Assessing heavy metal contamination in Amomum villosum Lour. fruits from plantations in Southern China: Soil-fungi-plant interactions.

Amomum villosum Lour. fruit is a common healthy food widely cultivated in southern China. Heavy metal contamination of farmland soils has becomes a serious environmental concern in China. Heavy metals in soil can be introduced into the food chain and pose health risks to humans. However, microbial communities may play beneficial roles in plants grown in metal-polluted soils. This study aimed to assess the potential health risks of heavy metals in soils and A. villosum fruits from different production areas and to explore the soil-microbe-plant regulation pattern for heavy metals in A. villosum fruits. Soil and A. villosum fruit samples were collected from nine planting fields in four provinces of southern China. The results showed that soils from seven areas were polluted with heavy metals to different degrees. Cr and Mn were the most serious contaminating elements. However, the accumulation of heavy metals in A. villosum fruit was negligible with no expected human health risks. Partial least squares path analysis of structural equation modeling showed that the accumulation of heavy metals in A. villosum fruits was influenced by multiple factors. More importantly, the PLS-SEM revealed that the heavy metal content in A. villosum fruits was indirectly affected by soil heavy metals through the regulation of the microbial community. Furthermore, some fungal phyla (e.g., Ascomycota and Chytridiomycota) and genera (e.g., Mucor) were related to the heavy metal content in the soil and in A. villosum fruits. The results of this study verified that soil fungal community play an important role in the accumulation of heavy metals in A. villosum fruits. Using fungi provides a potential biological strategy for reducing the health risk posed by heavy metals in food.

Neural representation of phonological information during Chinese character reading.

Previous studies have revealed that phonological processing of Chinese characters elicited activation in the left prefrontal cortex, bilateral parietal cortex, and occipitotemporal regions. However, it is controversial what role the left middle frontal gyrus plays in Chinese character reading, and whether the core regions (e.g., the left superior temporal gyrus and supramarginal gyrus) for phonological processing of alphabetic languages are also involved in Chinese character reading. To address these questions, the present study used both univariate and multivariate analysis (i.e., representational similarity analysis, RSA) to explore neural representations of phonological information during Chinese character reading. Participants were scanned while performing a reading aloud task. Univariate activation analysis revealed a widely distributed network for word reading, including the bilateral inferior frontal gyrus, middle frontal gyrus, lateral temporal cortex, and occipitotemporal cortex. More importantly, RSA showed that the left prefrontal (i.e., the left middle frontal gyrus and left inferior frontal gyrus) and bilateral occipitotemporal areas (i.e., the left inferior and middle temporal gyrus and bilateral fusiform gyrus) represented phonological information of Chinese characters. These results confirmed the importance of the left middle frontal gyrus and regions in ventral pathway in representing phonological information of Chinese characters.

Language distance in orthographic transparency affects cross-language pattern similarity between native and non-native languages.

How native and non-native languages are represented in the brain is one of the most important questions in neurolinguistics. Much research has found that the similarity in neural activity of native and non-native languages are influenced by factors such as age of acquisition, language proficiency, and language exposure in the non-native language. Nevertheless, it is still unclear how the similarity between native and non-native languages in orthographic transparency, a key factor that affects the cognitive and neural mechanisms of phonological access, modulates the cross-language similarity in neural activation and which brain regions show the modulatory effects of language distance in orthographic transparency. To address these questions, the present study used representational similarity analysis (RSA) to precisely estimate the neural pattern similarity between native language and two non-native languages in Uyghur-Chinese-English trilinguals, whose third language (i.e., English) was more similar to the native language (i.e., Uyghur) in orthography than to their second language (i.e., Chinese). Behavioral results revealed that subjects responded faster to words in the non-native language with more similar orthography to their native language in the word naming task. More importantly, RSA revealed greater neural pattern similarity between Uyghur and English than between Uyghur and Chinese in select brain areas for phonological processing, especially in the left hemisphere. Further analysis confirmed that those brain regions represented phonological information. These results provide direct neuroimaging evidence for the modulatory effect of language distance in orthographic transparency on cross-language pattern similarity between native and non-native languages during word reading.

