Nicotiana benthamiana protein, NbPCIP1, interacting with Potato virus X coat protein plays a role as susceptible factor for viral infection

AbstractThe interactions of viral coat protein (CP) and host factors play an important role in viral replication and/or host defense mechanism. In this study, we constructed Nicotiana benthamiana cDNA library to find host factors interacting with Potato virus X (PVX) CP. Using yeast two-hybrid assay, we screened 3.3×106 independent yeast transformants from N. benthamiana cDNA library and identified six positive clones. One positive clone, named PVX CP-interacting protein 1 (NbPCIP1), is a plant-specific protein with homologue in N. tabacum (GenBank accession no. AB04049). We confirmed the PVX CP–NbPCIP1 interaction using yeast-two hybrid assay in yeast, protein–protein binding assay in vitro, and bimolecular fluorescent complementation assay in planta. Quantitative real-time RT-PCR analysis showed that the mRNA level of NbPCIP1 increased in PVX-infected N. benthamiana plants as compared to that of healthy plants. The green fluorescent protein (sGFP)-fused NbPCIP1 (NbPCIP1-sGFP) was localized in ER or ER-associated granular-like structure of cells. When we co-express NbPCIP1-sGFP and red fluorescent protein (RFP)-fused PVX CP (PVX CP-RFP), which were introduced by transiently expressing these proteins in N. benthamiana protoplasts and epidermal cells, however, we observed the co-localization of these proteins in the inclusion body-like complex in areas surrounding nucleus. Transient over-expression and transgene silencing of NbPCIP1 assay analysis indicated that NbPCIP1 plays a critical role in viral replication during PVX infection in host plant

Cardiogenic shock

Cardiogenic shock is an acute circulatory failure due to compromised myocardial contractility associated with congenital heart diseases and cardiomyopathies, such as myocarditis. In this article, the authors present a 3-step overview of cardiogenic shock diagnosis and management to restore tissue oxygen delivery. The first step is early recognition of nonspecific signs of the shock. The second step is medical management, monitoring, and repeated assessment. In addition to conventional parameters, biomarkers may be useful to monitor the shock. The final step is mechanical circulatory support, such as ventricular assist devices, for children with the refractory shock. We also briefly describe the shock in multisystem inflammatory syndrome in children with coronavirus disease 2019

Comparison of volume-controlled and pressure-controlled ventilation using a laryngeal mask airway during gynecological laparoscopy

Background: Several publications have reported the successful, safe use of Laryngeal Mask Airway (LMA)-Classic devices in patients undergoing laparoscopic surgery. However, there have been no studies that have examined the application of volume-controlled ventilation (VCV) or pressure-controlled ventilation (PCV) using a LMA during gynecological laparoscopy. The aim of this study is to compare how the VCV and PCV modes and using a LMA affect the pulmonary mechanics, the gas exchange and the cardiovascular responses in patients who are undergoing gynecological laparoscopy. Methods: Sixty female patients were randomly allocated to one of two groups, (the VCV or PCV groups). In the VCV group, baseline ventilation of the lung was performed with volume-controlled ventilation and a tidal volume of 10 ml/kg ideal body weight (IBW). In the PCV group, baseline ventilation of the lung using pressure-controlled ventilation was initiated with a peak airway pressure that provided a tidal volume of 10 ml/kg IBW and an upper limit of 35 cmH2O. The end-tidal CO2, the peak airway pressures (Ppeak), the compliance, the airway resistance and the arterial oxygen saturation were recorded at T1: 5 minutes after insertion of the laryngeal airway, and at T2 and T3: 5 and 15 minutes, respectively, after CO2 insufflation. Results: The Ppeak at 5 minutes and 15 minutes after CO2 insufflation were significantly increased compared to the baseline values in both groups. Also, at 5 minutes and 15 minutes after CO2 insufflation, there were significant differences of the Ppeak between the two groups. The compliance decreased in both groups after creating the pneumopertoneim (P < 0.05). Conclusions: Our results demonstrate that PCV may be an effective method of ventilation during gynecological laparoscopy, and it ensures oxygenation while minimizing the increases of the peak airway pressure after CO2 insufflation. ��� the Korean Society of Anesthesiologists, 2011

Prediction of novel synthetic pathways for the production of desired chemicals

<p>Abstract</p> <p>Background</p> <p>There have been several methods developed for the prediction of synthetic metabolic pathways leading to the production of desired chemicals. In these approaches, novel pathways were predicted based on chemical structure changes, enzymatic information, and/or reaction mechanisms, but the approaches generating a huge number of predicted results are difficult to be applied to real experiments. Also, some of these methods focus on specific pathways, and thus are limited to expansion to the whole metabolism.</p> <p>Results</p> <p>In the present study, we propose a system framework employing a retrosynthesis model with a prioritization scoring algorithm. This new strategy allows deducing the novel promising pathways for the synthesis of a desired chemical together with information on enzymes involved based on structural changes and reaction mechanisms present in the system database. The prioritization scoring algorithm employing Tanimoto coefficient and group contribution method allows examination of structurally qualified pathways to recognize which pathway is more appropriate. In addition, new concepts of binding site covalence, estimation of pathway distance and organism specificity were taken into account to identify the best synthetic pathway. Parameters of these factors can be evolutionarily optimized when a newly proven synthetic pathway is registered. As the proofs of concept, the novel synthetic pathways for the production of isobutanol, 3-hydroxypropionate, and butyryl-CoA were predicted. The prediction shows a high reliability, in which experimentally verified synthetic pathways were listed within the top 0.089% of the identified pathway candidates.</p> <p>Conclusions</p> <p>It is expected that the system framework developed in this study would be useful for the <it>in silico </it>design of novel metabolic pathways to be employed for the efficient production of chemicals, fuels and materials.</p

