Real-Time Temporal Dynamics of Bicistronic Expression Mediated by Internal Ribosome Entry Site and 2A Cleaving Sequence

Multicistronic elements, such as the internal ribosome entry site (IRES) and 2A-like cleavage sequence, serve crucial roles in the eukaryotic ectopic expression of exogenous genes. For utilization of multicistronic elements, the cleavage efficiency and order of elements in multicistronic vectors have been investigated; however, the dynamics of multicistronic element-mediated expression remains unclear. Here, we investigated the dynamics of encephalomyocarditis virus (EMCV) IRES- and porcine teschovirus-1 2A (p2A)-mediated expression. By utilizing real-time fluorescent imaging at a minute-level resolution, we monitored the expression of fluorescent reporters bridged by either EMCV IRES or p2A in two independent cultured cell lines, HEK293 and Neuro2a. We observed significant correlations for the two fluorescent reporters in both multicistronic elements, with a higher correlation coefficient for p2A in HEK293 but similar coefficients for IRES-mediated expression and p2A-mediated expression in Neuro2a. We further analyzed the causal relationship of multicistronic elements by convergent cross mapping (CCM). CCM revealed that in all four conditions examined, the expression of the preceding gene causally affected the dynamics of the subsequent gene. As with the cross correlation, the predictive skill of p2A was higher than that of IRES in HEK293, while the predictive skills of the two multicistronic elements were indistinguishable in Neuro2a. To summarize, we report a significant temporal correlation in both EMCV IRES- and p2A-mediated expression based on the simple bicistronic vector and real-time fluorescent monitoring. The current system also provides a valuable platform to examine the dynamic aspects of expression mediated by diverse multicistronic elements under various physiological conditions.National Research Foundation of Korea (NRF) of the Ministry of Science and ICT [NRF-2017R1C1B2008775, NRF-2017R1A4A1015534, NRF-2018M3C7A1022310]; KBRI basic research program through Korea Brain Research Institute - Ministry of Science and ICT [17-BR-04]Open access journalThis item from the UA Faculty Publications collection is made available by the University of Arizona with support from the University of Arizona Libraries. If you have questions, please contact us at [email protected]

Measurement of epigenetic alterations from patient’s tissues in myoma, adenomyoma and endometriosis

Background: Myoma, adenomyoma, and endometriosis are estrogen-dependent gynecologic diseases and result in reproductive dysfunction and pelvic pain in women. However, these gynecologic diseases have a complex and poorly understood etiology, involving both genetic and environmental factors. Epigenetic alterations, heritable changes that can modify gene expression without affecting genetic sequence, are associated with the development and progression of numerous pathological states and diseases. Therefore, there is great potential for the use of epigenetics as biomarkers to better understand the early-stage biological responses and molecular mechanisms of gynecologic diesases. We aimed to examine levels of global DNA and gene-specific methylation, which are epigenetic alterations that could be associated with development of gynecologic diseases, including myoma, adenomyoma, and endometriosis. Methods: We measured global DNA methylation (LINE-1) as well as disease relevant gene-specific methylation (i.e. ER, PR, and aromatase) using pyrosequencing assay. For this measurement, gene-specific primers for the selected genes were designed using the Pyro-Mark assay design software. Genomic DNAs from each tissue were extracted, and underwent bisulfite modification to convert unmethylated cytosine residues to uracil. A Pyromark Q96 MD was used for all subsequent pyrosequencing. Samples were processed in duplicates on plates with water controls. Percent methylation of a sample was calculated by averaging all of the interrogated CpG sites. Results: Different methylation levels of selected genes were measured from myoma, adenomyoma, and endometriosis tissues. Our obtained results suggest that epigenetic changes are involved in development of different types of gynecologic diseases

