A 3D Printable Electroconductive Biocomposite Bioink Based on Silk Fibroin-Conjugated Graphene Oxide.

Reduced graphene oxide (rGO) has wide application as a nanofiller in the fabrication of electroconductive biocomposites due to its exceptional properties. However, the hydrophobicity and chemical stability of rGO limit its ability to be incorporated into precursor polymers for physical mixing during biocomposite fabrication. Moreover, until now, no suitable rGO-combining biomaterials that are stable, soluble, biocompatible, and 3D printable have been developed. In this study, we fabricated digital light processing (DLP) printable bioink (SGOB1), through covalent reduction of graphene oxide (GO) by glycidyl methacrylated silk fibroin (SB). Compositional analyses showed that SGOB1 contains approximately 8.42% GO in its reduced state. Our results also showed that the rGO content of SGOB1 became more thermally stable and highly soluble. SGOB1 hydrogels demonstrated superior mechanical, electroconductive, and neurogenic properties than (SB). Furthermore, the photocurable bioink supported Neuro2a cell proliferation and viability. Therefore, SGOB1 could be a suitable biocomposite for neural tissue engineering.

Correction to: Silk Fibroin Bioinks for Digital Light Processing (DLP) 3D Bioprinting.

Correction to: Chapter 4 in: H. J. Chun et al. (eds.), Bioinspired Biomaterials, Advances in Experimental Medicine and Biology 1249, https://doi.org/10.1007/978-981-15-3258-0_4.

Silk Fibroin Bioinks for Digital Light Processing (DLP) 3D Bioprinting.

Three-dimensional (3D) bioprinting has been a highly influential technology in the field of tissue engineering to enable speedy and precise spatial patterning of cells, growth factors, and biomaterials. Bioink is one of the main factors in 3D bioprinting, and hydrogels are excellent matrix type by means of bioinks for 3D bioprinting. Recently, stereolithographic bioprinting via digital light processing (DLP) that allows high spatial resolution and rapid printing time of complex structures has attracted many studies. However, a small number of bioinks have been applied to DLP bioprinting in comparison with bioinks for other bioprinters. We developed a novel bioink based on silk fibroin that has been extensively used in biomedical fields due to its positive biological and biochemical properties as biomaterials. In this chapter, we summarized the silk fibroin basics and various applications of silk fibroin as printing material. Also, fabrication and performance of silk-based bioink for DLP bioprinter were discussed.

BPH1, a novel substrate receptor of CRL3, plays a repressive role in ABA signal transduction.

KEY MESSAGE: BPH1 acts as a substrate receptor of CRL3 complex and negatively regulates ABA-mediated cellular responses. The study on its function provides information that helps further understand the relationship between ABA signaling and UPS. Abscisic acid (ABA) plays a crucial role in a variety of cellular processes, including seed dormancy, inhibition of seedling growth, and drought resistance in plants. Cullin3-RING E3 ligase (CRL3) complex is a type of multi-subunit E3 ligase, and BTB/POZ protein, a component of CRL3 complex, functions as a receptor to determine a specific substrate. To elucidate the CRL3 complex that participates in ABA-mediated cellular processes, we first investigated ABA-inducible BTB/POZ genes based on data from the AtGenExpress Visualization Tool (AVT). We then isolated an ABA-inducible gene encoding a potential CRL3 substrate receptor in Arabidopsis, BPH1 (BTB/POZ protein hypersensitive to ABA 1). The isolate gene has a BTB/POZ domain and a NPH3 domain within its N-terminal and C-terminal region, respectively. Yeast two-hybrid and co-immunoprecipitation assays showed that BPH1 physically interacted with cullin3a, a scaffold protein of CRL3, suggesting that it functions as an Arabidopsis CRL3 substrate receptor. The functional mutation of BPH1 caused delayed seed germination in response to ABA and enhanced sensitivity by NaCl and mannitol treatments as ABA-related stresses. Moreover, bph1 mutants exhibited enhanced stomatal closure under ABA application and reduced water loss when compared with wild-type, implying their enhanced tolerance to drought stress. Based on the information from microarray/AVT data and expression analysis of various ABA-inducible genes between wild-type and bph1 plants following ABA treatments, we concluded loss of BPH1 resulted in hyper-induction of a large portion of ABA-inducible genes in response to ABA. Taken together, these results show that BPH1 is negatively involved in ABA-mediated cellular events.

