Complete mitochondrial genome of the southern painted turtle (Chrysemys dorsalis, Testudines: Emydidae) in Korea.

The complete mitochondrial genome of Chrysemys dorsalis in Korea was sequenced and characterized. The mitochondrial genome is 17,258 bp in length and the GC content is 39%. It is constituted of 37 genes, 13 protein-coding genes, 22 transfer RNA genes, 2 ribosomal RNA genes, and a noncoding region. Phylogenetic analysis reveals that C. dorsalis forms a monophyletic group with C. picta turtles but is distinctly separated from them, aligning with previous findings. In Korea, C. dorsalis forms a discrete clade, separate from both native and invasive turtle species. No evidence of genetic disturbance or intermingling is observed. This is the first case of a complete mitochondrial genome from C. dorsalis and provides crucial data for understanding C. dorsalis and managing invasive species effectively, emphasizing the need for continued mitochondrial genome data accumulation.

The Donut DIEP Flap: A Novel Scarless Technique.

Deep inferior epigastric perforator (DIEP) flap surgery is a common technique for breast reconstruction. However, the long and noticeable abdominal scarring is one of its greatest disadvantages. Here, we suggest a minimal scarring DIEP flap harvest with a novel abdominal design. The key to this method is to preserve more skin tissue with a circular design centered around the navel. Three circular incisions are made: (1) the smallest circle represents an incision around the umbilicus; (2) the middle circle with a radius of <3.5 cm indicates skin incision because the donut-like skin paddle between the small and middle circles is included in the flap; (3) the largest circle with a radius of >10 cm represents the extent of flap dissection that was done diagonally from the skin toward the Scarpa's fascia. Pedicle dissection is proceeded with the conventional method. For closure, a purse-string suture is performed with the umbilicus at its center leaving only a single small circular scar around the umbilicus as the final scar. Closure of the donor-site is possible with the patient in a supine or slightly flexed position due to less skin excision. LEVEL OF EVIDENCE: Therapeutic, V.

Age Group Classification of Dental Radiography without Precise Age Information Using Convolutional Neural Networks.

Automatic age estimation using panoramic dental radiographic images is an important procedure for forensics and personal oral healthcare. The accuracies of the age estimation have increased recently with the advances in deep neural networks (DNN), but DNN requires large sizes of the labeled dataset which is not always available. This study examined whether a deep neural network is able to estimate tooth ages when precise age information is not given. A deep neural network model was developed and applied to age estimation using an image augmentation technique. A total of 10,023 original images were classified according to age groups (in decades, from the 10s to the 70s). The proposed model was validated using a 10-fold cross-validation technique for precise evaluation, and the accuracies of the predicted tooth ages were calculated by varying the tolerance. The accuracies were 53.846% with a tolerance of ±5 years, 95.121% with ±15 years, and 99.581% with ±25 years, which means the probability for the estimation error to be larger than one age group is 0.419%. The results indicate that artificial intelligence has potential not only in the forensic aspect but also in the clinical aspect of oral care.

Prophylactic intraoperative wound irrigation with antibiotic solution for the prevention of surgical incisional wound infections: Systematic literature review and meta-analysis.

BACKGROUND: There is no consensus on the use of prophylactic intraoperative wound irrigation (pIOWI) for reducing surgical-site infections (SSIs), particularly surgical incisional wounds, and opinions are divided on whether an antibiotic should be mixed with the solution used for incisional wound irrigation. No large, reliable cohort studies or meta-analyses of pIOWI for surgical incisional wounds were found in PubMed or Embase. METHODS: We searched the PubMed, Embase, Cochrane Library, Insight, Scopus, and Web of Science databases for primary research articles investigating the relevance of pIOWI with antibiotics for SSIs. The initial search was limited to human studies published before January 2022, which were indexed as randomized controlled trials (RCTs); clinical trials; or observational, cross-sectional, or cohort studies. We used R statistical software version 4.1.2 for this meta-analysis. RESULTS: The odds ratio of the random-effects model was 0.519 [95% confidence interval (CI); 0.311, 0.864, P-value < 0.0117], indicating that the intervention had a statistically significant effect on surgical complications. To examine the heterogeneity of the entire study, we performed statistical analysis (τ2 (tau square) = 0.4175 [0.0732; 2.8178]; τ (tau) = 0.6461 [0.2706; 1.6786]; Higgins' I2 = 64.0% [34.8%; 80.1%]; and H = 1.67 [1.24; 2.24]). CONCLUSIONS: Many studies have shown that inclusion of antibiotics in the irrigation solution significantly lowers the rate of soft-tissue-related SSIs, but those studies contain both heterogeneity and many biases. Further large, prospective RCTs only limited to incisional SSIs and excluding other variables and biases in the field of plastic surgery are needed.

