Massive parallel sequencing of short tandem repeats in the Korean population.

STR analysis using capillary electrophoresis has been the most widely used method for forensic DNA typing. Recently, massive parallel sequencing (MPS) technique has been emerging as an innovative tool to supplement or replace the conventional CE process. In this study, we evaluated the application of commercial MiSeqFGx™ forensic signature kit (Illumina Inc., San Diego, CA, USA) in the Korean population, including performance comparison with CE-based STR profiling kits. The genotyping results of 209 unrelated random Korean individuals were summarized according to the International Society for Forensic Genetics guideline. The study revealed that 26 novel sequence variations in autosomal STR were newly found that had not been previously reported in other forensic literature. This indicates that MPS may be an effective supplementary tool for forensic DNA typing and the database to increase the discriminatory power of individual identification.

Anti-Melanogenic Activity of Gagunin D, a Highly Oxygenated Diterpenoid from the Marine Sponge Phorbas sp., via Modulating Tyrosinase Expression and Degradation.

Tyrosinase is the rate-limiting enzyme critical for melanin synthesis and controls pigmentation in the skin. The inhibition of tyrosinase is currently the most common approach for the development of skin-whitening cosmetics. Gagunin D (GD), a highly oxygenated diterpenoid isolated from the marine sponge Phorbas sp., has exhibited cytotoxicity toward human leukemia cells. However, the effect of GD on normal cells and the molecular mechanisms remain to be elucidated. In the present study, we identified for the first time the anti-melanogenic activity of GD and its precise underlying mechanisms in mouse melan-a cells. GD significantly inhibited melanin synthesis in the melan-a cells and a reconstructed human skin model. Further analysis revealed that GD suppressed the expression of tyrosinase and increased the rate of tyrosinase degradation. GD also inhibited tyrosinase enzymatic activity. In addition, GD effectively suppressed the expression of proteins associated with melanosome transfer. These findings suggest that GD is a potential candidate for cosmetic formulations due to its multi-functional properties.

Effect of gut microbiota-derived metabolites and extracellular vesicles on neurodegenerative disease in a gut-brain axis chip.

A new perspective suggests that a dynamic bidirectional communication system, often referred to as the microbiome-gut-brain axis, exists among the gut, its microbiome, and the central nervous system (CNS). This system may influence brain health and various brain-related diseases, especially in the realms of neurodevelopmental and neurodegenerative conditions. However, the exact mechanism is not yet understood. Metabolites or extracellular vesicles derived from microbes in the gut have the capacity to traverse the intestinal epithelial barrier or blood-brain barrier, gaining access to the systemic circulation. This phenomenon can initiate the physiological responses that directly or indirectly impact the CNS and its function. However, reliable and controllable tools are required to demonstrate the causal effects of gut microbial-derived substances on neurogenesis and neurodegenerative diseases. The integration of microfluidics enhances scientific research by providing advanced in vitro engineering models. In this study, we investigated the impact of microbe-derived metabolites and exosomes on neurodevelopment and neurodegenerative disorders using human induced pluripotent stem cells (iPSCs)-derived neurons in a gut-brain axis chip. While strain-specific, our findings indicate that both microbial-derived metabolites and exosomes exert the significant effects on neural growth, maturation, and synaptic plasticity. Therefore, our results suggest that metabolites and exosomes derived from microbes hold promise as potential candidates and strategies for addressing neurodevelopmental and neurodegenerative disorders.

Complete genome sequence of human respiratory syncytial virus genotype A with a 72-nucleotide duplication in the attachment protein G gene.

The complete genome sequence of human respiratory syncytial virus genotype A (HRSV-A) with a 72-nucleotide duplication in the C-terminal part of the attachment protein G gene was determined and analyzed. The genome was 15,277 bp in length, and 0.46 to 6.03% variations were identified at the nucleotide level compared with the previously reported complete genome of HRSV-A. Characterization of the genome will improve understanding of the diversity of the HRSV-A major antigens and enable an in-depth analysis of its genetics.

Benefits of Music Intervention on Anxiety, Pain, and Physiologic Response in Adults Undergoing Surgery: A Systematic Review and Meta-analysis.