Silk Fibroin in Wound Healing Process.

Silk fibroin (SF), a natural bioproduct, has been extensively used in biological and biomedical fields including wound healing due to its robust biocompatibility, less immunogenic, non-toxic, non-carcinogenic, and biodegradable properties. SF in different morphologic forms, such as hydrogels, sponges, films, electrospun nanofiber mats, and hydrocolloid dressings, have been successfully used for therapeutic use as wound dressings to induce the healing process. SF has also been known to promote wound healing by increasing the cell growth, proliferation, and migration of different cells types involved in the different phase of wound healing process. In this review, we summarize the different morphologic forms of SF that have been used in the treatment of various wound healing process. We also discuss the effect of SF on various cells types during the SF-induced healing process. Furthermore, we highlight molecular signaling aspects of the SF-induced healing process.

Effects of a work-based critical reflection program for novice nurses.

BACKGROUND: Critical reflection is effective in improving students' communication abilities and confidence. The aim of this study was to evaluate the effectiveness of a work-based critical reflection program to enhance novice nurses' clinical critical-thinking abilities, communication competency, and job performance. METHODS: The present study used a quasi-experimental design. From October 2014 to August 2015, we collected data from 44 novice nurses working in an advanced general hospital in S city in Korea. Nurses in the experimental group participated in a critical reflection program for six months. Outcome variables were clinical critical-thinking skills, communication abilities, and job performance. A non-parametric Mann-Whitney U-test and a Wilcoxon rank sum test were selected to evaluate differences in mean ranks and to assess the null hypothesis that the medians were equal across the groups. RESULTS: The results showed that the clinical critical-thinking skills of those in the experimental group improved significantly (p = 0.003). The differences in mean ranks of communication ability between two groups was significantly statistically different (p = 0.028). Job performance improved significantly in both the experimental group and the control group, so there was no statistical difference (p = 0.294). CONCLUSIONS: We therefore suggest that a critical reflection program be considered an essential tool for improving critical thinking and communication abilities among novice nurses who need to adapt to the clinical environment as quickly as possible. Further, we suggest conducting research into critical reflection programs among larger and more diverse samples.

The effect of chest expansion exercise with TENS on gait ability and trunk control in chronic stroke patients.

[Purpose] The purpose of this study was to investigate the effect of chest expansion exercise with transcutaneous electrical nerve stimulation (TENS) on gait ability and trunk control of patient with stroke. [Subjects and Methods] The subjects were divided into 7 in the chest expansion exercise with TENS group (experimental group) and 7 in the chest expansion exercise with placebo TENS (control group). The gait ability and trunk contol were measured using Six-Minute Walk Test, Tinetti gait index and Trunk impairment scale (TIS). [Results] Both the experimental group and the control group showed significant improvement in the Six-Minute Walk Test, Tinetti gait index, and TIS total score. The dynamic sitting balance and coordination of TIS showed significant improvement only in the experimental group. In comparison between the two groups, the experimental group showed a more significant improvement in Tinetti gait index and TIS total score than the control group. [Conclusion] This study showed that chest expansion exercise with TENS was an effective method for improving gait ability and trunk control in chronic stroke patients.

Artificial Auricular Cartilage Using Silk Fibroin and Polyvinyl Alcohol Hydrogel.