Detection of SFTS virus RNA and antibodies in severe fever with thrombocytopenia syndrome surveillance cases in endemic areas of China.

BACKGROUND: Severe fever with thrombocytopenia syndrome (SFTS) is a newly identified severe infectious disease caused by SFTS phlebovirus (SFTSV). SFTS monitoring has been carried out since 2010 in mainland China. We analysed the detection results of SFTSV RNA and antibody in SFTS surveillance cases to provide basic data for SFTS diagnosis. METHODS: This study was conducted in Shandong Province. Sera of SFTS surveillance cases were collected to detect SFTSV RNA and antibody by real-time RT-PCR and enzyme-linked immunosorbent assay, respectively. Detection rates were calculated. SPSS 18.0 (Chicago, IL, USA) was used for statistical analysis to compare the detection rates of SFTSV RNA and antibodies among different sera groups. RESULTS: A total of 374 SFTS surveillance cases were enrolled. Overall, 93.3% (349/374) of the sera samples were collected within 2 weeks after onset, and 6.7% (25/374) were collected between 15 days and 45 days. Of these, 183 (48.9%) were positive for SFTSV RNA. The SFTSV RNA-positive rate peaked (52.2%) in samples collected ≤7 days after onset and then showed a decreasing trend. The detection rate of SFTSV-specific IgM antibody was 30.5% (46/151) and was highest in samples collected between 8 and 14 days (43.3%, 26/60). The positive rate of SFTSV-specific IgG antibody (17.9%, 27/151) showed an increasing trend with the specimen collection time. In total, 74.8% (113/151) of sera samples had the same SFTSV RNA and IgM antibody detection results. However, 23.2% (29/125) of SFTSV RNA-negative cases were IgM antibody-positive, and 8.6% (9/105) of IgM antibody-negative cases were SFTSV RNA-positive. CONCLUSIONS: SFTSV RNA detection was preferred for SFTSV infection during disease surveillance. For highly suspected SFTS cases, IgM antibody is suggested to make a comprehensive judgement.

Reverse genetics system for severe fever with thrombocytopenia syndrome virus.

UNLABELLED: Severe fever with thrombocytopenia syndrome virus (SFTSV) is an emerging tick-borne pathogen that was first reported in China in 2009. Phylogenetic analysis of the viral genome showed that SFTS virus represents a new lineage within the Phlebovirus genus, distinct from the existing sandfly fever and Uukuniemi virus groups, in the family Bunyaviridae. SFTS disease is characterized by gastrointestinal symptoms, chills, joint pain, myalgia, thrombocytopenia, leukocytopenia, and some hemorrhagic manifestations with a case fatality rate of about 2 to 15%. Here we report the development of reverse genetics systems to study STFSV replication and pathogenesis. We developed and optimized functional T7 polymerase-based M- and S-segment minigenome assays, which revealed errors in the published terminal sequences of the S segment of the Hubei 29 strain of SFTSV. We then generated recombinant viruses from cloned cDNAs prepared to the antigenomic RNAs both of the minimally passaged virus (HB29) and of a cell culture-adapted strain designated HB29pp. The growth properties, pattern of viral protein synthesis, and subcellular localization of viral N and NSs proteins of wild-type HB29pp (wtHB29pp) and recombinant HB29pp viruses were indistinguishable. We also show that the viruses fail to shut off host cell polypeptide production. The robust reverse genetics system described will be a valuable tool for the design of therapeutics and the development of killed and attenuated vaccines against this important emerging pathogen. IMPORTANCE: SFTSV and related tick-borne phleboviruses such as Heartland virus are emerging viruses shown to cause severe disease in humans in the Far East and the United States, respectively. Study of these novel pathogens would be facilitated by technology to manipulate these viruses in a laboratory setting using reverse genetics. Here, we report the generation of infectious SFTSV from cDNA clones and demonstrate that the behavior of recombinant viruses is similar to that of the wild type. This advance will allow for further dissection of the roles of each of the viral proteins in the context of virus infection, as well as help in the development of antiviral drugs and protective vaccines.

Viral suppression function of intracellular antibody against C-terminal domain of rabies virus phosphoprotein.