PURPOSE: Evidence on factors influencing the variations of music's effect on anxiety and pain in surgical patients is unclear. We aimed to elucidate the effects of music intervention on anxiety and pain throughstudy characteristics. METHODS: We conducted a search on the PubMed, CINAHL, Embase, Cochrane, and Web of Science databases from March 7 to April 21, 2022, for randomized controlled trials (RCTs) for the effect of music intervention on anxiety, pain, and physiological responses in surgical patients. We included studies published within the last 10 years. We assessed the risk of bias in the study using the Cochrane risk of bias tool for randomized trials and performed meta-analyses using a random-effects model for all outcomes. We used change-from-baseline scores as summary statistics and computed bias-corrected standardized mean differences (Hedges'g) for anxiety and pain outcomes and mean differences (MD) for blood pressure and heart rate. RESULTS: Of the 454 records retrieved, 30 RCTs involving 2280 participants were found to be eligible. Music intervention was found to be superior to standard care in reducing anxiety (Hedges' g = -1.48, 95% confidence interval: -1.97 to -0.98), pain (Hedges's g = -0.67, -1.11 to -0.23), systolic blood pressure (MD = -4.62, -7.38 to -1.86), and heart rate (MD = -3.37, -6.65 to -0.10) in surgical patients. The impact of music on anxiety and pain relief varied significantly depending on the duration of the intervention. The largest effect was observed in interventions lasting between 30 and 60 minutes, with a decrease in anxiety and pain. CONCLUSIONS: Music intervention is an effective way to reduce anxiety, pain, and physiological responses in surgical patients. Future reviews examining the influence of different types of surgery on the effects of music would add to the body of knowledge in this field. This study has been registered on the International Prospective Register of Systematic Reviews (PROSPERO) under the number CRD42022340203, with a registration date of July 4, 2022.

Characterization and activity-folding relationship of serine protease from Antarctic krill (Euphausia superba).

Euphausia superba (Antarctic krill) serine protease (ESP) was investigated to gain insights into the activity-structural relationship, folding behavior, and regulation of the catalytic function. We purified ESP from the krill muscle and characterized biochemical distinctions via enzyme kinetics. Studies of inhibition kinetics and unfolding in the presence of a serine residue modifier, such as phenylmethanesulfonyl fluoride, were conducted. Structural characterizations were measured by spectrofluorimetry, including 1-anilinonaphthalene-8-sulfonate dye labeling for hydrophobic residues. The computational simulations such as docking and molecular dynamics were finally conducted to detect key residues and folding behaviors in a nano-second range. The kinetic parameters of ESP were measured as KmBANH = 0.97 ± 0.15 mM and kcat/KmBANH = 4.59 s-1/mM. The time-interval kinetics measurements indicated that ESP inactivation was transformed from a monophase to a biphase process to form a thermodynamically stable state. Spectrofluorimetry measurements showed that serine is directly connected to the regional folding of ESP. Several osmolytes such as proline and glycine only partially protected the inactive form of ESP by serine modification. Computational molecular dynamics and docking simulations showed that three serine residues (Ser183, Ser188, and Ser207) and Cys184, Val206, and Gly209 are key residues of catalytic functions. Our study revealed the functional roles of serine residues as key residues of catalytic function at the active site and of the structural conformation as key folding factors, where ESP displays a flexible property of active site pocket compared to the overall structure.Communicated by Ramaswamy H. Sarma.

Oblique paraspinal approach for thoracic disc herniations using tubular retractor with robotic holder: a technical note.

INTRODUCTION: Symptomatic thoracic disc herniations (TDHs) are uncommon and can be surgically treated. Although transthoracic decompression is considered the gold standard, it is associated with significant comorbidities. In particular, approach via a posterior laminectomy has been associated with poor results. Several strategies have been developed for the resection of TDHs without manipulating the spinal cord. We describe a minimally invasive technique by using 3-D navigation and tubular retractors with the aid of a robotic holder via an oblique paraspinal approach. MATERIALS AND METHODS: The 20-mm working tube via an oblique trajectory through the fascia provides a good surgical field for thoracic discectomy through a microscope. We present our first five patients with TDHs operated using this minimally invasive approach. RESULTS: Neurological symptoms were improved postoperatively, and there were no surgical complications. There was no instability or recurrence during the follow-up period. CONCLUSION: The oblique paraspinal approach may offer an alternative surgical option for treating TDHs.

