A New Reporter Vector System Based on Flow-Cytometry to Detect Promoter Activity

In this study, we report the development of a new dual reporter vector system for the analysis of promoter activity. This system employs green fluorescence emitting protein, EGFP, as a reporter, and uses red fluorescence emitting protein, DsRed, as a transfection control in a single vector. The expression of those two proteins can be readily detected via flow cytometry in a single analysis, with no need for any further manipulation after transfection. As this system allows for the simultaneous detection of both the control and reporter proteins in the same cells, only transfected cells which express the control protein, DsRed, can be subjected to promoter activity analysis, via the gating out of all un-transfected cells. This results in a dramatic increase in the promoter activity detection sensitivity. This novel reporter vector system should prove to be a simple and efficient method for the analysis of promoter activity

Modulatory role of phospholipase D in the activation of signal transducer and activator of transcription (STAT)-3 by thyroid oncogenic kinase RET/PTC

<p>Abstract</p> <p>Background</p> <p>RET/PTC (rearranged in transformation/papillary thyroid carcinomas) gene rearrangements are the most frequent genetic alterations identified in papillary thyroid carcinoma. Although it has been established that RET/PTC kinase plays a crucial role in intracellular signaling pathways that regulate cellular transformation, growth, and proliferation in thyroid epithelial cells, the upstream signaling that leads to the activation of RET/PTC is largely unknown. Based on the observation of high levels of PLD expression in human papillary thyroid cancer tissues, we investigated whether PLD plays a role in the regulating the RET/PTC-induced STAT3 activation.</p> <p>Methods</p> <p>Cancer tissue samples were obtained from papillary thyroid cancer patients (n = 6). The expression level of PLD was examined using immunohistochemistry and western blotting. Direct interaction between RET/PTC and PLD was analyzed by co-immunoprecipitation assay. PLD activity was assessed by measuring the formation of [³H]phosphatidylbutanol, the product of PLD-mediated transphosphatidylation, in the presence of <it>n</it>-butanol. The transcriptional activity of STAT3 was assessed by m67 luciferase reporter assay.</p> <p>Results</p> <p>In human papillary thyroid cancer, the expression levels of PLD2 protein were higher than those in the corresponding paired normal tissues. PLD and RET/PTC could be co-immunoprecipitated from cells where each protein was over-expressed. In addition, the activation of PLD by pervanadate triggered phosphorylation of tyrosine 705 residue on STAT-3, and its phosphorylation was dramatically higher in TPC-1 cells (from papillary carcinoma) that have an endogenous RET/PTC1 than in ARO cells (from anaplastic carcinoma) without alteration of total STAT-3 expression. Moreover, the RET/PTC-mediated transcriptional activation of STAT-3 was synergistically increased by over-expression of PLD, whereas the PLD activity as a lipid hydrolyzing enzyme was not affected by RET/PTC.</p> <p>Conclusion</p> <p>These findings led us to suggest that the PLD synergistically functions to activate the STAT3 signaling by interacting directly with the thyroid oncogenic kinase RET/PTC.</p

A Study on Technology Development Performance and Technology Commercialization Performance According to the Technology Development Capability of SMEs Focusing on a Comparative Analysis of Technology Business Groups

Recently, a lot of small- and mid-sized companies have emerged through continuous technological development and start-up successes despite insufficient scales and resources compared to medium companies or conglomerates in fierce market competition. Technological development capability, which is required to acquire, select, or utilize source technology for company competitiveness, becomes the competitive edge and key capability to gain distinctiveness. Accordingly, the objective of this research was to investigate the influential factors on performance by suggesting technological development results and technology commercialization as the outcome of technological development capability and to identify the capability required for successful technological development performance and technology commercialization for companies through previous academic research. Moreover, the study aimed to investigate performance differences according to technology business groups by dividing the groups into high-technology companies, medium-technology companies, and low-technology companies. As a result of the analysis, among the indicators constituting technological development capability, manpower in technological development and technological development-specialized research centers had a positive influence on the product competitiveness of technological development performance. Also, technological development expenses and technology skill levels were verified to have positive impacts on the technology competitiveness of technological development performance. As for technology commercialization performance, the more manpower that is in research and development, the higher the sales amounts by technological development; also, the higher the technology skill levels, the higher the export amounts by technological development. Moreover, technology business groups had differences in performance. This research has significance in focusing on the influence of the technological development capability of companies on technological development performance and technology commercialization performance. Future research is expected to draw new variations different from the analysis results from existing studies by setting variations, investigating complementary relationships among variations in detail, and utilizing statistical techniques that can control mutual relationships among variations

