Sequence optimization and multiple gene-targeting improve the inhibitory efficacy of exogenous double-stranded RNA against pepper mottle virus in Nicotiana benthamiana

Double-stranded RNA (dsRNA)-induced RNA interference is a promising agricultural technology for crop protection against various pathogens. Recent advances in this field have enhanced the overall efficiency with which this approach inhibits pathogenic viruses. Our previous study verified that treatment of Nicotiana benthamiana plants with dsRNAs targeting helper component-proteinase (HC-Pro) and nuclear inclusion b (NIb) genes protected the plant from pepper mottle virus (PepMoV) infection. The aim of this study was to improve the inhibitory efficacy of dsRNAs by optimizing the target sequences and their length and by targeting multiple genes via co-treatment of dsRNAs. Each of the two targeting dsRNAs were divided into three shorter compartments and we found that HC-Pro:mid-1st and NIb:mid-3rd showed significantly superior antiviral potency than the other fragments, including the parent dsRNA. In addition, we confirmed that the co-treatment of two dsRNAs targeting HC-Pro and NIb produced a greater inhibition of PepMoV replication than that obtained from individual dsRNA treatment. Complementing our previous study, this study will provide future directions for designing dsRNAs and enhancing their efficiency in dsRNA-mediated RNA interference technologies.This study was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MSIT) (No. 2021R1A5A1032428 and No. 2022R1A2C1011032). This was also supported by a grant from the New breeding technologies development Program (Project No. PJ01652102), Rural Development Administration, Republic of Korea

Lamellar keratoplasty using position-guided surgical needle and M-mode optical coherence tomography

Deep anterior lamellar keratoplasty (DALK) is an emerging surgical technique for the restoration of corneal clarity and vision acuity. The big-bubble technique in DALK surgery is the most essential procedure that includes the air injection through a thin syringe needle to separate the dysfunctional region of the cornea. Even though DALK is a well-known transplant method, it is still challenged to manipulate the needle inside the cornea under the surgical microscope, which varies its surgical yield. Here, we introduce the DALK protocol based on the position-guided needle and M-mode optical coherence tomography (OCT). Depth-resolved 26-gage needle was specially designed, fabricated by the stepwise transitional core fiber, and integrated with the swept source OCT system. Since our device is feasible to provide both the position information inside the cornea as well as air injection, it enables the accurate management of bubble formation during DALK. Our results show that real-time feedback of needle end position was intuitionally visualized and fast enough to adjust the location of the needle. Through our research, we realized that position-guided needle combined with M-mode OCT is a very efficient and promising surgical tool, which also to enhance the accuracy and stability of DALK

Differential patterns of nestin and glial fibrillary acidic protein expression in mouse hippocampus during postnatal development

Intermediate filaments, including nestin and glial fibrillary acidic protein (GFAP), are important for the brain to accommodate neural activities and changes during development. The present study examined the temporal changes of nestin and GFAP protein levels in the postnatal development of the mouse hippocampus. Mouse hippocampi were sampled on postnatal day (PND) 1, 3, 6, 18, and 48. Western blot analysis showed that nestin expression was high at PND 1 and markedly decreased until PND 18. Conversely, GFAP expression was acutely increased in the early phase of postnatal development. Nestin immunoreactivity was localized mainly in the processes of ramified cells at PND 1, but expression subsequently decreased. In contrast, GFAP was evident mainly in the marginal cells of the hippocampus at PND 1, but immunoreactivity revealed satellite, radial, or ramified shapes of the cells from PND 6-48. This study demonstrates that the opposing pattern of nestin and GFAP expressions in mouse hippocampus during postnatal development occur in the early development stage (PND 1-18), suggesting that the opposing change of nestin and GFAP in early postnatal development is important for neural differentiation and positioning in the mouse hippocampus

The expression and localization of inhibin isotypes in mouse testis during postnatal development

Inhibin, which is important for normal gonadal function, acts on the pituitary gonadotropins to suppress follicle-stimulating hormone (FSH) secretion. The level and cellular localization of the inhibin isotypes, α, βA and βB, in the testis of mice were examined during postnatal development in order to determine if inhibin expression is related to testicular maturation. Mouse testes were sampled on postnatal days (PNDs) 1, 3, 6, 18, 48 and 120, and analyzed by Western blotting and immunofluorescence. Western blot analysis showed very low levels of inhibin α, βA and βB expression in the testes at days 1 to 6 after birth. The levels then increased gradually from PND 18 to 48-120, and there were significant peaks at PND 48. Inhibin α, βA and βB were detected in testicular cells during postnatal development using immunohistochemistry. The immunoreactivity of inhibin α was rarely observed in testicular cells during PND 1 to 6, or in the cytoplasmic process of Sertoli cells surrounding the germ cells and interstitial cells during PND 18 to 120. Inhibin βA and βB immunoreactivity was rarely observed in the testis from PND 1 to 6. On the other hand, it was observed in some spermatogonial cells, as well as in the interstitial space between PND 48 and PND 120. We conclude that the expression of inhibin isotypes increases progressively in the testis of mice with increasing postnatal age, suggesting that inhibin is associated with a negative feedback signal for FSH in testicular maturation