Management of cerebrospinal fluid leakage after anterior decompression for ossification of posterior longitudinal ligament in the thoracic spine: the utilization of a volume-controlled pseudomeningocele.

STUDY DESIGN: Retrospective review OBJECTIVE: To determine the efficacy of management of cerebrospinal fluid (CSF) leakage after the anterior thoracic approach. SUMMARY OF BACKGROUND DATA: CSF leakage after incidental durotomy commonly occurs after anterior thoracic ossification of posterior longitudinal ligament (OPLL) surgery. Pseudomeningocele will invariably form under such circumstances. Among them, uncontrolled CSF leakage with a fistulous condition is problematic. As a solution, we have managed these durotomies with chest drains alone without any CSF drainage by the concept of a "volume-controlled pseudomeningocele." METHODS: Between 2001 and 2009, CSF leakage occurred in 26 patients (37.7%) of the total 69 patients who underwent anterior decompression for thoracic OPLL. In the initial 11 cases, subarachnoid drainage was utilized as an augmentive measure in combination with chest tube drainage in the postoperative period (group A). In the subsequent 15 cases, the durotomy was managed in a similar manner but in the absence of any subarachnoid drainage (group B). Various parameters such as the duration of postoperative hospital stay, clinical outcome score, drainage output, resolution of CSF leakage, complications, and additional surgery performed were analyzed and compared between the 2 groups. A resolution of the CSF leakage grading system was also proposed for the residual pseudomeningocele that formed in each group. RESULTS: There were statistically no significant differences in the outcome parameters between the 2 groups and also in patients with grade I or grade II residual pseudomeningocele of the new grading system. Two complications occurred in group A. No reexploration for persistent CSF leakage was required in both groups. CONCLUSIONS: CSF leakage managed with controlled chest tube drainage can produce a comparable result with those with additional subarachnoid drainage when watertight dural repair is impossible. The concept of controlled pseudomeningocele may be a useful and practical technique for the treatment of CSF leakage after anterior thoracic OPLL surgery.

MRI versus CT for the diagnosis of lumbar spinal stenosis.

OBJECTIVE: The purpose of the study was to compare the effectiveness of CT and MRI in visualizing soft tissues in lumbar spinal stenosis (LSS), and to correlate the images with preoperative symptoms. MATERIALS AND METHODS: A total of 163 patients who had undergone unilateral laminotomy for bilateral decompression to treat LSS at L4-5 were retrospectively analyzed. The narrowed spinal canal area was measured on axial images with CT and MRI, and compared with the acquired dimensions from preoperative Visual Analog Scale (VAS) and Oswestry Disability Index (ODI) scores. RESULTS: The mean compromised spinal canal areas were 75.08 mm(2) on MRI and 63.13 mm(2) on CT, which were significantly different. Mean VAS for back pain was 5.37, and 7.94 for leg pain. Mean ODI was 55.17%. There was no significant correlation noted between clinical parameters and narrowed spinal canal area. CONCLUSION: Spinal canal area was more narrowed on CT than on MRI in axial cuts. This finding can be explained by the superior ability of multidetector CT to discriminate cortical bone from soft tissue such as the ligamentum flavum. Our study highlights the value of CT examination in combination with MRI prior to LSS surgery.

Single-cell transcriptome of the mouse retinal pigment epithelium in response to a low-dose of doxorubicin.

Cellular senescence of the retinal pigment epithelium (RPE) is thought to play an important role in vision-threatening retinal degenerative diseases, such as age-related macular degeneration (AMD). However, the single-cell RNA profiles of control RPE tissue and RPE tissue exhibiting cellular senescence are not well known. We have analyzed the single-cell transcriptomes of control mice and mice with low-dose doxorubicin (Dox)-induced RPE senescence (Dox-RPE). Our results have identified 4 main subpopulations in the control RPE that exhibit heterogeneous biological activities and play roles in ATP synthesis, cell mobility/differentiation, mRNA processing, and catalytic activity. In Dox-RPE mice, cellular senescence mainly occurs in the specific cluster, which has been characterized by catalytic activity in the control RPE. Furthermore, in the Dox-RPE mice, 6 genes that have not previously been associated with senescence also show altered expression in 4 clusters. Our results might serve as a useful reference for the study of control and senescent RPE.

Second-Generation JK-206 Targets the Oncogenic Signal Mediator RHOA in Gastric Cancer.

