An optimized cyclophosphamide-treated mouse model of Mycobacterium abscessus pulmonary infection.

Mycobacterium abscessus pulmonary infections are increasingly problematic, especially for immunocompromised individuals and those with underlying lung conditions. Currently, there is no reliable standardized treatment, underscoring the need for improved preclinical drug testing. We present a simplified immunosuppressed mouse model using only four injections of cyclophosphamide, which allows for sustained M. abscessus lung burden for up to 16 days. This model proved effective for antibiotic efficacy evaluation, as demonstrated with imipenem or amikacin.

Single-Cell Oral Delivery Platform for Enhanced Acid Resistance and Intestinal Adhesion.

Oral delivery of cells, such as probiotics and vaccines, has proved to be inefficient since cells are generally damaged in an acidic stomach prior to arrival at the intestine to exert their health benefits. In addition, short retention in the intestine is another obstacle which affects inefficiency. To overcome these obstacles, a cell-in-shell structure was designed with pH-responsive and mucoadhesive properties. The pH-responsive shell consisting of three cationic layers of chitosan and three anionic layers of trans-cinnamic acid (t-CA) was made via layer-by-layer (LbL) assembly. t-CA layers are hydrophobic and impermeable to protons in acid, thus enhancing cell gastric resistance in the stomach, while chitosan layers endow strong interaction between the cell surface and the mucosal wall which facilitates cell mucoadhesion in the intestine. Two model cells, probiotic L. rhamnosus GG and dead Streptococcus iniae, which serve as inactivated whole-cell vaccine were chosen to test the design. Increased survival and retention during oral administration were observed for coated cells as compared with naked cells. Partial removal of the coating (20-60% removal) after acid treatment indicates that the coated vaccine can expose its surface immunogenic protein after passage through the stomach, thus facilitating vaccine immune stimulation in the intestine. As a smart oral delivery platform, this design can be extended to various macromolecules, thus providing a promising strategy to formulate oral macromolecules in the prevention and treatment of diseases at a cellular level.

Nontoxic, Biodegradable Hyperbranched Poly(ß-amino ester)s for Efficient siRNA Delivery and Gene Silencing.

RNA interference (RNAi)-mediated gene silencing is a promising therapeutic approach to treat various diseases, but safe and efficient delivery remains a major challenge to its clinical application. Non-viral gene vectors, such as poly(ß-amino esters) (pBAEs), have emerged as a potential candidate due to their biodegradability, low toxicity profile, ease of synthesis, and high gene transfection efficiency for both DNA and siRNA delivery. However, achieving significant gene silencing using pBAEs often requires a large amount of polymer carrier (with polymer/siRNA weight ratio >100) or high siRNA dose (>100 nM), which might potentially exacerbate toxicity concerns during delivery. To overcome these barriers, we designed and optimized a series of hyperbranched pBAEs capable of efficiently condensing siRNA and achieving excellent silencing efficiency at a lower polymer/siRNA weight ratio (w/w) and siRNA dose. Through modulation of monomer combinations and branching density, we identified the top-performing hyperbranched pBAEs, named as h(A2B3)-1, which possess good siRNA condensation ability, low cytotoxicity, and high cellular uptake efficiency. Compared with Lipofectamine 2000, h(A2B3)-1 achieved lower cytotoxicity and higher siRNA silencing efficiency in HeLa cells at a polymer/siRNA weight ratio of 30 and 30 nM siRNA dose. Notably, h(A2B3)-1 enhanced the gene uptake in primary neural cells and effectively silenced the target gene in hard-to-transfect primary cortical neurons and oligodendrocyte progenitor cells, with gene knockdown efficiencies of 34.8 and 53.4% respectively. By incorporating a bioreducible disulfide compartment into the polymer backbone, the cytocompatibility of the h(A2B3)-1 was greatly enhanced while maintaining their good transfection efficiency. Together, the low cytotoxicity and high siRNA transfection efficiency of hyperbranched h(A2B3)-1 in this study demonstrated their great potential as a non-viral gene vector for efficient siRNA delivery and RNAi-mediated gene silencing. This provides valuable insight into the future development of safe and efficient non-viral siRNA delivery systems as well as their translation into clinical applications.

Activation of ChAT+ neuron in dorsal motor vagus (DMV) increases blood glucose through the regulation of hepatic gene expression in mice.