Late reconstruction of post-traumatic enophthalmos and hypoglobus using three-dimensional implants: a case series.

Post-traumatic enophthalmos and hypoglobus are common sequelae of facial bone fractures, even after reduction surgery. They are associated with functional and esthetic issues, which may lower the quality of life. These deformities frequently present late, and adequate correction is difficult. We report three cases of late inferior orbital rim reconstructions with three-dimensional printed implants to help resolve these problems. The average duration between the traumatic event and surgery was 3 years and 4 months. One patient was treated with a completely absorbable implant and exhibited satisfactory results until the implant started to biodegrade at 1 year and 9 months after surgery. Two patients were treated with a permanent implant and demonstrated satisfactory results. However, longer follow-up periods were needed. There were no complications such as infection, diplopia, or restriction of ocular motion and the patients were satisfied with the esthetic results.

Cranioplasty and temporal hollowing correction with a three-dimensional printed bioresorbable mesh and double vertical suture anchor muscle sling: a case report.

Postoperative temporal hollowing is a common complication of craniotomy. Damage and repositioning of the temporalis muscle can lead to a depression in the temporal side of the skull with inferior bulging, worsening aesthetic outcomes. We report a case of cranioplasty with three-dimensional (3D) printed mesh involving an additional correction using a temporalis muscle sling to help address this problem. A 3D-printed bioabsorbable mesh was prepared based on preoperative facial computed tomography, and was fixed to the hollowed area for tissue augmentation. The temporalis muscle was elevated and fanned out to its original position, and a sling was attached to a screw that was fixed to the mesh. For reinforcement, an additional sling was attached to another screw fixed to the mesh 2-3 cm vertically above the first screw. Aesthetic results were confirmed immediately after surgery and later during outpatient follow-up. Both depression and lateral bulging were resolved, and there was no delayed drooping of the temporalis muscle on 6-month follow-up. There were no complications, and the patient was satisfied with the appearance. This is a simple yet effective technique with a low risk of complications, and should be considered for postoperative temporal hollowing patients, especially those with severe lateral bulging.

Risk assessment for the native anurans from an alien invasive species, American bullfrogs (Lithobates catesbeianus), in South Korea.

The invasive species are of global concern, and the Invasive American Bullfrog (IAB; Lithobates catesbeianus) is one of the worst invasive amphibian species worldwide. Like other countries, South Korea is also facing challenges from IAB. Although many studies indicated impacts of IAB on native anurans in Korea, the actual risk at the specific level is yet to evaluate. Considering the putative invasiveness of IAB, it is hypothesized that any species with the possibility of physical contact or habitat sharing with them, will have a potential risk. Thus, we estimated and observed their home range, preferred habitats, morphology, behavior, and ecology. Then, comparing with existing knowledge, we assessed risks to the native anurans. We found a home range of 3474.2 ± 5872.5 m2 and identified three types of habitats for IAB. The analyses showed at least 84% of native anurans (frogs and toads) were at moderate to extreme risks, which included all frogs but only 33% of toads. Finally, we recommended immediate actions to conserve the native anurans based on our results. As this study is the first initiative to assess the specific risk level from the invasiveness of L. catesbeianus, it will help the managers to set conservation priorities and strategies.