Ras homologous A (RHOA), a signal mediator and a GTPase, is known to be associated with the progression of gastric cancer (GC), which is the fourth most common cause of death in the world. Previously, we designed pharmacologically optimized inhibitors against RHOA, including JK-136 and JK-139. Based on this previous work, we performed lead optimization and designed novel RHOA inhibitors for greater anti-GC potency. Two of these compounds, JK-206 and JK-312, could successfully inhibit the viability and migration of GC cell lines. Furthermore, using transcriptomic analysis of GC cells treated with JK-206, we revealed that the inhibition of RHOA might be associated with the inhibition of the mitogenic pathway. Therefore, JK-206 treatment for RHOA inhibition may be a new therapeutic strategy for regulating GC proliferation and migration.

Role of oral pentosan polysulfate in Bacillus Calmette-Guérin therapy in patients with non-muscle-invasive bladder cancer.

PURPOSE: Intravesical Bacillus Calmette-Guérin (BCG) instillation, although an important treatment for non-muscle-invasive bladder cancer, exerts local and systemic adverse effects. Pentosan polysulfate (PPS) is a bladder mucosal protective drug that acts by replacing mucus in the glycosaminoglycan layer of the damaged urothelium. We hypothesized that co-administration of oral PPS with BCG instillation would relieve BCG-related adverse effects without affecting its efficacy. MATERIALS AND METHODS: A total of 217 patients receiving BCG instillation were enrolled. They were placed in two groups and analyzed retrospectively: group A (n=122) received BCG instillation only and group B (n=95) received 100 mg of PPS thrice daily during the BCG treatment. RESULTS: After BCG instillation, the rate of BCG-treatment discontinuation owing to adverse effects was 15.6% in group A and 6.3% in group B (p=0.034). The proportion of patients with bacteriuria after BCG was higher in group B; however, no statistical difference was observed (28.7% vs. 41.1%; p=0.057). The proportion of patients with pyuria was significantly higher in group B (81.1% vs. 91.6%; p=0.029). The proportion of patients using antibiotics was significantly higher in group A (73.8% vs. 43.2%; p=0.001). The recurrence rate within 1 year was 29 (23.8%) in group A vs. 19 (20.0%) in group B (p=0.507). Univariate and multivariate analyses showed that antibiotic use had a statistically significant effect on BCG discontinuation. CONCLUSIONS: Oral PPS effectively decreased the discontinuation rate and antibiotic use without affecting the BCG efficacy.

Isolation and molecular characterization of Bacillus thuringiensis subsp. kurstaki toxic to lepidopteran pests Spodoptera spp. and Plutella xylostella.

BACKGROUND: Bacillus thuringiensis (Bt) has been widely used as a biological control agent for lepidopteran pests. However, resistance to Bt is a major concern associated with Spodoptera spp. (Noctuidae) and Plutella xylostella (Plutellidae). For efficient control of Noctuidae and Plutellidae, novel Bt strains which have high toxicity and a broad host range are needed. RESULTS: To develop novel Bt strains as used for bio-insecticides, the Bt IMBL-B9 with high toxicity against Spodoptera exigua, Spodoptera frugiperda and P. xylostella was isolated and characterized. The Bt kurstaki IMBL-B9 strain produced bipyramidal and cuboidal crystals consisting of cry toxins with molecular weights of 130 and 65 kDa, respectively. This strain harbors eight crystal protein genes in total, including cry1Ea and one vegetative insecticidal protein gene. The median lethal concentration (LC50 ) values of IMBL-B9 against S. exigua and S. frugiperda were 21.8- and 19.3-fold lower than those of the Bt kusrstaki strain, and 5.6- and 4.9-fold lower than those of Bt aizawai strain, respectively. To evaluate the insecticidal activity of Cry proteins from IMBL-B9, cry gene-sourced recombinant Bt strains were constructed. These strains have insecticidal activity and synergic action against lepidopteran pests. CONCLUSION: In this study, a novel Bt kurstaki IMBL-B9 strain was isolated and this could be useful for the development of new bio-insecticide or cry gene-based recombinant products as an alternative solution against lepidopterans, including Noctuidae and Plutellidae. © 2022 Society of Chemical Industry.

Transformation of Bacillus thuringiensis plasmid DNA by a new polyethylenimine polymeric nanoparticles method.

