A duplex real-time reverse transcriptase polymerase chain reaction assay for detecting western equine and eastern equine encephalitis viruses

In order to establish an accurate, ready-to-use assay for simultaneous detection of Eastern equine encephalitis virus (EEEV) and Western equine encephalitis virus (WEEV), we developed one duplex TaqMan real-time reverse transcriptase polymerase chain reaction (RT-PCR) assay, which can be used in human and vector surveillance. First, we selected the primers and FAM-labeled TaqMan-probe specific for WEEV from the consensus sequence of NSP3 and the primers and HEX-labeled TaqMan-probe specific for EEEV from the consensus sequence of E3, respectively. Then we constructed and optimized the duplex real-time RT-PCR assay by adjusting the concentrations of primers and probes. Using a series of dilutions of transcripts containing target genes as template, we showed that the sensitivity of the assay reached 1 copy/reaction for EEEV and WEEV, and the performance was linear within the range of at least 10(6 )transcript copies. Moreover, we evaluated the specificity of the duplex system using other encephalitis virus RNA as template, and found no cross-reactivity. Compared with virus isolation, the gold standard, the duplex real time RT-PCR assay we developed was 10-fold more sensitive for both WEEV and EEEV detection

Comprehensive Mapping of Common Immunodominant Epitopes in the West Nile Virus Nonstructural Protein 1 Recognized by Avian Antibody Responses

West Nile virus (WNV) is a mosquito-borne flavivirus that primarily infects birds but occasionally infects humans and horses. Certain species of birds, including crows, house sparrows, geese, blue jays and ravens, are considered highly susceptible hosts to WNV. The nonstructural protein 1 (NS1) of WNV can elicit protective immune responses, including NS1-reactive antibodies, during infection of animals. The antigenicity of NS1 suggests that NS1-reactive antibodies could provide a basis for serological diagnostic reagents. To further define serological reagents for diagnostic use, the antigenic sites in NS1 that are targeted by host immune responses need to be identified and the potential diagnostic value of individual antigenic sites also needs to be defined. The present study describes comprehensive mapping of common immunodominant linear B-cell epitopes in the WNV NS1 using avian WNV NS1 antisera. We screened antisera from chickens, ducks and geese immunized with purified NS1 for reactivity against 35 partially overlapping peptides covering the entire WNV NS1. This study identified twelve, nine and six peptide epitopes recognized by chicken, duck and goose antibody responses, respectively. Three epitopes (NS1-3, 14 and 24) were recognized by antibodies elicited by immunization in all three avian species tested. We also found that NS1-3 and 24 were WNV-specific epitopes, whereas the NS1-14 epitope was conserved among the Japanese encephalitis virus (JEV) serocomplex viruses based on the reactivity of avian WNV NS1 antisera against polypeptides derived from the NS1 sequences of viruses of the JEV serocomplex. Further analysis showed that the three common polypeptide epitopes were not recognized by antibodies in Avian Influenza Virus (AIV), Newcastle Disease Virus (NDV), Duck Plague Virus (DPV) and Goose Parvovirus (GPV) antisera. The knowledge and reagents generated in this study have potential applications in differential diagnostic approaches and subunit vaccines development for WNV and other viruses of the JEV serocomplex

Wheat TaRab7 GTPase Is Part of the Signaling Pathway in Responses to Stripe Rust and Abiotic Stimuli

Small GTP-binding proteins function as regulators of specific intercellular fundamental biological processes. In this study, a small GTP-binding protein Rab7 gene, designated as TaRab7, was identified and characterized from a cDNA library of wheat leaves infected with Puccinia striiformis f. sp. tritici (Pst) the wheat stripe rust pathogen. The gene was predicted to encode a protein of 206 amino acids, with a molecular mass of 23.13 KDa and an isoeletric point (pI) of 5.13. Further analysis revealed the presence of a conserved signature that is characteristic of Rab7, and phylogenetic analysis demonstrated that TaRab7 has the highest similarity to a small GTP binding protein gene (BdRab7-like) from Brachypodium distachyon. Quantitative real-time PCR assays revealed that the expression of TaRab7 was higher in the early stage of the incompatible interactions between wheat and Pst than in the compatible interaction, and the transcription level of TaRab7 was also highly induced by environmental stress stimuli. Furthermore, knocking down TaRab7 expression by virus induced gene silencing enhanced the susceptibility of wheat cv. Suwon 11 to an avirulent race CYR23. These results imply that TaRab7 plays an important role in the early stage of wheat-stripe rust fungus interaction and in stress tolerance

