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

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

Efficacy and safety of combined immunotherapy and stereotactic radiosurgery in NSCLCBM patients and a novel prognostic nomogram: A real-world study

ObjectiveTo explore the effectiveness of combined immunotherapy (IT) and stereotactic radiosurgery (SRS) and address the gap between evidence-based clinical practice and academic knowledge of optimal timing of IT relative to SRS. In addition, to meet the unmet need for an up-to-date prognostic assessment model in the era of IT.MethodsThe data of 86 non-small cell lung cancer brain metastasis (NSCLCBM) patients treated with SRS to 268 brain metastases (BMs) were retrospectively extracted from our hospital database. The Kaplan–Meier analysis was employed for overall survival (OS) and a log-rank test for comparison between groups. Cox proportional hazards regression models were used to identify the significant prognostic factors. The prognostic nomogram was established utilizing the rms package of R software.ResultsIT was found to be associated with improved OS (from BM diagnosis: HR 0.363, 95% CI 0.199 - 0.661, P &lt; 0.001; from SRS: HR 0.472, 95% CI 0.260 - 0.857, P = 0.014). Individuals who received IT in combination with SRS had better OS than those who didn’t (from the day of BM diagnosis: 16.8 vs. 8.4 months, P = 0.006; from the day of SRS: 12 vs. 7 months, P = 0.037). Peri-SRS timing of IT administration was a significant prognostic factor for OS (from BM diagnosis: HR 0.132, 95% CI 0.034 - 0.517, P = 0.004; from SRS: HR 0.14, 95% CI 0.044 - 0.450, P = 0.001). Initiating IT after SRS led to superior OS than concurrent or before (from BM diagnosis: 26.5 vs. 14.1 vs. 7.1 months; from SRS: 21.4 vs. 9.9 vs. 4.1 months, respectively). Additionally, we build a nomogram incorporating IT, cumulative intracranial tumor volume (CITV), and recursive partitioning analysis (RPA), demonstrating a remarkable prognosis prediction performance for SRS-treated NSCLCBM patients.ConclusionPeri-SRS IT is a promising approach in treating NSCLCBM, as improved OS was observed without significantly increasing adverse events. Receipt of IT post-SRS was associated with superior OS than those who received IT concurrently or before. Incorporating IT and CITV into the RPA index could augment its prognosis assessment value for SRS-treated NSCLCBM patients, predominantly in the wild-type

Brassinosteroids Regulate OFP1, a DLT Interacting Protein, to Modulate Plant Architecture and Grain Morphology in Rice

Brassinosteroids (BRs) regulate important agronomic traits in rice, including plant height, leaf angle, and grain size. However, the underlying mechanisms remain not fully understood. We previously showed that GSK2, the central negative regulator of BR signaling, targets DLT, the GRAS family protein, to regulate BR responses. Here, we identified Ovate Family Protein 1 (OFP1) as a DLT interacting protein. OFP1 was ubiquitously expressed and the protein was localized in both cytoplasm and nucleus. Overexpression of OFP1 led to enlarged leaf angles, reduced plant height, and altered grain shape, largely resembled DLT overexpression plants. Genetic analysis showed that the regulation of plant architecture by OFP1 depends on DLT function. In addition, we found OFP1 was greatly induced by BR treatment, and OsBZR1, the critical transcription factor of BR signaling, was physically associated with the OFP1 promoter. Moreover, we showed that gibberellin synthesis was greatly repressed in OFP1 overexpression plants, suggesting OFP1 participates in the inhibition of plant growth by high BR or elevated BR signaling. Furthermore, we revealed that OFP1 directly interacts with GSK2 kinase, and inhibition of the kinase activity significantly promotes OFP1 protein accumulation in plant. Taken together, we identified OFP1 as an additional regulator of BR responses and revealed how BRs promote OFP1 at both transcription and protein levels to modulate plant architecture and grain morphology in rice

Tensor field Graph-Cut for Image Segmentation: A Non-convex Perspective

Image segmentation is a key component of image analysis, which refers to the process of partitioning the image into multiple segments. Graph cut is widely used in image segmentation by constructing a graph that the minimal cut of this graph would lead to partition the corresponding pixels of the different objects. In this paper, we reconstruct the graph cut problem as a special non-convex optimization problem instead of the traditional maximum flow problem. We extend this nonconvex problem to the hypergraph method and combine it with a tensor field based on a directional bilateral filter bank to achieve segmentation in grayscale images. Accordingly, an efficient minimization algorithm is proposed to solve this non-convex problem with global convergence. Furthermore, we have selected the data of BSDS300 and BSDS500 as tests. Experimental results and evaluation index tests further demonstrate the superiority of the proposed method

Retrieval of Consolidated Spatial Memory in the Water Maze Is Correlated with Expression of pCREB and Egr1 in the Hippocampus of Aged Mice

Objective: To study the relationship of the expression of phosphorylated cyclic AMP response element-binding protein (pCREB) and early growth response protein 1 (Egr1) in the hippocampus of aged mice with retrieval of consolidated spatial memory in a water maze. Methods: Twenty-four aged mice were allocated into no training or probe test (naïve), no training but exposed to the same probe test (NTPRT), received training and probe test (PRT), and received training but no probe test (NPRT) groups. Twelve mice were trained in a water maze over 14 days. After the final probe trial on day 15, all mice were anesthetized and the brains were removed. pCREB immunoreactivity (pCREB-ir) and Egr1 immunoreactivity (Egr1-ir) in the hippocampal CA1 and CA3 areas were examined. Results: pCREB-ir and Egr1-ir in the CA1 and CA3 areas of the NPRT and PRT groups were significantly higher than those of the naïve and NTPRT groups, and those in the PRT group were significantly higher than in the NPRT group. In all groups, pCREB-ir was significantly higher in the CA3 area compared to the CA1 area, while Egr1-ir was significantly higher in the CA1 area compared to the CA3 area. Conclusion: Retrieval, as well as formation, of consolidated spatial memory in the water maze is correlated with expression of pCREB and Egr1 in the hippocampus of aged mice

Improvement of the Gut Microbiota In Vivo by a Short-Chain Fatty Acids-Producing Strain Lactococcus garvieae CF11

Gut microbiota has strong connections with health. Regulating and enhancing gut microbiota and increasing the population of beneficial microorganisms constitutes a new approach to increasing the efficiency of health status. Although it has been shown that Lactococcus can adjust gut microbiota and be beneficial for the host, little is known about whether strains of Lactococcus petauri can improve the gut microbiota. This study focused on the influence of Lactococcus petauri CF11 on the gut microbiome composition and the levels of short-chain fatty acids (SCFAs) in vivo in healthy Sprague Dawley rats. The present results showed that strain CF11 was able to induce a higher amount of fecal acetic acid and propionic acid and enhance species richness. Moreover, strain CF11 improved the gut microbiota community structure. In the experimental group, the genera Oscillospira, Coprococcus, and Ruminococcus, which are reported to be able to produce SCFAs, are significantly increased when compared with the control group (p &lt; 0.05). Finally, the functions of genes revealed that 180 pathways were upregulated or downregulated in comparison with the control group. Among them, the top-five clearly enriched pathways regarding metabolism included porphyrin and chlorophyll metabolism; C5-Branched dibasic acid metabolism; valine, leucine, and isoleucine biosynthesis; phenylalanine, tyrosine, and tryptophan biosynthesis; and ascorbate and aldarate metabolism. Our data suggest that the SCFAs-producing strain CF11 is a potential probiotic