Rabies virus (RV) causes a fatal disease in both human and animals. The disease can be prevented by post-exposure prophylaxis in individuals exposed to RV. However, the neutralization effect is limited after the virus enters into the host cells. So, it is important to identify new targets for rabies therapy. In this study, a human antibody RV1A2 specific to RV phosphoprotein (RV-P) was generated from a human naïve immune antibody library. The antibody recognized all forms of the phosphoproteins including the full length (P1) and short length of the P proteins (P2, P3, P4, and P5). The epitope mapping and the molecular docking of antigen-antibody complex showed that the antibody targets at a conserved epitope of 'VLGWV' ranging from amino acid (aa) 262 to 266 at C-terminal domain of the P protein, which locates at a hydrophobic pocket region in the C-terminal of the RV-P. The aa W265 within the epitope is on the flat surface of the domain, suggesting that it may be a critical amino acid for the functions of the P protein. Our results further showed that intracellular antibody RV1A2 which targets at the C-terminal domain of the P protein could effectively inhibit RV propagation 2-4 days post infection. These results suggest that the conserved C-terminal domain may be used as a new target for drug discovery, which highlights an intracellular inhibition of RV propagation and provides a potential novel way to treat RV infection.

Oral immunization with recombinant enterovirus 71 VP1 formulated with chitosan protects mice against lethal challenge.

BACKGROUND: Enterovirus 71 (EV71) is the etiologic agent of hand-foot-and-mouth disease (HFMD) in the Asia-Pacific region, Many strategies have been applied to develop EV71 vaccines but no vaccines are currently available. Mucosal immunization of the VP1, a major immunogenic capsid protein of EV71, may be an alternative way to prevent EV71 infection. RESULTS: In this study, mucosal immunogenicity and protect function of recombinant VP1 protein (rVP1) in formulation with chitosan were tested and assessed in female ICR mouse model. The results showed that the oral immunization with rVP1 induced VP1-specific IgA antibodies in intestine, feces, vagina, and the respiratory tract and serum-specific IgG and neutralization antibodies in vaccinated mice. Splenocytes from rVP1-immunized mice induced high levels of Th1 (cytokine IFN-γ), Th2 (cytokine IL-4) and Th3 (cytokine TGF-ß) type immune responses after stimulation. Moreover, rVP1-immunized mother mice conferred protection (survival rate up to 30%) on neonatal mice against a lethal challenge of 103 plaque-forming units (PFU) EV71. CONCLUSIONS: These data indicated that oral immunization with rVP1 in formulation with chitosan was effective in inducing broad-spectrum immune responses and might be a promising subunit vaccine candidate for preventing EV71 infection.

How the intrinsic functional connectivity patterns of the semantic network support semantic processing.

Semantic processing, a core of language comprehension, involves the activation of brain regions dispersed extensively across the frontal, temporal, and parietal cortices that compose the semantic network. To comprehend the functional structure of this semantic network and how it prepares for semantic processing, we investigated its intrinsic functional connectivity (FC) and the relation between this pattern and semantic processing ability in a large sample from the Human Connectome Project (HCP) dataset. We first defined a well-studied brain network for semantic processing, and then we characterized the within-network connectivity (WNC) and the between-network connectivity (BNC) within this network using a voxel-based global brain connectivity (GBC) method based on resting-state functional magnetic resonance imaging (fMRI). The results showed that 97.73% of the voxels in the semantic network displayed considerably greater WNC than BNC, demonstrating that the semantic network is a fairly encapsulated network. Moreover, multiple connector hubs in the semantic network were identified after applying the criterion of WNC > 1 SD above the mean WNC of the semantic network. More importantly, three of these connector hubs (i.e., the left anterior temporal lobe, angular gyrus, and orbital part of the inferior frontal gyrus) were reliably associated with semantic processing ability. Our findings suggest that the three identified regions use WNC as the central mechanism for supporting semantic processing and that task-independent spontaneous connectivity in the semantic network is essential for semantic processing.

Effects of short-term second language learning on the development of individual semantic networks in written and spoken language.