A Study on Apparel Design for Chiropractic Training

In severe cases of spinal misalignment due to wrong back posture during adolescence or focusing on work, modern people can have back pain. Such a symptom is commonly referred to as scoliosis and it already became one of the lifestyle diseases. Hence, the purpose of the present study was to develop a functional musculoskeletal correction wear that can help with maintaining a correct posture by relaxing tonic muscles and maximizing the tension of the relaxed muscle using the principle of sports taping. Market research for design development was conducted at department stores and specialty road shops separately. Department stores were visited on weekdays when the store is not crowded, and specialty road shops were visited at around three o’clock on weekdays after checking their opening, lunch, and closing times. Since the apparel should not interfere with exercise movements while maintaining the body shape, the survey was carried out mostly on yoga and lifestyle training apparel focusing on elasticity and percentage of blending fiber. A camera and audio recorder were prepared and took pictures of clothing tags that show the ratio of polyester and spandex while trying on and checking products at the stores. Data were collected by asking the staff at road specialty shops who has more professional knowledge about the advantages and functions of the existing products and recording their answers. Point color schemes in black color family in which body shapes can look beautiful were mostly checked. Due to the design characteristics that have to maintain the spine, the designs of all-in-one products that can maintain the upper and lower torsos were checked at underwear stores. The existing products were evaluated for their designs to be adopted including the material, integrated into the new design, and then modified and supplemented, and the following chiropractic apparel design was drawn (Table 1). An all-in-one design that wraps around the hips and upper thighs for easy counterbalancing between the upper and lower body was chosen. A design with an accent line that makes body shape looks beautiful and Used compression wear material that is comfortable for activities and presses tight against the skin. Activates the muscle activities of latissimus dorsi muscle using taping method, and provides tension to maintain correct posture of round back and shoulders. In conclusion, prevention, exercise, and stretching are important for scoliosis and sound posture for the loin should be constantly maintained. To that end, not only the proper use of chairs but also apparel that is worn all the time can be helpful

Unusual extraction treatment in Class II division 1 using C-orthodontic mini-implants.

AbstractThis paper describes the treatment of a female patient, aged 23 years and 5 months, with a Class II division 1 malocclusion, who showed severe anterior protrusion and lower anterior crowding. Specially-designed orthodontic mini-implants were placed bilaterally in the interdental space between both the upper and the lower posterior teeth. Both lower first molars showed severe apical lesions. Therefore, the treatment plan consisted of extraction of both upper first premolars and lower first molars, en masse retraction of the upper six anterior teeth, lower anterior alignment, and protraction of all the lower molars. C-implants® were used as substitutes for maxillary posterior anchorage teeth during anterior retraction and as hooks for mandibular molar protraction. The correct overbite and overjet were obtained by intruding and retracting the upper six anterior teeth into their proper positions. The dentition was detailed using conventional orthodontic appliances. The upper C-implants contributed to an improvement in facial balance, and the lower C-implants made it possible to protract the lower second and third molars with less effect on the axis of the lower anterior teeth. The active treatment period was 29 months and the patient's teeth continued to be stable 11 months after debonding

Fabrication of core/shell ZnWO4/carbon nanorods and their Li electroactivity

Carbon-coated ZnWO4 [C-ZW] nanorods with a one-dimensional core/shell structure were synthesised using hydrothermally prepared ZnWO4 and malic acid as precursors. The effects of the carbon coating on the ZnWO4 nanorods are investigated by thermogravimetry, high-resolution transmission electron microscopy, and Raman spectroscopy. The coating layer was found to be in uniform thickness of approximately 3 nm. Moreover, the D and G bands of carbon were clearly observed at around 1,350 and 1,600 cm-1, respectively, in the Raman spectra of the C-ZW nanorods. Furthermore, lithium electroactivities of the C-ZW nanorods were evaluated using cyclic voltammetry and galvanostatic cycling. In particular, the formed C-ZW nanorods exhibited excellent electrochemical performances, with rate capabilities better than those of bare ZnWO4 nanorods at different current rates, as well as a coulombic efficiency exceeding 98%. The specific capacity of the C-ZW nanorods maintained itself at approximately 170 mAh g-1, even at a high current rate of 3 C, which is much higher than pure ZnWO4 nanorods