that achieved the best corrected vision based on monocular measurement. Ocular biometry was measured by A-mode ultrasound using a handheld applanation probe. RESULTS: Among 1405 participants in the study, data from 1269 phakic right eyes were available for analysis. The prevalence of myopia (SE < -0.5 D), hyperopia (SE > +0.5 D), and astigmatism (cylinder > 0.75 D) was 32.3% (95% confidence interval [CI], 27.8%-34.6%), 40.0% (95% CI, 37.3%-42.7%), and 48.3% (95% CI, 45.6%-51.1%), respectively. The spherical equivalent tended to become hyperopic at 60 years and shifted toward myopia at 75 years. Axial length did not change with age but was consistently shorter in women. Lens thickness increased with age and tended to be greater in women. CONCLUSIONS: The prevalence of myopia and biometric distribution in this urban Chinese cohort are similar to those observed in Singaporean Chinese but greater than in Mongolians and Europeans. Further studies are needed to clarify the role of environmental factors in the myopia rates.