Immunogenicity after Second ChAdOx1 nCoV-19 (AZD1222) Vaccination According to the Individual Reactogenicity, Health Status and Lifestyle

The immune-acquired responses after vaccination vary depending on the type of vaccine and the individual. The purpose of this study was to investigate the relationship between the acquisition of immunity and the side effects, health status, and lifestyle after completion of the second dose of AZD1222. Blood samples were collected after a second dose of AZD1222. The Euroimmun Anti-SARS-CoV-2 ELISA (IgG) for anti-S1 antibody, the cPASS SARS-CoV-2 neutralizing antibody detection kit for the surrogate virus neutralization test, and the T-spot Discovery SARS-CoV-2 kit were used to identify cellular immunogenicity. Patient experience of adverse effects was investigated using questionnaires. Information on health status and lifestyle were collected from the most recent health checkup data. Generally, females experience more reactogenicity in both intensity and duration. The rash of the first shot and chills of the second shot were associated with humoral immunity. However, comprehensive adverse effects had no correlation with humoral and cellular immunity. The T-spot-positive group had a higher creatinine level, which reflects muscle mass, than the T-spot-negative group. Males presented a higher level of T-spot assays. Body mass index and age were negatively correlated with the T-spot assay and anti-S1 antibody, respectively. Immune acquisition after the second AZD1222 shot was not associated with reactogenicity. However, individuals&rsquo; sex, age, and BMI were found to be associated with immunogenicity after vaccination

Could Natural Products Help in the Control of Obesity? Current Insights and Future Perspectives

Obesity is a global issue faced by many individuals worldwide. However, no drug has a pronounced effect with few side effects. Green tea, a well-known natural product, shows preventive effects against obesity by decreasing lipogenesis and increasing fat oxidation and antioxidant capacity. In contrast, other natural products are known to contribute to obesity. Relevant articles published on the therapeutic effect of natural products on obesity were retrieved from PubMed, Web of Science, and Scopus. The search was conducted by entering keywords such as “obesity”, “natural product”, and “clinical trial”. The natural products were classified as single compounds, foods, teas, fruits, herbal medicines—single extract, herbal medicines—decoction, and herbal medicines—external preparation. Then, the mechanisms of these medicines were organized into lipid metabolism, anti-inflammation, antioxidation, appetite loss, and thermogenesis. This review aimed to assess the efficacy and mechanisms of effective natural products in managing obesity. Several clinical studies reported that natural products showed antiobesity effects, including Coffea arabica (coffee), Camellia sinensis (green tea), Caulerpa racemosa (green algae), Allium sativum (garlic), combined Ephedra intermedia Schrenk, Thea sinensis L., and Atractylodes lancea DC extract (known as Gambisan), Ephedra sinica Stapf, Angelica Gigantis Radix, Atractylodis Rhizoma Alba, Coicis semen, Cinnamomi cortex, Paeoniae radix alba, and Glycyrrhiza uralensis (known as Euiiyin-tang formula). Further studies are expected to refine the pharmacological effects of natural products for clinical use

Taxonomic and Biochemical Characterization of Microalga Graesiella emersonii GEGS21 for Its Potential to Become Feedstock for Biofuels and Bioproducts