Although electroporation technique has been mostly used to transform Bacillus thuringiensis (Bt), this method is not readily applicable to strains other than the one for which it was optimized. Polyethylenimine (PEI) is a golden standard non-viral vector that interacts with plasmids to form compact polymeric nanoparticles (PNPs) via electrostatic interactions. This PNPs system is very attractive because they are easily prepared, able to carry large nucleic acid constructs, and show low toxicity. In this study, PEI/pBTdsSBV-VP1 PNPs were successfully prepared at various N/P ratios which is positively-chargeable polymer amine (N = nitrogen) groups to negatively-charged nucleic acid phosphate (P) groups, and the internalization of the complexes into Bt 4Q7 was confirmed by confocal laser scanning microscopy. The PEI-mediated transformation showed similar efficiency comparable to that of electroporation method, suggesting that the method of PNPs will be an effective alternative for transformation of Bt strains.

Mean platelet volume is the most valuable hematologic parameter in differentiating testicular torsion from epididymitis within the golden time.

Background: We aimed to assess the diagnostic value of hematologic parameters in the differential diagnosis of testicular torsion and epididymitis within and after the golden time. Methods: We retrospectively reviewed the records of 250 patients aged <25 years who were diagnosed with epididymitis (n=119) or testicular torsion (n=131). The characteristics and hematologic parameters of patients in the two groups were analyzed. Receiver operating characteristic (ROC) curves were used to assess the validity of hematologic parameters as differential diagnostic tools with respect to the golden time (defined as 6 h of symptom duration). Further, we evaluated the predictive factors associated with orchiectomy in patients with testicular torsion. Results: The mean patient age was 14.4 years. Among patients with testicular torsion, 91.40% (53 of 58) underwent detorsion and orchiopexy within the golden time, whereas only 27.40% (20 of 73) of the affected testes were preserved after the golden time. Within the golden time, mean platelet volume (MPV) seemed to be the most valuable hematologic parameter [area under the curve (AUC) 0.855, 95% confidence interval (CI): 0.778-0.932]. In a multivariate analysis, symptom duration (symptoms beyond the golden time) was associated with orchiectomy in patients with testicular torsion. Conclusions: MPV showed the greatest hematologic value in the early stage of testicular torsion and epididymitis, suggesting its potential use for the differential diagnosis of these two conditions within the golden time.

Dural tears in percutaneous endoscopic lumbar discectomy.

The objective of the study was to demonstrate the clinical characteristics of dural tears during percutaneous endoscopic lumbar discectomy (PELD) and to discuss how to prevent this unintended complication. The study used data from 816 consecutive patients who underwent PELD between 2003 and 2007. A total of nine patients (1.1%) experienced symptomatic dural tears. The clinical outcomes were evaluated using the visual analogue scale (VAS), the Oswestry disability index (ODI), and modified MacNab criteria. Intractable radicular pain was the most common symptom, while classical manifestations, such as CSF leakage or wound swelling, were rare. In three of the nine cases, the dural tears were detected intraoperatively, while the remaining six cases were not recognized during the procedure. Among the unrecognized patients, two patients were found with nerve root herniation causing profound neurological deficits. All patients were managed by secondary open repair surgeries. The mean follow-up period was 30.8 months. The mean VAS of radicular leg pain improved from 8.3 to 2.6, and that of back pain improved from 4.1 to 2.6. The mean ODI improved from 69.6 to 29.2%. The final outcomes were excellent in one, good in five, fair in one, and poor in two patients. As application of the endoscopic procedure has been broadened to more complex cases, the risk of dural tears may increase. Unrecognized dural tear with nerve root herniation may cause permanent neurological sequelae. Accurate information and proper technical considerations are essential to prevent this unpredictable complication.

Cervical cord compression presenting with sciatica-like leg pain.