Several methods for auricular cartilage engineering use tissue engineering techniques. However, an ideal method for engineering auricular cartilage has not been reported. To address this issue, we developed a strategy to engineer auricular cartilage using silk fibroin (SF) and polyvinyl alcohol (PVA) hydrogel. We constructed different hydrogels with various ratios of SF and PVA by using salt leaching, silicone mold casting, and freeze-thawing methods. We characterized each of the hydrogels in terms of the swelling ratio, tensile strength, pore size, thermal properties, morphologies, and chemical properties. Based on the cell viability results, we found a blended hydrogel composed of 50% PVA and 50% SF (P50/S50) to be the best hydrogel among the fabricated hydrogels. An intact 3D ear-shaped auricular cartilage formed six weeks after the subcutaneous implantation of a chondrocyte-seeded 3D ear-shaped P50/S50 hydrogel in rats. We observed mature cartilage with a typical lacunar structure both in vitro and in vivo via histological analysis. This study may have potential applications in auricular tissue engineering with a human ear-shaped hydrogel.

The immediate effects of rib cage joint mobilization and chest wall stretch on muscle tone and stiffness of respiratory muscles and chest expansion ability in patients with chronic stroke.

[Purpose] The main purpose of this study was to identify the impact of rib cage joint mobilization and chest wall stretch on respiratory muscle tone and stiffness and chest expansion in stroke patients and to compare the effects of both interventions. [Subjects and Methods] Subjects were randomly assigned to a rib cage joint mobilization group (n=15) or a chest wall stretch group (n=15). Respiratory muscle tone and stiffness were measured using a myotonometer, and the chest expansion was gauged using a measuring tape. [Results] A significant difference was found on comparing the respiratory muscle tone and stiffness on the affected and sound side before intervention. Although both groups showed an increase in respiratory muscle tone and stiffness after intervention, no significant difference was found. A significant increase in chest expansion was observed; however, no significant difference was observed in the variations between the groups. [Conclusion] This study suggests that rib cage joint mobilization and chest wall stretch exercises can be used to increase chest expansion potential and respiratory muscle tone in patients with chronic stroke.

ABA-HYPERSENSITIVE BTB/POZ PROTEIN 1 functions as a negative regulator in ABA-mediated inhibition of germination in Arabidopsis.

To elucidate the contribution of CRL3-ABA-mediated responses, we attempted to find CRL3 substrate receptors involved in ABA signaling. One gene named ABA-HYPERSENSITIVE BTB/POZ PROTEIN 1 (AHT1) was upregulated more than 2.5 times by ABA, and its coding region possessed a BTB/POZ domain, which is the common feature of CRL3 substrate receptors. Loss of AHT1 led to retardation of the germination process, not inhibition of root growth. AHT1 transcripts also increased in response to mannitol, NaCl and drought treatments at the seedling stage and in dry seeds. High expression of AHT1 in dry seeds was inhibited by the defect of ABA signaling components such as ABI1, ABI3 and SRKs indicating that the expression of AHT1 is dependent on ABA signaling. Among bZIP transcription factors participating in ABA signaling, the losses of ABI5/DPBF1, AREB1/ABF2, EEL/DPBF4 and DPBF2/bZIP67 resulted in reduced AHT1 expression, showing that these transcription factors play a positive role in ABA-induced AHT1 expression. While loss of AHT1 did not affect the expression pattern of NCED3, ABI2, SRKs and AREB/ABF genes, it led to hyperinduction of ABI5/DPBF genes such as ABI5/DPBF1, EEL/DPBF4 and AREB3/DPBF3, which are mainly involved in seed development and germination, as well as ABA-inducible genes transactivated by ABI5. Overall, these findings indicate that AHT1 negatively regulates ABA-mediated inhibition of germination, possibly by repressing the expression of a subset of ABI5/DPBF subfamily genes, and that AHT1 may be regulated by a negative feedback process through its linkage with a part of ABI5/DPBF proteins.

Autophagy-related proteins are required for degradation of peroxisomes in Arabidopsis hypocotyls during seedling growth.

