Proteomic analysis of the endophytic fungus Undifilum oxytropis

The filamentous Ascomycete fungus Undifilum oxytropis is an endophyte of locoweed plants of the genera Oxytropis that produces a toxic alkaloid swainsonine. Swainsonine, an alpha-mannosidase inhibitor causes a general toxicosis and neurological problems (locoism) when consumed by grazing animals. Swainsonine is also being assessed for its anti-cancer properties. While the ecology of U. oxytropis has been studied, little is known about the genetics and proteomics of any swainsonine-producing fungus. To help understand the proteins in U. oxytropis, the proteome of U. oxytropis was analyzed using 2-dimensional electrophoresis (2-DE). Proteins from U. oxytropis mycelia were extracted and separated by in-gel isoelectric focusing (IEF). The entire immobilized pH gradient (IPG) strip was cut into a set of gel sections and each gel section was digested with trypsin and then identified using liquid chromatography tandem mass spectrometry (LC-MS/MS). 2-DE maps were also developed for U. oxytropis to define its proteome. In the isoelectric point (pI) range of 3-11 and 10-250 kDa ranges, more than 450 spots were detected in 2-DE silver-stained gels, and 52 proteins were identified by LC-MS/MS. Most of the identified proteins were involved in energy production, oxidoreductase activity, carbohydrate metabolic process, amino acid and cellular ketone metabolic process. A large group of identified proteins were related to stress proteins and heat shock proteins. This work presents the first two-dimensional reference map of this alkaloid-producing fungus. Details of the proteome serve as a baseline for further study of this swainsonine-producing fungus and are essential for a reverse genetic analysis of the fungus.Keywords: Undifilum oxytropis fungus, two-dimensional gel electrophoresis, proteome reference map, liquid chromatography tandem mass spectrometry, swainsonin

Relationship between quality of professional capacity building for kindergarten teachers and children’s language development: the mediating role of kindergarten language education activities quality

Strengthening the professional capacity building for kindergarten teachers is crucial for improving the quality of kindergarten education and promoting children’s development. In order to explore the impact of the quality of professional capacity building for kindergarten teachers and kindergarten language education activities quality on children’s language development, a study was conducted. A total of 1,584 children from 90 kindergartens in 5 provinces in China were randomly selected as research participants. Utilizing an independent sample t-test and correlation analysis (with SPSS 26.0 as the tool), the study analyzed the disparities in the professional capacity building for kindergarten teachers of different types, kindergarten language education activities quality, and the children’s language developmental levels, as well as the interrelationships among these factors. A multilevel structural equation model (using MPLUS 8.3 as the tool) was employed to analyze the mediating role that the kindergarten language education activities quality plays between the quality of professional capacity building for kindergarten teachers and children’s language development levels. The results indicate that children in public kindergartens demonstrate significantly higher language development levels than those in private kindergartens. A positive correlation exists between the quality of professional capacity building for kindergarten teachers, kindergarten language education activities quality and children’s language development levels. Furthermore, kindergarten language education activities quality shows partial mediating effects on the relationship between them. These findings highlight the need to enhance the quality of education in private kindergartens, reduce the disparities in language development levels between private and public kindergartens, and reinforce the quality of professional capacity building for kindergarten teachers to provide effective support for children’s language development. Ultimately, the study suggests concentrating on the organization and implementation of kindergarten language education activities as a means of evaluating the quality of professional capacity building for kindergarten teachers

Adaptive Quantization for Predicting Transform-based Point Cloud Compression

The representation of three-dimensional objects with point clouds is attracting increasing interest from researchers and practitioners. Since this representation requires a huge data volume, effective point cloud compression techniques are required. One of the most powerful solutions is the Moving Picture Experts Group geometry-based point cloud compression (G-PCC) emerging standard. In the G-PCC lifting transform coding technique, an adaptive quantization method is used to improve the coding efficiency. Instead of assigning the same quantization step size to all points, the quantization step size is in-creased according to level of detail traversal order. In this way, the attributes of more important points receive a finer quantization and have a smaller quantization error than the attributes of less important ones. In this paper, we adapt this approach to the G-PCC predicting transform and propose a hardware-friendly weighting method for the adaptive quantization. Experimental results show that compared to the current G-PCC test model, the proposed method can achieve an average Bjøntegaard delta rate of -6.7%, -14.7%, -15.4%, and -10.0% for the luma, chroma Cb, chroma Cr, and reflectance components, respectively on the MPEG Cat1-A, Cat1-B, Cat3-fused and Cat3-frame datasets

Investigating long-chain polyunsaturated fatty acid biosynthesis in teleost fish: Functional characterization of fatty acyl desaturase (Fads2) and Elovl5 elongase in the catadromous species, Japanese eel Anguilla japonica

The capacity for endogenous production of LC-PUFA from PUFA in euryhaline or diadromous fish is largely unknown other than for Atlantic salmon (Salmo salar), an anadromous species, which displays a freshwater pattern. The aim of the present study was to characterize the enzymes of the LC-PUFA pathway in Japanese eel (Anguilla japonica), the most important catadromous species currently being farmed. cDNAs of two key genes were cloned and functional assays showed they encoded a desaturase (Fads2) with D6 and D8 activity and an elongase (Elovl5) with activity towards C18 and C20 PUFA, with activities similar to marine fish and an D6/D8 activity ratio similar to Atlantic salmon. Furthermore, tissue distribution of the mRNA showed a clear marine pattern with highest expression in brain and eye. Phylogenetic analysis placed the eel cDNAs in line with classical taxonomy. The data suggest that diadromous species display a pattern of LC-PUFA biosynthesis capacity that likely reflects the environmental and nutritional influence of their early life stages rather than those of adult fish. Future studies aim to establish the full range of PUFA desaturases and elongases in Japanese eel and to provide further insight to the importance and relevance of LC-PUFA biosynthesis in fish species and the influence of diadromy