Filozoficzne refleksje nad posthumanizmem w utworze Stanisława Lema Golem XIV

Stanisław Lem był wielkim pisarzem, futurystą i krytykiem literackim, ale chińskie przekłady większości jego dzieł zostały wydane w Chinach dopiero z okazji setnej rocznicy urodzin autora w 2021 roku. Z tej przyczyny prowadzone w Chinach badania naukowe na temat Lema i jego twórczości są nieliczne. Wielu czytelników nie do końca rozumie też idee zawarte w książkach Stanisława Lema. Istnieje zatem potrzeba znalezienia odpowiednich metod analizy i badania twórczości pisarza. W niniejszym artykule wykazano, że perspektywa posthumanistyczna może dostarczyć wiarygodnego wzoru dla studiów nad Lemem. W latach 90. XX wieku popularność zdobyły interdyscyplinarne badania nad posthumanizmem, a ich koncepcja personifikacji technologii i „cyborga” otworzyła szeroką przestrzeń pisarzom reprezentującym literaturę popularnonaukową. Jednak już w latach 70. Lem złożył deklarację posthumanizmu, w której wykreował typową postać cyborga – Golema XIV – i przez narrację tej sztucznej inteligencji wyraził własne poglądy na temat relacji między człowiekiem a technologią, człowiekiem a nie-człowiekiem. W niniejszym artykule przeanalizowano dzieło Lema Golem XIV. Przedstawione w tym utworze filozoficzne idee autora nie tylko pokrywają się pod wieloma względami z dzisiejszymi teoriami posthumanistycznymi, lecz także wyrażają pewne wnikliwe refleksje, które jako bardziej futurystyczne wykraczają daleko poza ramy myśli posthumanistycznej. Lem skupia się przede wszystkim na interpretacji ewolucji i mądrości. Cel ludzkiego istnienia wyjaśnia w kategoriach ewolucji, co mieści się w ramach posthumanizmu. Jednak prezentowany utwór zawiera nie tylko posthumanistyczną krytykę antropocentryzmu – także dopełnia dekonstrukcję człowieka. Wielu badaczy we wcześniejszych opracowaniach łączyło twórczość Lema z posthumanizmem, ale opracowaniom tym brakowało teoretycznego wsparcia. Niewiele też mówiono o historii rozwoju dyscyplinarnego tego nurtu, co uczyniono w niniejszym artykule. Zawarto w nim ponadto omówienie kompozycji układu treści dzieła Lema Golem XIV, przedstawiono pogłębioną analizę obecnych w nim idei posthumanistycznych oraz wydestylowano unikalne poglądy filozoficzne Lema w nadziei na dostarczenie nowych pomysłów na interpretację jego utworu.In the 1990s, posthumanism gained popularity in interdisciplinary research, and its concept of the personification of technology and the “cyborg” offered a wide scope for science fiction. But as early as the 1970s, Polish science fiction novelist Stanisław Lem had already made a manifesto of posthumanism in his work, in which he created Golem XIV, a typical cyborg, and expressed his own views on the relationship between human and technology, human and non-human, etc. through the narration of this artificial intelligence. In his book, he created Golem XIV, the archetypal cyborg. The philosophical ideas of Lem in Golem XIV not only coincide with the present theories of posthumanism in many ways, but also have some insightful thoughts that go far beyond the framework of posthumanist thought and are more forward-looking. This paper includes the history of the development of the discipline of posthumanism, discussion on the content of Lem’s Golem XIV, an in-depth analysis of posthumanist thinking, and Lem’s own unique philosophical views in the hope of providing new ideas for interpreting Lem’s work