Plant peroxisomes play a pivotal role during postgerminative growth by breaking down fatty acids to provide fixed carbons for seedlings before the onset of photosynthesis. The enzyme composition of peroxisomes changes during the transition of the seedling from a heterotrophic to an autotrophic state; however, the mechanisms for the degradation of obsolete peroxisomal proteins remain elusive. One candidate mechanism is autophagy, a bulk degradation pathway targeting cytoplasmic constituents to the lytic vacuole. We present evidence supporting the autophagy of peroxisomes in Arabidopsis thaliana hypocotyls during seedling growth. Mutants defective in autophagy appeared to accumulate excess peroxisomes in hypocotyl cells. When degradation in the vacuole was pharmacologically compromised, both autophagic bodies and peroxisomal markers were detected in the wild-type vacuole but not in that of the autophagy-incompetent mutants. On the basis of the genetic and cell biological data we obtained, we propose that autophagy is important for the maintenance of peroxisome number and cell remodeling in Arabidopsis hypocotyls.

Effects of low-level laser therapy, electroacupuncture, and radiofrequency on the pigmentation and skin tone of adult women.

[Purpose] In this study, the effects of low-level laser therapy (LLLT), electroacupuncture (EA), and radiofrequency (RF), which are used in physical therapy, on the pigmentation and skin tone of adult women's faces were investigated to provide basic data for skin interventions. [Subjects and Methods] Thirty adult females were assigned to either an LLLT group (n=10), an EA group (n=10), or an RF group (n=10). The intervention was performed in two 15-minute sessions per week for six weeks. Subjects' skin tone and pigmentation were observed before and after the intervention. [Results] The EA group showed significant reductions in pigmentation in the left and right eye rims, as well as in the left cheek. The RF group showed significant post-intervention reductions in pigmentation under the left eye, as well as in the left and right eye rims and the left cheek. The LLLT group showed significant increases in skin tone in the forehead and both eye rims. The RF group showed significant increases in skin tone under both eyes. [Conclusion] The application of LLLT, EA, and RF had positive effects on pigmentation and skin tone of adult women's faces.

The effects of sole vibration stimulation on Korean male professional volleyball players' jumping and balance ability.

[Purpose] The purpose of this study was to investigate differences in jumping ability and lower limb balance ability elicited by plyometric training and vibration exercise, of volleyball players with and without ankle injuries, which frequently occur among Korean professional volleyball players. [Subjects and Methods] Twenty-eight volleyball players were divided into the following groups: plyometric with ankle injury (PAI) group; plyometric with non-ankle injury (PAN) group; vibrator with ankle injury (VAI) group; and vibrator with non-ankle injury (VAN) group. After exercise and whole body vibration stimulation, their vertical jumping abilities, side step, and static equilibrium ability were measured. [Results] The vibration exercise group which had experienced ankle injuries showed significant improvements in the sidestep test after the intervention compared to before the intervention. In vertical jumping as well, significant improvements were observed in the VAI group and the VAN group following vibration exercise. In the balance ability test, significant improvesments in the PAN group and the PAI group were observed after the intervention. According to the results of the right side, there was significant change in the left/back side test and the right/back side test before and after the intervention; and in the test of one-leg standing with eyes closed, there were significant group, timing, and interaction effects. [Conclusions] The training method which effectively improved the jumping ability of volleyball players was plyometric training, and for balance ability improvement, whole body vibration exercise was effective.

Effect of eye movements and proprioceptive neuromuscular facilitation on balance and head alignment in stroke patients with neglect syndrome.

