177 research outputs found

    Soil aggregate size distribution and total organic carbon in intra-aggregate fractions as affected by addition of biochar and organic amendments

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    A two-year field trial on maize (Zea mays L.) production was established to determine the influence of biochar, maize straw, and poultry manure on soil aggregate stability, aggregate size distribution, total organic carbon (TOC), and soil microbial biomass carbon (MBC). Seven treatments with four replications, namely CK, control; S, 12.5 Mg ha-1 straw; B1, 12.5 Mg ha-1 biochar; B2, 25 Mg ha-1 biochar; SB1, straw + 12.5 Mg ha-1 biochar; SB2, straw + 25 Mg ha-1 biochar; and M, 25 Mg ha-1 manure were tested at four soil depths (0–10, 10–20, 20–30, and 30–40 cm). Aggregates were grouped into large macro-aggregates (5–2 mm), small macro-aggregates (2–0.25 mm), micro-aggregates (0.25–0.053 mm) and silt + clay (<0.053 mm). Biochar, straw, and manure applications all had significant effects (p < 0.05) on aggregate stability, with B2 at 20 cm soil depth showing the greatest increase (62.1%). SB1 of small macro-aggregate fraction showed the highest aggregate proportion (50.59% ± 10.48) at the 20–30 cm soil depth. The highest TOC was observed in SB2  (40.9 g kg-1) of large macro-aggregate at 10–20 cm soil depth. Treatment effects on soil MBC was high, with B1 showing the greatest value (600.0 µg g-1) at the 20–30 cm soil depth. Our results showed that application of biochar, straw, and manure to soil increased aggregate stability, TOC as well as MBC

    Candida albicans Cdc37 interacts with the Crk1 kinase and is required for Crk1 production

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    AbstractCrk1, a Cdc2-related protein kinase from the human pathogenic fungus Candida albicans, plays an important role in hyphal development and virulence. To address its regulatory mechanisms, we searched for Crk1 interacting proteins by two-hybrid screening. A CDC37 ortholog (CaCDC37) was cloned from the screening with the Crk1 kinase domain as the bait. The CaCdc37 interacted preferentially with the kinase domain of Crk1 (Crk1N) as shown by two-hybrid and immunoprecipitation experiments. CaCDC37 could complement a cdc37 thermosensitive mutant (cdc37-34) of Saccharomyces cerevisiae. Importantly, Crk1 protein was hardly detectable in the cdc37-34 mutant at restrictive temperature. However, upon expression of CaCdc37 in the cdc37 mutant, Crk1 protein was detected even at restrictive temperature. Our data suggested that CaCdc37 was required for the production of Crk1 kinase. Like Cdc37 proteins of S. cerevisiae and higher eukaryotes, CaCdc37 might function as a molecular chaperone that stabilized Crk1 and other protein kinases in C. albicans. In support of this, CaSTI1 was identified from a two-hybrid screen with the full-length Crk1 as the bait. CaSti1 showed two-hybrid interactions with both Crk1 and the CaCdc37

    BRIEF LIFE, ETERNAL MEMORY: PUSHKIN STUDIES IN CHINA

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    Pushkin is among the writers whose works remain relevant in many countries for many decades. In China, the interest of readers and researchers in the poet’s biography and his artistic world has been steadily increasing since the moment Pushkin’s works first appeared there. This article considers three stages in the study of Pushkin’s biography and works in China. The research shows how the perception of Pushkin’s works depends on the historical and sociocultural situation. The article covers works by key Chinese researchers. The author analyzes specific features of methodological approaches to studying Pushkin’s writings in a wide range of works by Chinese scholars, and shows the main vectors in the development of methodology of literary studies in the 20th century, and especially at the present stage of the humanities development. The article also shows how the changes in Chinese culture over the last hundred years influenced the formation and functioning of the image of Pushkin, as well as perception of his works in China. The author of the article comes to the conclusion that these changes have been caused by historical and socio-cultural shifts. Special attention is paid to the contemporary stage in the study of Pushkin's works by Chinese researchers of Russian culture and literature

    Clinical Application of Exosome Components

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    Exosomes belong to a subpopulation of EVs that carry different functional molecular cargoes, including proteins, nucleic acids, metabolites, and lipids. Notably, evidence has demonstrated that exosomes participate in bidirectional cell–cell communication and act as critical molecular vehicles in regulating numerous physiological and pathological processes. Since the specific contents within exosomes carry the information from their cells of origin, this property permits exosomes to act as valuable biomarkers. This chapter summarizes the potential use of exosome components in diagnosing, prognosis, or monitoring and treating multiple cancers and other non-neoplastic diseases. We also discuss the deficiency of basic applications, including the limitations of research methods and different research institutions and the differences generated by specimen sources. Thus, a better understanding of the problem of exosome detection may pave the way to promising exosome-based clinical applications

    Quantitative Polymerase Chain Reaction Coupled With Sodium Dodecyl Sulfate and Propidium Monoazide for Detection of Viable Streptococcus agalactiae in Milk

