Evaluation of reference genes in mouse preimplantation embryos for gene expression studies using real-time quantitative RT-PCR (RT-qPCR)

BACKGROUND: Real-time quantitative reverse-transcriptase polymerase chain reaction (RT-qPCR) is the most sensitive, and valuable technique for rare mRNA detection. However, the expression profiles of reference genes under different experimental conditions, such as different mouse strains, developmental stage, and culture conditions have been poorly studied. RESULTS: mRNA stability of the actb, gapdh, sdha, ablim, ywhaz, sptbn, h2afz, tgfb1, 18 s and wrnip genes was analyzed. Using the NormFinder program, the most stable genes are as follows: h2afz for the B6D2F-1 and C57BL/6 strains; sptbn for ICR; h2afz for KOSOM and CZB cultures of B6D2F-1 and C57BL/6 strain-derived embryos; wrnip for M16 culture of B6D2F-1 and C57BL/6 strain-derived embryos; ywhaz, tgfb1, 18 s, 18 s, ywhaz, and h2afz for zygote, 2-cell, 4-cell, 8-cell, molular, and blastocyst embryonic stages cultured in KSOM medium, respectively; h2afz, wrnip, wrnip, h2afz, ywhaz, and ablim for zygote, 2-cell, 4-cell, 8-cell, molular, and blastocyst stage embryos cultured in CZB medium, respectively; 18 s, h2afz, h2afz, actb, h2afz, and wrnip for zygote, 2-cell, 4-cell, 8-cell, molular, and blastocyst stage embryos cultured in M16 medium, respectively. CONCLUSIONS: These results demonstrated that candidate reference genes for normalization of target gene expression using RT-qPCR should be selected according to mouse strains, developmental stage, and culture conditions. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/1756-0500-7-675) contains supplementary material, which is available to authorized users

Alpha 1,3-Galactosyltransferase Deficiency in Pigs Increases Sialyltransferase Activities That Potentially Raise Non-Gal Xenoantigenicity

We examined whether deficiency of the GGTA1 gene in pigs altered the expression of several glycosyltransferase genes. Real-time RT-PCR and glycosyltransferase activity showed that 2 sialyltransferases [α2,3-sialyltransferase (α2,3ST) and α2,6-sialyltransferase (α2,6ST)] in the heterozygote GalT KO liver have higher expression levels and activities compared to controls. Enzyme-linked lectin assays indicated that there were also more sialic acid-containing glycoconjugate epitopes in GalT KO livers than in controls. The elevated level of sialic-acid-containing glycoconjugate epitopes was due to the low level of α-Gal in heterozygote GalT KO livers. Furthermore, proteomics analysis showed that heterozygote GalT KO pigs had a higher expression of NAD+-isocitrate dehydrogenase (IDH), which is related to the CMP-N-acetylneuraminic acid hydroxylase (CMAH) enzyme reaction. These findings suggest the deficiency of GGTA1 gene in pigs results in increased production of N-glycolylneuraminic acid (Neu5Gc) due to an increase of α2,6-sialyltransferase and a CMAH cofactor, NAD+-IDH. This indicates that Neu5Gc may be a critical xenoantigen. The deletion of the CMAH gene in the GalT KO background is expected to further prolong xenograft survival

Analysis of the Complex Causes of Death Accidents Due to Mobile Cranes Using a Modified MEPS Method: Focusing on South Korea