Hydrogel Stamping for Rapid, Multiplexed, Point-of-Care Immunostaining of Cells and Tissues.

In the era of precision oncology, multicolor fluorescence imaging has become a core technology for multiplexed molecular analysis of cellular and tissue specimens. However, conventional solution-based staining is labor-intensive and time-consuming and requires considerable expertise to yield optimal results, which creates difficulties for employing this technology in resource-limited settings. Here, we report a new immunostaining method based on hydrogel stamping, which is simple, fast, easy to use, and reproducible. We showed that a hydrophilic hydrogel stamp could effectively transfer fluorescent antibodies to targets and withdraw an excess solution when the reaction is completed, obviating the need for extra washing. This unique property allows for quality immunostaining in 5 min for cells using one-eighth of antibody consumption compared to the conventional solution-based method. Furthermore, we implemented fluorescence quenching and immunocycling with hydrogel staining for multiplexed analysis of 9 protein markers at a single cell level. Finally, we applied the immunocycling method to human breast cancer tissue samples and showed quality immunostaining over a large area (∼2 cm2) in 30 min for molecular subtyping of breast cancer. The hydrogel immunostaining could open new opportunities for rapid, automated, and multiplexed profiling in compact point-of-care systems for molecular cancer diagnosis.

Robotic DIEP Flap Harvest through a Totally Extraperitoneal Approach Using a Single-Port Surgical Robotic System.

SUMMARY: Deep inferior epigastric artery perforator (DIEP) flap surgery is one of the most common techniques for breast reconstruction using abdominal tissue. Although the DIEP flap reduces donor-site morbidity when compared with the conventional free transverse rectus abdominis musculocutaneous flap, violation of the anterior rectus sheath, rectus muscle, and motor nerves cannot be avoided. To further minimize tissue damage in the donor site, the use of a surgical robotic system for DIEP flap harvest with a totally extraperitoneal approach has been suggested. This totally extraperitoneal approach has a long learning curve because of the narrow preperitoneal space and the difficulty of converting a potential space to an actual space. Thus, the authors suggest a single-port robotic system optimized for narrow surgical spaces as a feasible option for DIEP flap harvest by a totally extraperitoneal approach, which has a shorter learning curve.

The Role of Lysobacter antibioticus HS124 on the Control of Fall Webworm (Hyphantria cunea Drury) and Growth Promotion of Canadian Poplar (Populus canadensis Moench) at Saemangeum Reclaimed Land in Korea.

Populus canadensis Moench forests established in Saemangeum-reclaimed land have been invaded by Hyphantria cunea Drury, causing defoliation and stunted growth. This study investigated the biocontrol potential of cuticle degrading chitinase and protease secreted by Lysobacter antibioticus HS124 against H. cunea larvae. In addition, L. antibioticus HS124 was examined for indole-3-acetic acid phytohormone production for plant growth promotion. To determine the larvicidal activity in the laboratory experiments, crude enzymes, bacteria culture, CY medium, and water (control) were sprayed on the larvae reared on natural diet in insect rearing dishes. Treatment with crude enzymes and bacteria culture caused 76.7% and 66.7% larvae mortality, respectively. The larvae cuticle, mainly composed of chitin and proteins, was degraded by cuticle-degrading enzymes, chitinase, and protease in both the bacteria culture and crude enzyme treatments, causing swelling and disintegration of the cuticle. Field application of the bacteria culture was achieved by vehicle-mounted sprayer. Bacterial treatment caused morphological damage on the larvae cuticles and subsequent mortality. Foliar application of the bacteria culture reduced tree defoliation by H. cunea and enhanced growth compared to the control. Especially, L. antibioticus HS124 produced auxins, and increased growth of poplar trees.

Complete mitochondrial genome of the distinct red-eared slider (Trachemys scripta ssp., Testudines: Emydidae) in Korea.