[Purpose] The purpose of this study was to assess the effect of eye movements and proprioceptive neuromuscular facilitation (PNF) on patients with neglect syndrome. [Subjects and Methods] The subjects were randomly allocated to 2 groups: the eye movements (EM) group; and the PNF with eye movements (PEM) group. The program was conducted five times each week for 6 weeks. Balance (both static and dynamic) and head alignment (craniovertebral angle and cranial rotation angle) were measured before and after testing. [Results] In measurements of static balance, the EM group showed significant improvement in sway length and sway area when examined in the eyes-open condition, but not when examined in the eyes-closed condition. The PEM group showed significant improvement when examined under both conditions. In the assessment of dynamic balance, both groups showed significant improvement in measurements of sway areas. With respect to head alignment, there were no significant differences pre- and post-testing in either the craniovertebral angle or the cranial rotation angle in the EM group, but the PEM group showed significant differences in both measurements. [Conclusion] These results suggest that in stroke patients with neglect syndrome, PNF with eye movements, rather than eye movements alone, has a greater positive effect on balance and head alignment.

Two-point discrimination of the upper extremities of healthy Koreans in their 20's.

[Purpose] The present study attempted to measure two-point discrimination in the upper extremities of healthy Koreans in their 20's. [Subjects and Methods] Using a three-point esthesiometer, we conducted an experiment with a group of 256 college students (128 male and 128 female), attending N University in Chonan, Republic of Korea. [Results] Females showed two-point discrimination at a shorter distance than males at the following points: (i) 5 cm above the elbow joint, the middle part, and 5 cm below the shoulder joint of the anterior upper arm; (ii) 5 cm above the elbow joint and 5 cm below the shoulder joint of the posterior upper arm; (iii) 5 cm above the front of the wrist joint of the forearm; 5 cm below the elbow joint, the palmar part of the distal interphalangeal joint of the thumb, the dorsal part of the distal interphalangeal joint of the middle and little fingers. It was also found that females showed greater two-point discrimination than males in distal regions rather than proximal regions. [Conclusion] The findings of this study will help establish normal values for two-point discrimination of upper extremities of young Koreans in their 20's.

DWD HYPERSENSITIVE TO UV-B 1 is negatively involved in UV-B mediated cellular responses in Arabidopsis.

Among T-DNA insertion mutants of various cullin4-RING ubiquitin E3 ligase (CRL4) substrate receptors, one mutant that exhibits enhanced sensitivity in response to ultraviolet-B (UV-B) illumination has been isolated and its corresponding gene has been named DWD HYPERSENSITIVE TO UV-B 1 (DHU1) in Arabidopsis. dhu1 lines showed much shorter hypocotyls than those in wild type under low doses of UV-B. Other light did not alter hypocotyl growth patterns in dhu1, indicating the hypersensitivity of dhu1 is restricted to UV-B. DHU1 was upregulated by more than two times in response to UV-B application of 1.5 µmol m(-2) s(-1), implying its possible involvement in UV-B signaling. DHU1 is able to bind to DDB1, an adaptor of CRL4; accordingly, DHU1 is thought to act as a substrate receptor of CRL4. Microarray data generated from wild-type and dhu1 under low doses of UV-B revealed that 209 or 124 genes were upregulated or downregulated by more than two times in dhu1 relative to wild type, respectively. About 23.4 % of the total upregulated genes in dhu1 were upregulated by more than five times in response to UV-B based on the AtGenExpress Visualization Tool data, while only about 1.4 % were downregulated to the same degree by UV-B, indicating that loss of DHU1 led to the overall enhancement of the upregulation of UV-B inducible genes. dhu1 also showed altered responsiveness under high doses of UV-B. Taken together, these findings indicate that DHU1 is a potent CRL4 substrate receptor that may function as a negative regulator of UV-B response in Arabidopsis.

Mitochondrial control region variation in a Korean population sample.

Mitochondrial DNA (mtDNA) control region (16024-576) sequences were generated from 281 individuals from South Korea. Robotic liquid handling, a redundant sequencing strategy, and a series of quality control checks were implemented to ensure the high quality of the dataset. This population sample showed a low random match probability (0.25 %) and high genetic diversity (0.9933). The haplogroup breakdown was consistent with previous studies describing Korean mtDNA variation. The 224 unique haplotypes (33 shared) presented will supplement the data already publically available.