The convenience and efficiency of mobile cranes are expanding their applicability in industrial sites, but fatal accidents continue to occur as their use increases. There were 56 cases in South Korea from 2015 to 2019, killing 59 workers. To accurately investigate the cause of a fatal accident, accident investigation reports were used. Since they are used not only as the cause of the accident but also as a result of judicial treatment, only direct causes are mentioned. Thus, indirect causes in this study were separately analyzed to induce a complex cause analysis. The man-made, management, economic, physical, political, and social (MEPS) analysis method, developed by the National Institute of Disaster in South Korea, is a type of root cause analysis (RCA), used to derive the fundamental causes of various types of disasters, mainly social ones. The complex causes of fatal accidents were analyzed by applying a modified MEPS method to mobile cranes. The MEPS method investigated three categories, namely man-made, management, and physical factors, among six categories and a newly established level four, to find the root cause of fatal accidents. The analysis results showed that violations of procedures and regulations were the most frequent causes in the man-made factors. A lack of general and special safety education was the most common cause in the management factor, and the overturning, falling, and jamming of the mobile crane were the most frequent causes in the physical factor

A Study on the Status of Safety and Health for Golf Course Caddies and Improvement of Protective Measures in South Korea

Due to the rapid expansion of the leisure industry, there were about 32,000 golf caddies in South Korea in 2020, an increase of 18.5% compared to 2016. Consequently, they face an increasing industrial accident rate, which is presumed to be the result of exposure to various harmful factors. Through a survey and oral interview of 221 caddies across more than 20 golf courses, health protective measures, protective measures for caddies, preparation for golf cart operation, physical burden, compliance with golf cart safety during games, and golf course responses to emotional labor were investigated in this study to identify safety and health problems of caddies and suggest prevention measures. Preliminary interviews were conducted to confirm golf courses’ safety and health status and participants’ characteristics. Golf caddies’ health and safety were confirmed using frequency analysis, independent sample t-test, one-way analysis of variance (ANOVA), Pearson correlation analysis, and multiple regression analysis. The results showed that caddies’ workplaces were relatively vulnerable to safety and health issues and caddies were exposed to various harmful risk factors. In addition, it was confirmed that golf caddy protective measures, golf cart safety compliance, physical burden, and health protection affect golf courses’ response level to caddies’ emotional labor

Production of Anti-Cancer Agent Using Microbial Biotransformation

Microbial biotransformation is a great model system to produce drugs and biologically active compounds. In this study, we elucidated the fermentation and production of an anti-cancer agent from a microbial process for regiospecific hydroxylation of resveratrol. Among the strains examined, a potent strain showed high regiospecific hydroxylation activity to produce piceatannol. In a 5 L (w/v 3 L) jar fermentation, this wild type Streptomyces sp. in the batch system produced 205 mg of piceatannol (i.e., 60% yields) from 342 mg of resveratrol in 20 h. Using the product, an in vitro anti-cancer study was performed against a human cancer cell line (HeLa). It showed that the biotransformed piceatannol possessed a significant anticancer activity. This result demonstrates that a biotransformation screening method might be of therapeutic interest with respect to the identification of anti-cancer drugs

The Deformation of Polydimethylsiloxane (PDMS) Microfluidic Channels Filled with Embedded Circular Obstacles under Certain Circumstances

Experimental investigations were conducted to determine the influence of polydimethylsiloxane (PDMS) microfluidic channels containing aligned circular obstacles (with diameters of 172 µm and 132 µm) on the flow velocity and pressure drop under steady-state flow conditions. A significant PDMS bulging was observed when the fluid flow initially contacted the obstacles, but this phenomenon decreased in the 1 mm length of the microfluidic channels when the flow reached a steady-state. This implies that a microfluidic device operating with steady-state flows does not provide fully reliable information, even though less PDMS bulging is observed compared to quasi steady-state flow. Numerical analysis of PDMS bulging using ANSYS Workbench showed a relatively good agreement with the measured data. To verify the influence of PDMS bulging on the pressure drop and flow velocity, theoretical analyses were performed and the results were compared with the experimental results. The measured flow velocity and pressure drop data relatively matched well with the classical prediction under certain circumstances. However, discrepancies were generated and became worse as the microfluidic devices were operated under the following conditions: (1) restricted geometry of the microfluidic channels (i.e., shallow channel height, large diameter of obstacles and a short microchannel length); (2) operation in quasi-steady state flow; (3) increasing flow rates; and (4) decreasing amount of curing agent in the PDMS mixture. Therefore, in order to obtain reliable data a microfluidic device must be operated under appropriate conditions