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    Streptococcus agalactiae is an important pathogen causing bovine mastitis. The aim of this study was to develop a simple and specific method for direct detection of S. agalactiae from milk products. Propidium monoazide (PMA) and sodium dodecyl sulfate (SDS) were utilized to eliminate the interference of dead and injured cells in qPCR. Lysozyme (LYZ) was adopted to increase the extraction efficiency of target bacteria DNA in milk matrix. The specific primers were designed based on cfb gene of S. agalactiae for qPCR. The inclusivity and exclusivity of the assay were evaluated using 30 strains. The method was further determined by the detection of S. agalactiae in spiked milk. Results showed significant differences between the SDS–PMA–qPCR, PMA–qPCR and qPCR when a final concentration of 10 mg/ml (R2 = 0.9996, E = 95%) of LYZ was added in DNA extraction. Viable S. agalactiae was effectively detected when SDS and PMA concentrations were 20 μg/ml and 10 μM, respectively, and it was specific and more sensitive than qPCR and PMA–qPCR. Moreover, the SDS–PMA–qPCR assay coupled with LYZ was used to detect viable S. agalactiae in spiked milk, with a limit of detection of 3 × 103 cfu/ml. Therefore, the SDS–PMA–qPCR assay had excellent sensitivity and specificity for detection of viable S. agalactiae in milk

    The Pichia pastoris transmembrane protein GT1 is a glycerol transporter and relieves the repression of glycerol on AOX1 expression

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    Promoter of alcohol oxidase I (PAOX1) is the most efficient promoter involved in the regulation of recombinant protein expression in Pichia pastoris (P. pastoris). PAOX1 is tightly repressed by the presence of glycerol in the culture medium; thus, glycerol must be exhausted before methanol can be taken up by P. pastoris and the expression of the heterologous protein can be induced. In this study, a candidate glycerol transporter (GT1, GeneID: 8197545) was identified, and its role was confirmed by further studies (e.g. bioinformatics analysis, heterologous complementation in Schizosaccharomyces pombe (S. pombe)). When GT1 is co-expressed with enhanced green fluorescent protein (EGFP), it localizes to the membrane and S. pombe carrying gt1 but not the wild-type strain can grow on medium containing glycerol as the sole carbon source. The present study is the first to report that AOX1 in the X-33gt1 mutant can achieve constitutive expression in medium containing glycerol; thus, knocking down gt1 can eliminate the glycerol repression of PAOX1 in P. pastoris. These results suggest that the glycerol transporter may participate in the process of PAOX1 inhibition in glycerol medium

    Correlation Between Protein Primary Structure and Soluble Expression Level of HSA dAb in Escherichia coli

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    Izoelektrična točka, duljina molekule, molekularna masa i slijed aminokiselina bitno utječu na topljivost proteina. U ovom smo se radu fokusirali na sastav aminokiselina i ispitali one koje najviše utječu na razinu ekspresije topljivog protutijela albumina iz ljudskog seruma (HSA dAb). Grupiranjem i primjenom linearnog modela analizirana je topljivost 65 varijanti proteina. Bitan utjecaj na ekspresiju topljivog protutijela dAb imale su specifične kombinacije aminokiselina, i to (S, R, N, D, Q) u supernatantu, (G, R, C, N, S) u lizatu peleta i (R, S, G) u ukupnom topljivom protutijelu dAb. Od 20 aminokiselina, arginin je imao negativan, a glicin i serin su imale pozitivan učinak na razinu ekspresije topljivog proteina. Preciznost linearnog modela predviđanja topljivosti proteina bila je 80 %. Zaključeno je da se povećanjem udjela polarnih aminokiselina, osobito glicina i serina, te smanjenjem udjela arginina bitno povećala ekspresija topljivog proteina HSA dAb.It is widely accepted that features such as pI, length, molecular mass and amino acid (AA) sequence have a significant influence on protein solubility. Here, we mainly focused on AA composition and explored those that most affected the soluble expression level of human serum albumin (HSA) domain antibody (dAb). The soluble expression and sequence of 65 dAb variants were analysed using clustering and linear modelling. Certain AAs significantly affected the soluble expression level of dAb, with the specific AA combinations being (S, R, N, D, Q), (G, R, C, N, S) and (R, S, G); these combinations respectively affected the dAb expression level in the broth supernatant, the level in the pellet lysate and total soluble dAb. Among the 20 AAs, R displayed a negative influence on the soluble expression level, whereas G and S showed positive effects. A linear model was built to predict the soluble expression level from the sequence; this model had a prediction accuracy of 80 %. In summary, increasing the content of polar AAs, especially G and S, and decreasing the content of R, was helpful to improve the soluble expression level of HSA dAb

    Effect of Particle Size on the Wear Property of Magnetorheological Fluid

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    Aiming to study the effect of particle size on the wear property of magnetorheological fluid (MRF), experiment materials, preparation process, and test methods are elaborated, and three different MRF samples consisting of particles of different size are prepared. Test experiments are carried out and the effect of particle size on the wear property of MRF is discussed. Moreover, the microstructures of particles extracted from MRF obtained before and after the wear experiments are observed by scanning electron microscope (SEM). Experimental results show that the particle size has a significant effect on wear property of MRF. Furthermore, the MRF with particles of 1.5–2.8 μm diameter on average is good for the requirement of engineering applications
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