Experimental Investigation of the Dynamic Response of a Sliding Bearing System under Different Oil Pressure Levels

A sliding bearing system performs a critical function in a rotating machinery system and provides support for the rotating components. In this paper, a single-rotor experimental rig and a dual-rotor sliding bearing experimental rig were independently established, and the dynamic response of the test systems are investigated. To study the effect of oil pressure levels on the dynamic response of the test systems under the single-rotor system and the dual-rotor system, the vibration response of single-rotor and dual-rotor systems ware analyzed by different oil inlet pressures to study the effect of oil pressure levels on the dynamic response of the test systems under the single-rotor system and the dual-rotor system. Among them, the single rotor system under a large oil pressure system is mostly periodic and more stable. While under different inlet oil pressure conditions applied in the experiments, the dynamic motion of the dual-rotor bearing system is relatively stable in both the subcritical and supercritical speed regions. However, the oil whirl and oil whip of the dual-rotor system were observed at the critical speed, which causes the system into unstable multiperiodic motions. In general, this paper can give some insight into the dynamic response of the sliding bearing system influenced by different oil pressure levels

MicroNano2008-70076 INVESTIGATION ON MULTIPLE PIEZOELECTRIC EFFECT OF PIEZOELECTRIC CRYSTAL UNDER APPLIED ELECTRIC FIELD

ABSTRACT Based on the theory of classical piezoelectricity, the relationship between the generation of multiple piezoelectric effect under applied electric field and boundary conditions is analyzed. The generating process of multiple piezoelectric effect is elaborated and the formulas of secondary direct and tertiary converse piezoelectric effect are deduced. The displacement or voltage generated by primary converse, secondary direct and tertiary converse piezoelectric effect is calculated by using coupled-field analysis function in the Finite Element Analysis software-ANSYS. Experimental research of multiple piezoelectric effect under applied electric field is performed through PZT-5 stack. The experimental result verifies the existence of multiple piezoelectric effect. Experimental data shows that voltage and displacement generated by secondary direct and tertiary converse piezoelectric effect respectively are linear with the applied voltage. Both the simulation and experimental results prove that the theoretical analysis is accurate. Keywords: multiple piezoelectric effect, boundary condition, PZT-5 stack, MEMS INTRODUCTION Since piezoelectricity was discovered, many scientists have investigated extensively piezoelectric effect. At present, the study of piezoelectric effect has already expanded from 2-dimensional effect based on stress analysis, namely the longitudinal effect, transverse effect and sheer effect to the 3-dimensional effect based on the deformation analysis, such as the extending effect, compressing effect, bending effect and torsional effect By using the knowledge of interdisciplines such as Piezoelectricity, Physics of Crystals, Physics of Dielectrics and Theory of Elasticity, the relationship between the generation of multipl

Preparation of Imazethapyr Surface Molecularly Imprinted Polymers for Its Selective Recognition of Imazethapyr in Soil Samples

A novel strategy based on imazethapyr (IM) molecular-imprinting polymers (MIPs) grafted onto the surface of chloromethylation polystyrene resin via surface-initiated atom transfer radical polymerization (SI-ATRP) for specific recognition and sensitive determination of trace imazethapyr in soil samples was developed. The SI-ATRP was performed by using methanol-water (4 : 1, v/v) as the solvent, acrylamide as the functional monomer, trimethylolpropane trimethacrylate (TRIM) as the cross-linker, imazethapyr as the template, and CuBr/2,2′-bipyridine as the catalyst. The resulting MIPs were characterized by elemental analysis, Fourier-transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM), and transmission electron microscopy (TEM). Then, the binding selectivity, adsorption capacity, and reusability of the MIPs were evaluated. The results indicated that the prepared MIPs exhibited specific recognition and high selectivity for imazethapyr. The MIPs were further used as solid-phase extraction (SPE) materials coupled with high-performance liquid chromatography (HPLC) for selective extraction and detection of trace imazethapyr from soil samples. The results showed that good linearity was observed in the range of 0.10–5.00 μg/mL, with a correlation coefficient of 0.9995. The limit of detection (LOD) of this method was 15 ng/g, and the extraction recoveries of imazethapyr from real samples were in the range of 91.1–97.5%, which proved applicable for analysis of trace imazethapyr in soils. This work proposed a sensitive, rapid, and convenient approach for determination of trace imazethapyr in soil samples