Sciatica-like leg pain can be the main presenting symptom in patients with cervical cord compression. It is a false localizing presentation, which may lead to missed or delayed diagnosis, resulting in the wrong plan of management, especially in the presence of concurrent lumbar lesions. Medical history, physical findings and the results of imaging studies were reviewed in two cases of cervical cord compressions, which presented with sciatica-like leg pain. There was multi-level cervical spondylosis with cord compression in the first patient and the second patient had two levels of cervical disc herniation with cord compression. In both cases, there were co-existing lumbar lesions, which could be responsible for the presentation of the leg pain. Cervical blocks were diagnostic in identifying the level responsible for the leg pain and it was confirmed so after cervical decompressive surgery in both cases, which brought significant pain relief. Funicular leg pain is a rare presentation of cervical cord compression. It is a referred pain due to the irritation of the ascending spinothalamic tract. Cervical blocks were successful in identifying the cause of funicular pain in our cases and this may pave the way for further studies to establish the role of cervical blocks as a diagnostic tool for funicular pain caused by cord compression.

Targeting senescent retinal pigment epithelial cells facilitates retinal regeneration in mouse models of age-related macular degeneration.

Although age-related macular degeneration (AMD) is a multifactorial disorder with angiogenic, immune, and inflammatory components, the most common clinical treatment strategies are antiangiogenic therapies. However, these strategies are only applicable to neovascular AMD, which accounts for less than 20% of all AMD cases, and there are no FDA-approved drugs for the treatment of dry AMD, which accounts for ~ 80% of AMD cases. Here, we report that the elimination of senescent cells is a potential novel therapeutic approach for the treatment of all types of AMD. We identified senescent retinal pigment epithelium (RPE) cells in animal models of AMD and determined their contributions to retinal degeneration. We further confirmed that the clearance of senescent RPE cells with the MDM2-p53 inhibitor Nutlin-3a ameliorated retinal degeneration. These findings provide new insights into the use of senescent cells as a therapeutic target for the treatment of AMD.

Genetic Differences between Physical Injury Patients With and Without Post-traumatic Syndrome: Focus on Secondary Findings and Potential Variants Revealed by Whole Exome Sequencing.

Objective: Sudden traumatic physical injuries often cause psychological distress, which may be associated with chronic disability. Although considerable effort has been expended to identify genetic predictors of post-traumatic stress disorder (PTSD) after traumatic events, genetic predictors of psychological distress in response to severe physical injuries have been yet to be elucidated using whole exome sequencing (WES). Here, the genetic architecture of post-traumatic syndrome (PTS), which encompasses a broad range of psychiatric disorders after traumatic events including depression, anxiety disorder, acute stress disorder, and PTSD, was explored using WES in severely physically injured patients, focusing on secondary findings and potential PTS-related variants. Methods: In total, 141 severely physically injured patients were consecutively recruited, and PTS was evaluated within 1 month of the injury. Secondary findings were analyzed according to PTS status. To identify PTS-related variants, genome-wide association analyses and the optimal sequencing kernel association test were performed. Results: Of the 141 patients, 88 (62%) experienced PTS. There were 108 disease-causing variants in severely physically injured patients. As secondary findings, the stress- and inflammation-related signaling pathways were enriched in the PTS patients, while the glucose metabolism pathway was enriched in those without PTS. However, no significant PTS-related variants were identified. Conclusion: Our findings suggest that genetic alterations in stress and inflammatory pathways might increase the likelihood of PTS immediately after severe physical injury. Future studies with larger samples and longitudinal designs are needed.

Gene Expression Profile in Similar Tissues Using Transcriptome Sequencing Data of Whole-Body Horse Skeletal Muscle.

Horses have been studied for exercise function rather than food production, unlike most livestock. Therefore, the role and characteristics of tissue landscapes are critically understudied, except for certain muscles used in exercise-related studies. In the present study, we compared RNA-Seq data from 18 Jeju horse skeletal muscles to identify differentially expressed genes (DEGs) between tissues that have similar functions and to characterize these differences. We identified DEGs between different muscles using pairwise differential expression (DE) analyses of tissue transcriptome expression data and classified the samples using the expression values of those genes. Each tissue was largely classified into two groups and their subgroups by k-means clustering, and the DEGs identified in comparison between each group were analyzed by functional/pathway level using gene set enrichment analysis and gene level, confirming the expression of significant genes. As a result of the analysis, the differences in metabolic properties like glycolysis, oxidative phosphorylation, and exercise adaptation of the groups were detected. The results demonstrated that the biochemical and anatomical features of a wide range of muscle tissues in horses could be determined through transcriptome expression analysis, and provided proof-of-concept data demonstrating that RNA-Seq analysis can be used to classify and study in-depth differences between tissues with similar properties.