The complete mitochondrial (mt) genome of Trachemys scripta ssp. in Korea was sequenced and characterized. The mt genome is constituted of 37 genes (13 protein-coding genes, 22 transfer RNA genes and 2 ribosomal RNA genes) and a control region. Phylogenetic analysis based on the complete mt genome showed that the unidentified turtle had the mt genome closely related to that of T. s. elegans, though it had distinct morphology compared to T. s. elegans. This study can provide information for biogeographical studies and management plan for invasive species.

Complete mitochondrial genome of the Cumberland slider (Trachemys scripta troostii, Testudienes: Emydidae) in Korea.

The complete mitochondrial (mt) genome of Trachemys scripta troostii was sequenced and was characterized, which comprised 37 genes (13 protein-coding genes, 22 transfer RNAs, and 2 ribosomal RNAs) and a non-coding control region. Phylogenetic analysis based on the full mt genome indicated that T. s. troostii was more closely related to T. scripta from Canada than to T. s. elegans from China or T. s. scripta fom China. This is the first complete mt genome from T. s. troostii, which provides data for further study of phylogeny in Emydidae.

Hydrogel-Based Stamping Technology for Solution-Free Blood Cell Staining.

An accurate microscopical analysis of blood smears requires a reproducible and convenient method of staining. Solution-based staining procedures can be cumbersome. Especially in low- and middle-income countries, the lack of skilled technicians and adequate laboratory facilities, as well as insufficient water and reagent quality, often become confounding factors. To overcome these obstacles, we developed a new cell staining method based on sequential stamping of agarose gel patches that contain eosin, methylene blue/oxidized methylene blue, Azure B, and buffer, respectively. Our method, termed "hydrogel staining", provides a simple, reproducible, solution-free, and inexpensive approach to stain blood cells. We have optimized incubation times to achieve the optimal transfer of dyes to fixed blood cells on a glass slide, with outcomes comparable to conventional solution-based methods for white blood cells and malaria-infected red blood cells. This hydrogel staining method does not require special skills to produce excellent quality stained blood film slides. The new method could enhance the accuracy of microscopical examination of blood smears, especially in resource-limited settings.

Bacillus velezensis CE 100 Inhibits Root Rot Diseases (Phytophthora spp.) and Promotes Growth of Japanese Cypress (Chamaecyparis obtusa Endlicher) Seedlings.

Root rot diseases, caused by phytopathogenic oomycetes, Phytophthora spp. cause devastating losses involving forest seedlings, such as Japanese cypress (Chamaecyparis obtusa Endlicher) in Korea. Plant growth-promoting rhizobacteria (PGPR) are a promising strategy to control root rot diseases and promote growth in seedlings. In this study, the potential of Bacillus velezensis CE 100 in controlling Phytophthora root rot diseases and promoting the growth of C. obtusa seedlings was investigated. B. velezensis CE 100 produced ß-1,3-glucanase and protease enzymes, which degrade the ß-glucan and protein components of phytopathogenic oomycetes cell-wall, causing mycelial growth inhibition of P. boehmeriae, P. cinnamomi, P. drechsleri and P. erythoroseptica by 54.6%, 62.6%, 74.3%, and 73.7%, respectively. The inhibited phytopathogens showed abnormal growth characterized by swelling and deformation of hyphae. B. velezensis CE 100 increased the survival rate of C. obtusa seedlings 2.0-fold and 1.7-fold compared to control, and fertilizer treatment, respectively. Moreover, B. velezensis CE 100 produced indole-3-acetic acid (IAA) up to 183.7 mg/L, resulting in a significant increase in the growth of C. obtusa seedlings compared to control, or chemical fertilizer treatment, respectively. Therefore, this study demonstrates that B. velezensis CE 100 could simultaneously control Phytophthora root rot diseases and enhance growth of C. obtusa seedlings.

Complete mitochondrial genome of the Small Salamander in Korea, Hynobius unisacculus (Anura: Hynobiidae).