Nuclear protein 1 promotes unfolded protein response during endoplasmic reticulum stress, and alleviates apoptosis induced by cisplatin in non-small cell lung cancer cells

Purpose: To investigate the role of nuclear protein 1 (NUPR1) in the drug resistance of non-small cell lung cancer (NSCLC) and its regulatory mechanisms. Methods: Quantitative polymerase chain reaction (qPCR) and immunoblot assays were conducted to determine NUPR1 expression in A549 cells. Cisplatin sensitivity and cisplatin-induced apoptosis were investigated in NUPR1 knockdown or overexpressed cells via 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide (MTT) assay and flow cytometry. The potential association between unfolded protein response (UPR) and NUPR1 levels in response to cisplatin were explored. The effect of endoplasmic reticulum (ER) stress on apoptosis was examined using flow cytometry. Results: Cisplatin treatment promoted the expression of NUPR1 in NSCLC cells. NUPR1 regulated cisplatin resistance in NSCLC and also regulated UPR in ER stress induced by cisplatin. The results show NUPR1 regulated apoptosis induced by ER stress following tunicamycin treatment. Conclusion: NSCLC cells may promote the UPR in ER stress by promoting the expression of NUPR1, thereby reducing the ER stress induced by cisplatin

U4 at the 3' UTR of PB1 segment of H5N1 influenza virus promotes RNA polymerase activity and contributes to viral pathogenicity.

The viral RNA-dependent RNA polymerase has been found to contribute to efficient replication in mammalian systems and to the high pathogenicity of H5N1 influenza A virus in humans and other mammals. The terminal untranslated regions of the viral segments perform functions such as polyadenylation and contain signals for genomic packaging and initiation of RNA synthesis. These sequences are highly conserved, apart from a U/C polymorphism at position 4 of the 3' end, most often seen in the polymerase gene segments. However, no study has yet tested whether the untranslated regions of H5N1 make any contribution to its high pathogenicity. Herein, the association of the fourth nucleotide at the 3' end of the untranslated region in segment 2 (PB1), of A/Vietnam/1194/2004 (H5N1), with pathogenicity was examined in mice. To this end, an RNA polymerase reporter system was constructed, and viruses with mutations at this site were rescued. Results showed the U4 in PB1 was found to contribute to greater amounts of RNA-dependent RNA polymerase activity and differentially regulate genomic transcription and replication. Although a recombinant H5N1 virus with the rarer C4 sequence in all eight segments was viable and replicated to high titers in vitro, replacing a single U4 at the 3' termini of the PB1 gene segment enhanced viral reproduction and more pathogenesis. In this way, these data showed the importance of untranslated regions of H5N1 influenza virus to pathogenicity

Conductive Porous MXene for Bionic, Wearable, and Precise Gesture Motion Sensors

Reliable, wide range, and highly sensitive joint movement monitoring is essential for training activities, human behavior analysis, and human-machine interfaces. Yet, most current motion sensors work on the nano/microcracks induced by the tensile deformation on the convex surface of joints during joint movements, which cannot satisfy requirements of ultrawide detectable angle range, high angle sensitivity, conformability, and consistence under cyclic movements. In nature, scorpions sense small vibrations by allowing for compression strain conversion from external mechanical vibrations through crack-shaped slit sensilla. Here, we demonstrated that ultraconformal sensors based on controlled slit structures, inspired by the geometry of a scorpion’s slit sensilla, exhibit high sensitivity (0.45%deg-1), ultralow angle detection threshold (~15°), fast response/relaxation times (115/72 ms), wide range (15° ~120°), and durability (over 1000 cycles). Also, a user-friendly, hybrid sign language system has been developed to realize Chinese and American sign language recognition and feedback through video and speech broadcasts, making these conformal motion sensors promising candidates for joint movement monitoring in wearable electronics and robotics technology