Graesiella emersonii is a commercially exploitable source of bioactive compounds and biofuels with potential applications in microalgae-based industries. Despite this, little taxonomical information is available. Therefore, proper identification and characterization are needed for the sustainable utilization of isolated microalgae. In this study, an axenically isolated unicellular green alga from the Geumgang Estuary, Korea was investigated for its morphological, molecular, and biochemical characteristics. The morphological characteristics were typical of G. emersonii. Molecular phylogenetic analysis of the 18S rDNA sequence verified that the isolate belonged to G. emersonii and was subsequently named G. emersonii GEGS21. It was isolated from brackish water, and its optimal growth temperature, salinity, and light intensity were at 28&ndash;32 &deg;C, 0 M NaCl, and 130&ndash;160 &micro;mol m&minus;2 s&minus;1, respectively. The strain thrived over a range of temperatures (5&ndash;40 &deg;C) and withstood up to 0.5 M NaCl. The isolate was rich in omega-6 linoleic acid (C18:2 n-6, 26.3%) and palmitic acid (C16:0, 27.5%). The fuel quality properties were determined, and biodiesel from GEGS21 could be used as a biodiesel blend. Value-added carotenoids lutein (1.5 mg g&minus;1 dry cell weight, DCW) and neoxanthin (1.2 mg g&minus;1 DCW) were biosynthesized as accessory pigments by this microalga. The biomass of this microalga may serve as feedstock for biodiesel production as well as producing valuable &omega;-6 and carotenoids

Taxonomic and Biochemical Characterization of Microalga <i>Graesiella emersonii</i> GEGS21 for Its Potential to Become Feedstock for Biofuels and Bioproducts

Graesiella emersonii is a commercially exploitable source of bioactive compounds and biofuels with potential applications in microalgae-based industries. Despite this, little taxonomical information is available. Therefore, proper identification and characterization are needed for the sustainable utilization of isolated microalgae. In this study, an axenically isolated unicellular green alga from the Geumgang Estuary, Korea was investigated for its morphological, molecular, and biochemical characteristics. The morphological characteristics were typical of G. emersonii. Molecular phylogenetic analysis of the 18S rDNA sequence verified that the isolate belonged to G. emersonii and was subsequently named G. emersonii GEGS21. It was isolated from brackish water, and its optimal growth temperature, salinity, and light intensity were at 28–32 °C, 0 M NaCl, and 130–160 µmol m−2 s−1, respectively. The strain thrived over a range of temperatures (5–40 °C) and withstood up to 0.5 M NaCl. The isolate was rich in omega-6 linoleic acid (C18:2 n-6, 26.3%) and palmitic acid (C16:0, 27.5%). The fuel quality properties were determined, and biodiesel from GEGS21 could be used as a biodiesel blend. Value-added carotenoids lutein (1.5 mg g−1 dry cell weight, DCW) and neoxanthin (1.2 mg g−1 DCW) were biosynthesized as accessory pigments by this microalga. The biomass of this microalga may serve as feedstock for biodiesel production as well as producing valuable ω-6 and carotenoids

RNA demethylation by FTO stabilizes the FOXJ1 mRNA for proper motile ciliogenesis

Adenosine N6-methylation (m6A) is one of the most pervasive mRNA modifications, and yet the physiological significance of m6A removal (demethylation) remains elusive. Here, we report that the m6A demethylase FTO functions as a conserved regulator of motile ciliogenesis. Mechanistically, FTO demethylates and thereby stabilizes the mRNA that encodes the master ciliary transcription factor FOXJ1. Depletion of Fto in Xenopus laevis embryos caused widespread motile cilia defects, and Foxj1 was identified as one of the major phenocritical targets. In primary human airway epithelium, FTO depletion also led to FOXJ1 mRNA destabilization and a severe loss of ciliated cells with an increase of neighboring goblet cells. Consistently, Fto knockout mice showed strong asthma-like phenotypes upon allergen challenge, a result owing to defective ciliated cells in the airway epithelium. Altogether, our study reveals a conserved role of the FTO-FOXJ1 axis in embryonic and homeostatic motile ciliogenesis.