Previous studies have primarily focused on the relationship between native language (L1) and second language (L2) in the brain, specifically in one language modality, such as written or spoken language. However, there is limited research on how L2 proficiency impacts both modalities. This study aimed to investigate the functional networks involved in reading and speech comprehension for both L1 and L2, and observe changes in these networks as L2 proficiency improves. The dataset used in this study was obtained from a previous research conducted by Gurunandan et al., which involved Spanish-English bilingual participants undergoing a three-month English training program. Participants underwent fMRI scanning and performed a semantic animacy judgment task in both spoken and written language before and after training. Through analysis, distinct neural networks associated with spoken and written language were found between individuals' L1 and L2, both before and after training. Moreover, as L2 proficiency improved, the spoken and written networks for L2 remained distinct from those of the L1. These findings suggest that short-term L2 learning experiences can modify neural networks, but may not be enough to achieve native-like proficiency, supporting the accommodation hypothesis. These results have important implications for language learning and education, indicating that additional short-term training and exposure alone may not bridge the gap between L1 and L2 processing networks.

The Left Amygdala and Right Frontoparietal Cortex Support Emotional Adaptation Aftereffects.

Adaptation aftereffects-in which prolonged prior experience (adaptation) can bias the subsequent judgment of ambiguous stimuli-are a ubiquitous phenomenon. Numerous studies have found behaviorally stable adaptation aftereffects in a variety of areas. However, it is unclear which brain regions are responsible for this function, particularly in the case of high-level emotional adaptation aftereffects. To address this question, the present study used fMRI technology to investigate the neural mechanism of emotional adaptation aftereffects. Consistent with previous studies, we observed typical emotional adaptation effects in behavior. Specifically, for the same morphed facial images, participants perceived increased sadness after adapting to a happy facial image and increased happiness after adapting to a sad facial image. More crucially, by contrasting neural responses to ambiguous morphed facial images (i.e., facial images of intermediate morph levels) following adaptation to happy and sad expressions, we demonstrated a neural mechanism of emotional aftereffects supported by the left amygdala/insula, right angular gyrus, and right inferior frontal gyrus. These results suggest that the aftereffects of emotional adaptation are supported not only by brain regions subserving emotional processing but also by those subserving cognitive control.

Neural similarities and differences between native and second languages in the bilateral fusiform cortex in Chinese-English bilinguals.

In the field of bilingualism, researchers have proposed an assimilation hypothesis that posits that bilinguals apply the neural network of their native language to process their second language. In Chinese-English bilinguals, the bilateral fusiform gyrus has been identified as the key brain region showing the assimilation process. Specifically, in contrast to left-lateralized activation in the fusiform gyrus in native English speakers, Chinese-English bilinguals recruit the bilateral fusiform cortex to process English words as they do in the processing of Chinese characters. Nevertheless, it is unclear which type of information processing is assimilated in the fusiform gyrus. Using representational similarity analysis (RSA) and psychophysiological interaction (PPI) analysis, this study examined the differences in information representation and functional connectivity between both languages in the fusiform subregions in Chinese-English bilinguals. Univariate analysis revealed that both Chinese and English naming elicited strong activations in the bilateral fusiform gyrus, which confirmed the assimilation process at the activation intensity level. RSA indicated that the neural pattern of English phonological information was assimilated by Chinese in the anterior and middle right fusiform gyrus, while those of orthographic and visual form information were not. Further PPI analysis demonstrated that the neural representation of English phonological information in the right anterior fusiform subregion was related to its interaction with the frontotemporal areas for high-level linguistic processing, while the neural representation of English orthographic information in the right middle fusiform subregion was linked to its interaction with the left inferior occipital cortex for visual processing. These results suggest that, despite the recruitment of similar neural resources in one's native and second languages, the assimilation of information representation is limited in the bilateral fusiform cortex. Our results shed light on the neural mechanisms of second language processing.

Category-specific and category-general neural codes of recognition memory in the ventral visual pathway.

Researchers have identified category-specific brain regions, such as the fusiform face area (FFA) and parahippocampal place area (PPA) in the ventral visual pathway, which respond preferentially to one particular category of visual objects. In addition to their category-specific role in visual object identification and categorization, regions in the ventral visual pathway play critical roles in recognition memory. Nevertheless, it is not clear whether the contributions of those brain regions to recognition memory are category-specific or category-general. To address this question, the present study adopted a subsequent memory paradigm and multivariate pattern analysis (MVPA) to explore category-specific and category-general neural codes of recognition memory in the visual pathway. The results revealed that the right FFA and the bilateral PPA showed category-specific neural patterns supporting recognition memory of faces and scenes, respectively. In contrast, the lateral occipital cortex seemed to carry category-general neural codes of recognition memory. These results provide neuroimaging evidence for category-specific and category-general neural mechanisms of recognition memory in the ventral visual pathway.