Physical Properties of PDMS (Polydimethylsiloxane) Microfluidic Devices on Fluid Behaviors: Various Diameters and Shapes of Periodically-Embedded Microstructures

Deformable polydimethylsiloxane (PDMS) microfluidic devices embedded with three differently-shaped obstacles (hexagon, square, and triangle) were used to examine the significant challenge to classical fluid dynamics. The significant factors in determining a quasi-steady state value of flow velocity (v)QS and pressure drop per unit length (∆P/∆x)QS were dependent on the characteristic of embedded microstructures as well as the applied flow rates. The deviation from the theoretical considerations due to PDMS bulging investigated by the friction constant and the normalized friction factor revealed that the largest PDMS bulging observed in hexagonal obstacles had the smallest (∆P/∆x)QS ratios, whereas triangle obstacles exhibited the smallest PDMS bulging, but recorded the largest (∆P/∆x)QS ratios. However, the influence of (v)QS ratio on microstructures was not very significant in this study. The results were close to the predicted values even though some discrepancy may be due to the relatively mean bulging and experimental uncertainty. The influence of deformable PDMS microfluidic channels with various shapes of embedded microstructures was compared with the rigid microchannels. The significant deviation from the classical relation (i.e., f~1/Re) was also observed in hexagonal obstacles and strongly dependent on the channel geometry, the degree of PDMS deformation, and the shapes of the embedded microstructures

Case Studies for Dangerous Dust Explosions in South Korea during Recent Years

Despite recent extensive research and technical development to prevent and mitigate dust explosions, processes that produce and handle combustible materials in the form of powders and dusts, either as a main product or as an undesired by-product, have become a constant dust explosion threat as they become more sophisticated and complicated. This study analyzed the characteristics of 53 dust explosions that occurred in South Korea over the last 30 years, and investigated the differences of dust explosions that happened in various countries, such as Japan, the United States, the United Kingdom, and France. In addition, case studies showed the severity of dust explosions occurring in South Korea. Through the special focus on the three most recent years of dust explosions, the causes and processes of the accidents were identified. Analyses of dust explosions in South Korea show that they were mainly caused by organic matter and metal, and, unfortunately, dust explosions occurred repeatedly during grinding, mixing, and injection of powder materials into facilities. No reported accidents occurred during the production processes of wood or paper during the last three years. Taking these characteristics into account, effective ways to prevent or mitigate dust explosions at workplaces where many dust explosions occurred were suggested

In Vitro Studies on a Microfluidic Sensor with Embedded Obstacles Using New Antibacterial Synthetic Compounds (1-TDPPO) Mixed Prop-2-en-1-one with Difluoro Phenyl

This paper describes the use of an analytical microfluidic sensor for accelerating chemo-repellent response and strong anti-bacterial 1-(Thien-2-yl)-3-(2, 6-difluoro phenyl) prop-2-en-1-one (1-TDPPO). The chemically-synthesized antimicrobial agent, which included prop-2-en-1-one and difluoro phenyl groups, was moving through an optically transparent polydimethylsiloxane (PDMS) microfluidic sensor with circular obstacles arranged evenly. The response, growth and distribution of fluorescent labeling Pseudomonas aeruginosa PAO1 against the antimicrobial agent were monitored by confocal laser scanning microscope (CLSM). The microfluidic sensor along with 1-TDPPOin this study exhibits the following advantages: (i) Real-time chemo-repellent responses of cell dynamics; (ii) Rapid eradication of biofilm by embedded obstacles and powerful antibacterial agents, which significantly reduce the response time compared to classical methods; (iii) Minimal consumption of cells and antimicrobial agents; and (iv) Simplifying the process of the normalization of the fluorescence intensity and monitoring of biofilm by captured images and datasets