Convergence of regional economic cycles in Turkey

Dissimilar economic fluctuations and asymmetric shocks across the regions of a country might create severe policy distortions that, under these circumstances, aggregate policy interventions (such as taxation and interest rates), are likely to be sub-optimal for at least a fraction of the regions. For instance, monetary policy can hardly satisfy the needs of all regions when some of the regions are experiencing a boom while others are in a recession phase. For these reasons, similarity of regional business cycles and their convergence are highly desirable from a policy viewpoint. The aim of this paper is, therefore, to provide empirical evidence and policy implications in that context. In particular, I analyze business cycle correlations across Turkish provinces and the tendency of these cycles to converge over the period of analysis between 1975-2000 and 2004-2008 (for Nomenclature of Territorial Units for Statistics [NUTS]-2 regions). I find that regional business cycle asymmetries have tended to decrease in recent decades. This result, although it seems to provide evidence in favor of rising correlations, shows that the convergence process is rather slow and there still exist asymmetries across the regional business cycles

Detection of Brucellae in peripheral blood mononuclear cells for monitoring therapeutic efficacy of brucellosis infection.

BACKGROUND: Brucellosis is one of the most severe widespread zoonoses caused by the Gram-negative bacterium Brucella species. The diagnosis and clinical assessment of human brucellosis are very important for the management of patients, while there is a lack of effective methods to detect Brucellae. Classical culture of Brucella species is time consuming and often fails. A simple and sensitive assay is needed for diagnosis of Brucella infection and monitoring of treatment in man. METHODS: Blood samples and peripheral blood mononuclear cells (PBMCs) were collected from 154 patients hospitalized for brucellosis. Brucella antibodies were detected by Rose Bengal Plate Test (RBPT), Standard Tube Agglutination Test (SAT) and enzyme-linked immunosorbent assay (ELISA). Intracellular Brucellae were detected by blood culture and immunofluorescence staining (IFS). RESULTS: Among 154 brucellosis patients, 59.7% (92/154) were antibody reactive by RBPT, 81.8% (126/154) by SAT and 95.5% (147/154) by ELISA, respectively. Only 3.2% (5/154) of patient blood samples resulted in positive Brucella culture, while 68.8% (106/154) carried IFS detectable Brucella antigens in PBMCs. Gender (P = 0.01) but not age (P > 0.05) was a significant risk factor. The frequency of intracellular Brucella antigens was similar between patients receiving different treatment regimens (P > 0.05). However, a significant decrease of intracellular Brucellae was observed only in patients with acute brucellosis after the third course of treatment (P < 0.05), suggesting that current regimens to treat chronic brucellosis were not effective. CONCLUSIONS: IFS appears a sensitive assay for detection of Brucella antigens in PBMCs and could be used for diagnosis and therapeutic monitoring of brucellosis in clinical practice

Evaluation of Reactivity of Monoclonal Antibodies Against Omp25 of Brucella spp.

Brucellosis is a serious zoonosis occurring mainly in developing countries, and its diagnosis is largely dependent on serologic detection and bacterial culture. In this study, we developed the murine monoclonal antibodies (mAbs) against a conserved and major outer membrane protein 25 (Omp25) of Brucella species (B. spp.) for use in clinical diagnosis. The mAbs to Omp25 were produced by hybridoma technique, which were utilized for developing various immunoassays for detection of Brucellae, including Western blot (WB), enzyme-linked immunosorbent assay (ELISA), immunochemical staining (ICS), immunofluorescence staining (IFS), and flow cytometry assay (FCM). A number of five mAbs (2B10, 4A12, 4F10, 6C12, and 8F3) specific to Omp25 were selected, including 2 IgG1, 2 IgG2a, and 1 IgG2b. Among them, mAbs 6C12, 8F3, and 4A12 reacted highly with B. melitensis (M5-90), B. abortus (S19, 104M, and 2308), and B. suis strain (S2). No cross-reactivity with Yersinia enterocolitica O:9, Salmonella spp., and Escherichia coli was found. By mapping Omp25 epitopes, mAb 6C12 was found as reacting with a semi-conformational epitope, and mAbs 4A12 and 8F3 as recognizing a different linear epitope, respectively. The paired mAbs were tested for detecting Brucella species, suggesting that 8F3 was suitable for solid phase capture and 6C12 or 4A12 was suitable for conjugation with HRP for detection of Brucella Omp25 in ELISA. The FCM was established by mAb 6C12 for detecting intracellular Brucellae-infected peripheral blood mononuclear cells (PBMCs) from brucellosis patients. In conclusion, mAbs against Omp25 are precious reagents for detection of Brucellae in clinical samples with various immunoassays. mAb 6C12-based FCM could be potentially used for the monitoring of therapeutic efficacy for brucellosis in clinical practice

Location of modulatory β subunits in BK potassium channels

Large-conductance voltage- and calcium-activated potassium (BK) channels contain four pore-forming α subunits and four modulatory β subunits. From the extents of disulfide cross-linking in channels on the cell surface between cysteine (Cys) substituted for residues in the first turns in the membrane of the S0 transmembrane (TM) helix, unique to BK α, and of the voltage-sensing domain TM helices S1–S4, we infer that S0 is next to S3 and S4, but not to S1 and S2. Furthermore, of the two β1 TM helices, TM2 is next to S0, and TM1 is next to TM2. Coexpression of α with two substituted Cys’s, one in S0 and one in S2, and β1 also with two substituted Cys’s, one in TM1 and one in TM2, resulted in two αs cross-linked by one β. Thus, each β lies between and can interact with the voltage-sensing domains of two adjacent α subunits