The complete mitochondrial (mt) genome of Hynobius unisacculus was sequenced and characterized. The circular mt genome constituted of 37 genes (13 protein-coding genes, 22 transfer RNAs, and 2 ribosomal RNAs) and a non-coding region (NCR). Phylogenetic analysis based on the full mt genome sequences confirmed that H. unisacculus was closely related to Hynobius leechii rather than other Hynobius species. This is the first completed mt genome from H. unisacculus, which provides data for further study of phylogeny in Hynobiidaes.

Effect of induced hyperopia on fall risk and Fourier transformation of postural sway.

BACKGROUND AND PURPOSE: Fall accidents are a social challenge in Korea and elsewhere. Most previous studies have focused on the effects of reduced visual acuity due to myopia on falls and body balance. The objective of this study was to investigate whether uncorrected hyperopia was a major risk factor for falls and to establish whether the risk of falls was absolutely correlated with visual acuity. METHODS: Fifty-one young subjects with a mean age of 22.75 ± 2.13 years were enrolled in this study. To induce hyperopic and myopic refractive errors, spherical lenses of ±1.0-6.0 D (1.0 D stepwise) were used. Under each induced condition, fall risk index and sway power were assessed via Fourier transformation of postural sway using a TETRAX system. RESULTS: The fall risk index for eyes-closed was significantly greater than that of eyes-open with full correction (t = -5.876, p < 0.05). The fall risk index increased significantly from hyperopia induced with -4.0 D lenses (with visual acuity of 0.69 ± 0.32) compared to eyes-open with full correction (F = 3.213, p < 0.05). However, there was no significant change in the induced myopia conditions, despite a drastic decline in decimal visual acuity. Sway power increased significantly in the low-to-medium frequency band derived from the peripheral vestibular system when hyperopia was induced. A significant difference was detected in hyperopia induced with -6.0 D lenses compared to eyes-open with full correction (F = 4.981, p = 0.017). CONCLUSION: An uncorrected hyperopia rather than myopia may increase the risk of falls, although eyes may show normal visual acuity due to the inherent accommodation mechanism. Our findings suggest that the corrected state of refractive errors is more important than the level of visual acuity as the criteria for appropriate visual input, which contributes to stable posture. Therefore, clinicians should consider the refractive condition, especially the characteristics of hyperopia, when analyzing body balance, and appropriate correction of uncorrected hyperopia to prevent falls.

Epigenetic regulation of mammalian Hedgehog signaling to the stroma determines the molecular subtype of bladder cancer.

In bladder, loss of mammalian Sonic Hedgehog (Shh) accompanies progression to invasive urothelial carcinoma, but the molecular mechanisms underlying this cancer-initiating event are poorly defined. Here, we show that loss of Shh results from hypermethylation of the CpG shore of the Shh gene, and that inhibition of DNA methylation increases Shh expression to halt the initiation of murine urothelial carcinoma at the early stage of progression. In full-fledged tumors, pharmacologic augmentation of Hedgehog (Hh) pathway activity impedes tumor growth, and this cancer-restraining effect of Hh signaling is mediated by the stromal response to Shh signals, which stimulates subtype conversion of basal to luminal-like urothelial carcinoma. Our findings thus provide a basis to develop subtype-specific strategies for the management of human bladder cancer.

A universal ultrasound diagnostic system developed to support urology and coloproctological applications.

In this study, we sought to describe a novel imaging apparatus that is lightweight, inexpensive, and highly effective for use in colorectal diagnostic and treatment settings. Typical probes for use in colorectal ultrasonic imaging applications are developed for surgeons to diagnose and stage rectal tumors and image the rectum and anus. Here we outline a new technique and use it for colorectal imaging in an animal. This technique involves use of an ultrasound array module positioned along the axis of rotation such that improved rotation is possible. This module is in the shape of a linear rod with a rotary linear component that allows for emission of focused ultrasonic echo signals from a linear section of the probe. The usability of the transducer and rectal image quality are satisfactory in a porcine model with the technique proposed here, axial/lateral resolution as 0.96/2.24 mm with 6 dB applied through the contour map using the point spread function. When compared to currently available methods, this technique provides superior diagnostic 3D volumetric image quality with reduced acquisition time. Given this, the ultrasound device proposed here may prove a viable and preferable method to those currently available for urology and colorectal imaging applications.