Knockout of CLTC gene reduces but not completely block SFTSV infection.

Clathrin is a key protein for viruses to enter host cells. Previous studies often use clathrin inhibitors or gene knockdown technology to partially inhibit the function of clathrin, but whether SFTSV can infect host cells without clathrin expression remains unclear. In this research, a clathrin heavy chains (CLTC) knockout A549 cell line was established by CRISPR/Cas9 technology, and the knockout of CLTC was verified by PCR, Western blot, immunofluorescence and T7E1 analysis. The off-target effect was evaluated by PCR combined with Sanger sequencing. Furthermore, this research verified that SFTSV infection was significantly inhibited, but not completely blocked, due to the deletion of CLTC protein. Our research also found that lipid raft inhibitor Filipin, other than macropinocytosis inhibitor EIPA, could significantly reduce SFTSV infection, and the inhibition was more obviously observed when Filipin was used in CLTC knockout cells. These result indicated that clathrin-dependent and lipid raft mediated endocytosis are the major two mode used by SFTSV entry. In conclusion, this study constructed a CLTC knockout cell line, which, for the first time, established a cell model for the study of the function of CLTC protein, and provided direct evidence that SFTSV pendent could still infect cells without clathrin. Additionally, we confirmed that lipid raft mediated endocytosis, as a clathrin-independent pathway, could be another key mode for SFTSV entry.

Optimization of differentiation and transcriptomic profile of THP-1 cells into macrophage by PMA.

THP-1 monocyte, which can be differentiated into macrophages by PMA, is widely used in researches on pathogen infection and host innate immunity, but reports on the induction methods of PMA are different and lack a unified standard, and the transcriptome characteristics of macrophage compared with THP-1 cells remains unclear. In this research, we examined the differentiation effect of three factors including induction time, cell seeding density and PMA concentration by detecting the positive rate of CD14 expression. The concentration of 80ng/ml of PMA, the induction time of 24h, and the cell seeding density of 5×105 cells/ml, could respectively facilitates a relatively higher CD14 positive rate in THP-1 cells. Under this optimized conditions, the CD14 positive rate of THP-1 cells can reach 66.52%. Transcriptome sequencing showed that after the above induction, the mRNA expression of 3113 genes which were closely related to cell communication, signal transduction, cell response to stimulus, signaling receptor binding and cytokine activity were up-regulated, and the top 10 genes were RGS1, SPP1, GDF15, IL-1B, HAVCR2, SGK1, EGR2, TRAC, IL-8 and EBI3. While the mRNA expression of 2772 genes which were associated with cell cycle process, DNA binding and replication and cell division, were down-regulated, and the top genes were SERPINB10, TRGC2, SERPINB2, TRGC1, MS4A3, MS4A4E, TRGJP1, MS4A6A, TRGJP2, MS4A4A. This research optimized the induction method on THP-1 cell differentiation from three aspects and delineated the transcriptomic profile of PMA-induced THP-1 cells, laying a foundation for the construction method of cell model and for the functional study of macrophage.

Co-circulation and co-infection of hantaviruses and Wenzhou mammarenavirus in small mammals and humans in Jiangxi, China.