Thoracic coupled motions of korean men in good health in their 20s.

[Purpose] The purpose of this study was to investigate thoracic coupled motions of 20 Korean young individuals. [Methods] Thoracic motion of twenty healthy male college students aged 23.2±3.1 was examined. The coupled motions of the thoracic regions T1-4, T4-8, T8-12 were measured using a three dimensional motion capture system. [Results] Coupled axial rotation in the same direction as lateral bending was observed in T1-T4 and T4-T8 in the neutral, flexed, and extended postures of the thoracic spine. In T8-T12, coupled axial rotation in the same direction as lateral bending were observed in the neutral and flexed postures, while coupled axial rotation in the opposite direction was observed in an extended posture. [Conclusion] The patterns of coupled motions in the thoracic spine demonstrated some variability between postures and regions in vivo. However, coupled motions in the same direction were predominantly lateral flexion or axial rotation in the three postures.

Capsaicin stimulates glucose uptake in C2C12 muscle cells via the reactive oxygen species (ROS)/AMPK/p38 MAPK pathway.

Capsaicin has been reported to regulate blood glucose levels and to ameliorate insulin resistance in obese mice. This study demonstrates that capsaicin increases glucose uptake directly by activating AMP-activated protein kinase (AMPK) in C2C12 muscle cells, which manifested as an attenuation of glucose uptake when compound C, an AMPK inhibitor, was co-administered with capsaicin. However, the insulin signaling molecules insulin receptor substrate-1 (IRS-1) and Akt were not affected by capsaicin. Additional results showed that p38 mitogen-activated protein kinase (MAPK) is also involved in capsaicin-induced glucose transport downstream of AMPK because capsaicin increased p38 MAPK phosphorylation significantly and its specific inhibitor SB203580 inhibited capsaicin-mediated glucose uptake. Treatment with an AMPK inhibitor reduced p38 MAPK phosphorylation, but the p38 MAPK inhibitor had no effect on AMPK. Capsaicin stimulated ROS generation in C2C12 muscle cells, and when ROS were captured using the nonspecific antioxidant NAC, the increase in both capsaicin-induced AMPK phosphorylation and capsaicin-induced glucose uptake was attenuated, suggesting that ROS function as an upstream activator of AMPK. Taken together, these results suggest that capsaicin, independent of insulin, increases glucose uptake via ROS generation and consequent AMPK and p38 MAPK activations.

Functional Skeletal Muscle Regeneration Using Muscle Mimetic Tissue Fabricated by Microvalve-Assisted Coaxial 3D Bioprinting.

3D-printed artificial skeletal muscle, which mimics the structural and functional characteristics of native skeletal muscle, is a promising treatment method for muscle reconstruction. Although various fabrication techniques for skeletal muscle using 3D bio-printers are studied, it is still challenging to build a functional muscle structure. A strategy using microvalve-assisted coaxial 3D bioprinting in consideration of functional skeletal muscle fabrication is reported. The unit (artificial muscle fascicle: AMF) of muscle mimetic tissue is composed of a core filled with medium-based C2C12 myoblast aggregates as a role of muscle fibers and a photo cross-linkable hydrogel-based shell as a role of connective tissue in muscles that enhances printability and cell adhesion and proliferation. Especially, a microvalve system is applied for the core part with even cell distribution and strong cell-cell interaction. This system enhances myotube formation and consequently shows spontaneous contraction. A multi-printed AMF (artificial muscle tissue: AMT) as a piece of muscle is implanted into the anterior tibia (TA) muscle defect site of immunocompromised rats. As a result, the TA-implanted AMT responds to electrical stimulation and represents histologically regenerated muscle tissue. This microvalve-assisted coaxial 3D bioprinting shows a significant step forward to mimicking native skeletal muscle tissue.