Patient fibroblasts-derived induced neurons demonstrate autonomous neuronal defects in adult-onset Krabbe disease.

Krabbe disease (KD) is an autosomal recessive neurodegenerative disorder caused by defective ß-galactosylceramidase (GALC), a lysosomal enzyme responsible for cleavage of several key substrates including psychosine. Accumulation of psychosine to the cytotoxic levels in KD patients is thought to cause dysfunctions in myelinating glial cells based on a comprehensive study of demyelination in KD. However, recent evidence suggests myelin-independent neuronal death in the murine model of KD, thus indicating defective GALC in neurons as an autonomous mechanism for neuronal cell death in KD. These observations prompted us to generate induced neurons (iNeurons) from two adult-onset KD patients carrying compound heterozygous mutations (p.[K563*];[L634S]) and (p.[N228_S232delinsTP];[G286D]) to determine the direct contribution of autonomous neuronal toxicity to KD. Here we report that directly converted KD iNeurons showed not only diminished GALC activity and increased psychosine levels, as expected, but also neurite fragmentation and abnormal neuritic branching. The lysosomal-associated membrane proteins 1 (LAMP1) was expressed at higher levels than controls, LAMP1-positive vesicles were significantly enlarged and fragmented, and mitochondrial morphology and its function were altered in KD iNeurons. Strikingly, we demonstrated that psychosine was sufficient to induce neurite defects, mitochondrial fragmentation, and lysosomal alterations in iNeurons derived in healthy individuals, thus establishing the causal effect of the cytotoxic GALC substrate in KD and the autonomous neuronal toxicity in KD pathology.

Neuroprotective and antioxidant activities of bamboo salt soy sauce against H2O2-induced oxidative stress in rat cortical neurons.

Bamboo salt (BS) and soy sauce (SS) are traditional foods in Asia, which contain antioxidants that have cytoprotective effects on the body. The majority of SS products contain high levels of common salt, consumption of which has been associated with numerous detrimental effects on the body. However, BS may be considered a healthier substitute to common salt. The present study hypothesized that SS made from BS, known as bamboo salt soy sauce (BSSS), may possess enhanced cytoprotective properties; this was evaluated using a hydrogen peroxide (H2O2)-induced neuronal cell death rat model. Rat neuronal cells were pretreated with various concentrations (0.001, 0.01, 0.1, 1 and 10%) of BSSS, traditional soy sauce (TRSS) and brewed soy sauce (BRSS), and were subsequently exposed to H2O2 (100 µM). The viability of neuronal cells, and the occurrence of DNA fragmentation, was subsequently examined. Pretreatment of neuronal cells with TRSS and BRSS reduced cell viability in a concentration-dependent manner, whereas neuronal cells pretreated with BSSS exhibited increased cell viability, as compared with non-treated neuronal cells. Furthermore, neuronal cells pretreated with 0.01% BSSS exhibited the greatest increase in viability. Exposure of neuronal cells to H2O2 significantly increased the levels of reactive oxygen species (ROS), B-cell lymphoma 2-associated X protein, poly (ADP-ribose), cleaved poly (ADP-ribose) polymerase, cytochrome c, apoptosis-inducing factor, cleaved caspase-9 and cleaved caspase-3, in all cases. Pretreatment of neuronal cells with BSSS significantly reduced the levels of ROS generated by H2O2, and increased the levels of phosphorylated AKT and phosphorylated glycogen synthase kinase-3ß. Furthermore, the observed effects of BSSS could be blocked by administration of 10 µM LY294002, a phosphatidylinositol 3-kinase inhibitor. The results of the present study suggested that BSSS may exert positive neuroprotective effects against H2O2-induced cell death by reducing oxidative stress, enhancing survival signaling, and inhibiting death signals.