Application and evaluation of the MLVA typing assay for the Brucella abortus strains isolated in Korea

<p>Abstract</p> <p>Background</p> <p>A Brucella eradication program has been executed in Korea. To effectively prevent and control brucellosis, a molecular method for genetic identification and epidemiological trace-back must be established. As part of that, the MLVA typing assay was evaluated and applied to <it>B. abortus </it>isolates for analyzing the characteristics of the regional distribution and relationships of foreign isolates.</p> <p>Results</p> <p>A total of 177 isolates originating from 105 cattle farms for the period 1996 to 2008 were selected as representatives for the nine provinces of South Korea. A dendrogram of strain relatedness was constructed in accordance with the number of tandem repeat units for 17 loci so that it was possible to trace back in the restricted areas. Even in a farm contaminated by one source, however, the <it>Brucella </it>isolates showed an increase or decrease in one TRs copy number at some loci with high DI values. Moreover, those 17 loci was confirmed in stability via <it>in-vitro </it>and <it>in-vivo </it>passage, and found to be sufficiently stable markers that can readily identify the inoculated strain even if minor changes were detected. In the parsimony analysis with foreign <it>Brucella </it>isolates, domestic isolates were clustered distinctively, and located near the Central and Southern American isolates.</p> <p>Conclusion</p> <p>The MLVA assay has enough discrimination power in the <it>Brucella </it>species level and can be utilized as a tool for the epidemiological trace-back of the <it>B. abortus </it>isolates. But it is important to consider that <it>Brucella </it>isolates may be capable of undergoing minor changes at some loci in the course of infection or in accordance with the changes of the host.</p

Highly sensitive colorimetric detection of allergies based on an immunoassay using peroxidase-mimicking nanozymes

Nanomaterials that exhibit enzyme-like characteristics, which are called nanozymes, have recently attracted significant attention due to their potential to overcome the intrinsic limitations of natural enzymes, such as low stability and relatively high cost for preparation and purification. In this study, we report a highly efficient colorimetric allergy detection system based on an immunoassay utilizing the peroxidase- mimicking activity of hierarchically structured platinum nanoparticles (H-Pt NPs). The H-Pt NPs had a diameter of 30 nm, and were synthesized by a seed-mediated growth method, which led to a significant amount of peroxidase-like activity. This activity mainly occurs because of the high catalytic power of the Pt element, and the fact that the H-Pt NPs have a large surface area available for catalytic events. The H-Pt NPs were conjugated to an antibody for the detection of immunoglobulin E (IgE) in the analytes; IgE is a representative marker for the diagnosis of allergies. They were then successfully integrated into a conventionally used allergy diagnostic test, the ImmunoCAP diagnostic test, as a replacement for natural signaling enzymes. Using this strategy, total and specific IgE levels were detected within 5 min at room temperature, with high specificity and sensitivity. The practical utility of the immunoassay was also successfully verified by correctly determining the levels of both total and specific IgE in real human serum samples with high precision and reproducibility. The present H-Pt NP-based immunoassay system would serve as a platform for rapid, robust, and convenient analysis of IgE, and can be extended to the construction of diagnostic systems for a variety of clinically important target molecules.11Ysciescopu

Antioxidant effect of lidocaine and procaine on reactive oxygen species-induced endothelial dysfunction in the rabbit abdominal aorta

Background: Reactive oxygen species (ROS) induce lipid peroxidation and tissue damage in the endothelium. We tested the antioxidant effect of lidocaine and procaine on ROS-induced endothelial damage in the rabbit aorta. Methods: Aortic rings isolated from rabbits were suspended in an organ bath filled with Krebs-Henseleit (K-H) solution bubbled with 5% CO2 and 95% O 2 at 37.5??C. After precontraction with phenylephrine (PE, 10 -6 M), changes in tension were recorded following a cumulative administration of acetylcholine (ACh 3 ?? 10-8 to 10 -6 M). Differences were measured as percentages of ACh-induced relaxation of aortic rings before and after exposure to ROS as generated by electrolysis of the K-H solution. The aortic rings were pretreated with lidocaine or procaine (10-5 M to 3 ?? 10-3 M) to compare their effects, as well as ROS scavengers, catalase, mannitol, sodium salicylate, and deferoxamine, and a catalase inhibitor, 3-amino-1,2,4-triazole (3AT). Results: Lidocaine and procaine dose-dependently maintained endothelium-dependent relaxation induced by ACh despite ROS activity (P &lt; 0.05 vs control value). The 3AT pretreated procaine (3 ?? 10-3 M) group decreased more significantly than the un-pretreated procaine group (P &lt; 0.05). Conclusions: These findings suggest that lidocaine and procaine dose-dependently preserve endothelium-dependent vasorelaxation against ROS attack, potentially via hydrogen peroxide scavenging. Copyright ?? Korean Society of Anesthesiologists, 2010