Both Orthohantaviruses (HV) and Whenzhou Mammarenaviruses (WENV) are rodents borne viruses, allowing them to spread simultaneously in the same area and infect humans. To explore the potential threat of HV and WENV to public health safety, an environmental and laboratory investigation was conducted in 2020-2021, in Jiangxi province, China. A total of 461 small mammals of 7 species and paired sera from 43 suspected HFRS cases were collected from Jiangxi Province, China. Viral genomic RNA and specific antibodies against HV and WENV were detected to evaluate the epidemic situation of the two viruses. Hantaan virus (HTNV), seoul virus (SEOV) and WENV RNA were detected in the lungs of the captured mammals, which resulted 4.1% and 7.4% of HV and WENV RNA positive respectively. Co-infections of WENV and SEOV were detected from Rattus norvegicus, Mus musculus and Rattus flavipectus with an overall co-infection rate of 0.65%. The detection rates of antibodies in the blood against HV and WENV were 11.9% (55/461), and 13.2% (61/461) respectively. The prevalence of viral infection and viral genetic characters varied among the selected areas. In the paired sera of 43 suspected HFRS cases, 38 were with HV infection, 11 were with WENV IgG, and 7 with a 4-fold or more of WENV IgG titer elevation. These results revealed the fact of the co-circulating and coinfection of HV and WENV in the same area at the same time, which might impact on public health safety.

Research on the Effect of Water Injection Pressure on Knocking Combustion of a Direct Injection Gasoline Engine.

In this study, the research method of numerical simulation is used to explore the inhibition of different water injection pressures on knock combustion of turbocharged direct injection gasoline (GDI) engines by coupling computational fluid dynamics with a chemical-kinetics model. First, the ignition advance angle and compression ratio are increased to induce the GDI engine to knock, and then the influence of the water injection pressure on the in-cylinder, evaporation of water, and the knock of the gasoline engine are analyzed. The simulation results show that, compared with no water injection, the direct injection of water in the cylinder can significantly reduce the knock intensity. When the water injection pressure is greater than 40 bar, the knock intensity is less than 2 and the knocking is completely suppressed. In this work, the effects of different water injection pressures on knocking are explored by analyzing the effects of water injection pressure on water atomization, in-cylinder combustion, and the knocking mechanism. On the one hand, the evaporation rate of water increases with increasing water injection pressure and the quality of the liquid film generally improves. On the other hand, direct water injection can significantly reduce the distribution of CH2O in the end mixture, thereby reducing the generation of H2O2 and further suppressing the spontaneous combustion of the end mixture. At the moment of knock, when the water injection pressure is greater than 40 bar, the detonation mechanism of the no. 7 monitoring point does not produce a sudden change in HCO radicals. The water spray can effectively reduce the NO x emission, and the NO x emission under the water spray pressure of 120 bar is the lowest. However, after spraying water, it will increase CO emissions.

Individuals' preference on reading pathways influences the involvement of neural pathways in phonological learning.

Introduction: Existing behavioral and neuroimaging studies revealed inter-individual variability in the selection of the two phonological routes in word reading. However, it is not clear how individuals' preferred reading pathways/strategies modulate the involvement of a certain brain region for phonological learning in a new language, and consequently affect their behavioral performance on phonological access. Methods: To address this question, the present study recruited a group of native Chinese speakers to learn two sets of artificial language characters, respectively, in addressed-phonology training (i.e., whole-word mapping) and assembled-phonology training conditions (i.e., grapheme-to-phoneme mapping). Results: Behavioral results showed that the more lexical pathways participants preferred, the better they performed on newly-acquired addressed characters relative to assembled characters. More importantly, neuroimaging results showed that participants who preferred lexical pathway in phonological access show less involvement of brain regions for addressed phonology (e.g., the bilateral orbitofrontal cortex and right pars triangularis) in the processing of newly-acquired addressed characters. Conclusion: These results indicated that phonological access via the preferred pathway required less neural resources to achieve better behavioral performance. These above results provide direct neuroimaging evidence for the influence of reading pathway preference on phonological learning.

CRISPR/Cas12a Technology Combined With RPA for Rapid and Portable SFTSV Detection.

Severe fever with thrombocytopenia syndrome virus (SFTSV) is a new tick-borne pathogen that can cause severe hemorrhagic fever. Fever with thrombocytopenia syndrome caused by SFTSV is a new infectious disease that has posed a great threat to public health. Therefore, a fast, sensitive, low-cost, and field-deployable detection method for diagnosing SFTSV is essential for virus surveillance and control. In this study, we developed a rapid, highly sensitive, instrument-flexible SFTSV detection method that utilizes recombinase polymerase amplification and the CRISPR/Cas12a system. We found that three copies of the L gene from the SFTSV genome per reaction were enough to ensure stable detection within 40 min. The assay clearly showed no cross-reactivity with other RNA viruses. Additionally, our method demonstrated 100% agreement with Q-PCR detection results for SFTSV in 46 clinical samples. We simplified the requirements for on-site detection instruments by combining the CRISPR/Cas12a tool and immunochromatographic strips to create a system that can reliably detect one copy/µl sample of the L gene, which showed extremely high sensitivity and specificity for detecting the virus. Taken together, these findings indicate that the new SFTSV detection method is a powerful and effective tool for on-site detection, which can contribute to diagnosing SFTSV quickly and sensitively.