Endotracheal intubation in rabbits using a video laryngoscope with a modified blade

Rabbits are being increasingly used as companion animals, and in research; thus, the need for proper veterinary care for rabbits has increased. Surgical access is more challenging in rabbits under inhalation anesthesia compared to other animals, such as dogs and cats. Rabbits have a very narrow and deep oral cavity, large incisors, and a large tongue. Moreover, their temporomandibular joint has limited mobility, making it more difficult to approach the larynx. Various methods have been proposed to overcome this difficulty. The video laryngoscope was introduced in 1999 and is useful when airway intubation is unsuccessful using a conventional laryngoscope. We postulated that a video laryngoscope with a modified size 1 Macintosh blade (McGrath MAC Video Laryngoscope, Medtronic, USA) would facilitate the intubation of New Zealand White rabbits. Sixteen specific-pathogen-free male New Zealand White rabbits weighing 3.45–4.70 kg were studied. All rabbits were intubated using the video laryngoscope. Typically, a 3.0 mm endotracheal tube was used for rabbits weighing  4 kg. During surgery, anesthesia was well maintained, and there were no major abnormalities in the animals’ conditions. No rabbit developed breathing difficulties or anorexia after recovering from anesthesia. We established an intubation method using a video laryngoscope with a modified blade and stylet in the supine (ventrodorsal) position and successfully applied it in 16 rabbits. It is useful for training novices and for treating rabbits in veterinary hospitals with few staff members and animal research facilities where there are insufficient human resources

NICE 2023 Zero-shot Image Captioning Challenge

In this report, we introduce NICE project\footnote{\url{https://nice.lgresearch.ai/}} and share the results and outcomes of NICE challenge 2023. This project is designed to challenge the computer vision community to develop robust image captioning models that advance the state-of-the-art both in terms of accuracy and fairness. Through the challenge, the image captioning models were tested using a new evaluation dataset that includes a large variety of visual concepts from many domains. There was no specific training data provided for the challenge, and therefore the challenge entries were required to adapt to new types of image descriptions that had not been seen during training. This report includes information on the newly proposed NICE dataset, evaluation methods, challenge results, and technical details of top-ranking entries. We expect that the outcomes of the challenge will contribute to the improvement of AI models on various vision-language tasks.Comment: Tech report, project page https://nice.lgresearch.ai

Muscular loads associated with floor vacuuming with cordless stick cleaners

The muscular loads of the upper extremity and chest muscles were assessed during floor vacuuming with stick vacuum cleaners. Ten participants conducted vacuuming with two stick cleaners (weight: 3.03 kg, </jats:p><jats:p> 3.56 kg) on tiled and carpeted floors while the myoelectric activity of the upper extremity and chest muscles were collected. The mean muscle activation levels ranged from 15.5% to 45.7% of the maximum voluntary capacity. Greater muscular loads were observed on the forearm and shoulder muscles when vacuuming with repeated curved strokes (vs. straight storkes), and on the chest muscle when vacuuming the carpeted floor. The range of muscular loads were higher than that of occupational tasks such as house painting and dental drilling, which have been known as intensive activities for upper extremities. The results suggest that floor vacuuming with stick vacuum cleaners is physically demanding for household consumers. Ergonomic interventions such as more user-friendly design should be considered

Foot kinematics and leg muscle activation patterns are altered in those with limited ankle dorsiflexion range of motion during incline walking

Background Larger ankle dorsiflexion (DF) is required when walking on inclined surfaces. Individuals with limited DF range of motion (ROM) may experience greater tissue stress on sloped surfaces and walk in altered gait patterns compared to the those with normal DF ROM. Research question Would the individuals with limited DF ROM walk with distinctive ankle DF patterns compared to those with normal DF ROM on the inclined surfaces? Methods Ten Limited DF ROM (passive ROM=35.3 ?? 2.7??) and nine Normal DF ROM (passive ROM=46.4 ?? 4.2??) participants walked on a treadmill at five slope angles (0??, 5??, 10??, 15??, 20??) for 2 min at a self-selected speed. The peak DF angles and the peak myoelectric activity levels of the tibialis anterior (TA) and soleus (SOL) muscles were quantified during the swing and stance phases of each walking trial, and they were compared between the two groups. Results Participants with limited DF ROM walked with smaller peak DF (3.1?? at 0?? slope ~ 8.4?? at 20?? slope) and greater peak TA activity in swing than those of the Normal ROM participants (3.4?? ~ 12.2??), with significant differences at 20?? slope. The peak DF angle in stance (Limited: 9.6?? ~ 19.0??; Normal: 10.1?? ~ 21.0??) did not differ between the two groups at all slopes, but the peak activity of the SOL muscle was significantly greater for the Limited group at slopes of 10?? and higher. Significance Study results indicate that incline walking could be more challenging to the individuals with limited DF ROM as they need to approach and push-off the sloped surfaces with more efforts of the dorsiflexor and the plantar flexor muscles, respectively. Prolonged walking on inclined surfaces may produce faster development of muscle fatigue or tissue damage than those with normal DF ROM