Development and evaluation of recombinant E2 protein based IgM capture enzyme-linked immunosorbent assay (ELISA) and double antigen sandwich ELISA for detection of antibodies to Chikungunya virus.

BACKGROUND: Chikungunya virus (CHIKV) reemerged and caused millions of human infections since 2004. The disease could be established, when the virus has been introduced to areas where the appropriate vectors are endemic. The differential diagnosis of CHIKV infection varies based on place of residence, travel history, and exposures. Serological tests are commonly used to diagnose CHIKV infection, but their availability and assessments of the performance of the diagnostics have been limited. OBJECTIVES: To develop and evaluate antibodies detection methods for chikungunya diagnosis and serological investigation. METHODS: Recombinant E2 protein based IgM capture enzyme-linked immunosorbent assay (Mac-ELISA) and double antigen sandwich ELISA (Das-ELISA) for detection of antibodies to Chikungunya virus were developed and evaluated. The repeatability was evaluated by testing of three reference sera at single dilutions in triplicated for 5 times. The sensitivity, specificity, accuracy, and agreement of the MAC-ELISA and Das-ELISA were obtained by comparing the detection results of 225 serum samples (45 positive; 180 negative) with a real-time RT-PCR assay and an IFA commercial tests manufactured by Euroimmun. RESULTS: The established ELISA assays were standardized by determining the optimal concentrations of the key reagents. The coefficient values of repeat testing were within 10% and 20% for intraassay and interassay precision, respectively. A sensitivity of 60.0% and 52.5%, a specificity of 96.2% and 96.8%, and an accuracy of 89.8% and 88.9% were obtained for the Mac-ELISA and Das-ELISA, respectively, when compared to a CHIKV qRT-PCR method. And a sensitivity of 100%, a specificity of 97.5% and 99.5%, and an accuracy of 97.8% and 99.6% were yielded respectively when using the IIFT as a reference method, which showed a highly consistence to the commercial IIFT assay with a Kappa value greater than 0.90. CONCLUSIONS: The Mac-ELISA and Das-ELISA based on recombinant E2 protein of CHIKV were developed and standardized, which could detect IgM or total antibodies against CHIKV in 2-3 hours with acceptable sensitivities and specificities. These assays can be used for laboratory diagnosis and serological investigation of CHIKV infections to evaluate the risk of CHIKV transmission.

A Simulation Study of Water Injection Position and Pressure on the Knock, Combustion, and Emissions of a Direct Injection Gasoline Engine.

A turbocharged downsizing spark ignition (SI) engine combined with direct injection technology has the potential to improve the power and fuel economy and reduce emissions. However, gasoline engines are prone to knocking under low-speed and high-load conditions, which limits the application and development of downsizing SI engines. In this study, numerical simulation methods are used to explore the feasibility of water injection in the intake port to reduce the knock tendency of gasoline direct injection (GDI) engines and to explore the effects of different water injection pressures on combustion and emissions. First, the GDI engine is induced to knock by increasing the compression ratio and advancing the spark timing. Then, the influences of low position and no angle (LPNA) and high position and angled water injector arrangements on engine combustion are explored. When the water injector arrangement is LPNA, the turbulent kinetic energy near the spark plug is higher, the equivalence ratio is more evenly distributed, and the engine knock intensity is smaller. Finally, when the arrangement of the water injector is LPNA, the effects of water injection pressure on the knock, combustion, and emissions of the GDI engine are explored. The results show that when the water injection pressure is 5 bar, the knock intensity of the engine is the smallest, the cycle work is the highest, and the emissions of NO x and unburned hydrocarbon are the lowest.