The brain and peripheral organs communicate through hormones and neural connections. Proper communication is required to maintain normal whole-body energy homeostasis. In addition to endocrine system, from the perspective of neural connections for metabolic homeostasis, the role of the sympathetic nervous system has been extensively studied, but understanding of the parasympathetic nervous system is limited. The liver plays a central role in glucose and lipid metabolism. This study aimed to clarify the innervation of parasympathetic nervous system in the liver and its functional roles in metabolic homeostasis. The liver-specific parasympathetic nervous system innervation (PNS) was shown by tissue clearing, immunofluorescence and transgenic mice at the three-dimensional histological level. The parasympathetic efferent signals were manipulated using a chemogenetic technique and the activation of ChAT+ parasympathetic neurons in dorsal motor vagus (DMV) results in the increased blood glucose through the elevated hepatic gluconeogenic and lipogenic gene expression in the liver. Thus, our study showed the evidence of ChAT+ parasympathetic neurons in the liver and its role for hepatic parasympathetic nervous signaling in glucose homeostasis through the regulation of hepatic gene expression.

Hydrogel dressings with intrinsic antibiofilm and antioxidative dual functionalities accelerate infected diabetic wound healing.

Chronic wounds are often infected with biofilm bacteria and characterized by high oxidative stress. Current dressings that promote chronic wound healing either require additional processes such as photothermal irradiation or leave behind gross amounts of undesirable residues. We report a dual-functionality hydrogel dressing with intrinsic antibiofilm and antioxidative properties that are synergistic and low-leaching. The hydrogel is a crosslinked network with tethered antibacterial cationic polyimidazolium and antioxidative N-acetylcysteine. In a murine diabetic wound model, the hydrogel accelerates the closure of wounds infected with methicillin-resistant Staphylococcus aureus or carbapenem-resistant Pseudomonas aeruginosa biofilm. Furthermore, a three-dimensional ex vivo human skin equivalent model shows that N-acetylcysteine promotes the keratinocyte differentiation and accelerates the re-epithelialization process. Our hydrogel dressing can be made into different formats for the healing of both flat and deep infected chronic wounds without contamination of the wound or needing other modalities such as photothermal irradiation.

Clinical Aspects of Transient Myopia after Blunt Eye Trauma

Purpose@#To investigate the clinical aspects of transient myopia after blunt eye trauma. @*Methods@#The medical records of 32 patients treated at our hospital for from 2020 to 2022 traumatic microhyphema were analyzed retrospectively. The visual acuity, intraocular pressure, anterior chamber cells, anterior chamber depth, axial length, lens thickness, and refractive index were analyzed immediately after injury and after treatment. Patients with additional diseases such as vitreous and retinal hemorrhage, traumatic retinal detachment, iridodialysis, or orbital fracture were excluded. Correlations between the myopic changes and the amount of myopic refractive change caused by trauma were analyzed. @*Results@#The 32 patients included 24 (75%) males and 8 (25%) females. The average myopic change was -3.03 ± 0.92 (range -1.00 to -5.25) diopters (D). The average change in anterior chamber depth due to trauma was -0.22 ± 0.11 mm, the change in lens diameter was 0.20 ± 0.09 mm, and the change in axial length was -0.07 ± 0.05 mm. The average change in refractive index due to ciliary spasm was -1.63 ± 0.65 D. The correlations between the change in refractive index and anterior chamber depth (r = 0.475, p = 0.008), traumatic ciliary spasm (r = 0.649, p < 0.001), and lens thickness (r = -0.184, p = 0.330) were determined. @*Conclusions@#Factors such as ciliary spasm, change in the anterior chamber depth, anterior shift of the lens-iris diaphragm, and increased lens thickness affect the myopic changes caused by trauma. Of these, a shallower anterior chamber depth and ciliary spasm are highly correlated with the transient myopic changes.

Pediatric Eosinophilic Granuloma Associated With Delayed Epidural Hematoma Following on Seizure: A Case Report

Eosinophilic granuloma (EG), a subtype of Langerhans cell histiocytosis (LCH), the monostotic form, is a rare condition characterized by a solitary bone lesion. It is even more unusual for this condition to be accompanied by an epidural hematoma (EDH). This case is unique in that it is the first to involve delayed EDH following a seizure. We describe a remarkable example of EG accompanied by an EDH and consider the rarity of this comorbidity. A 32-month-old boy developed a rapidly growing skull mass following a minor head injury. During surgical preparation for a biopsy, the patient experienced a single convulsion. Imaging following the seizure revealed an EDH in the vicinity of the mass. The mass was excised and confirmed to be an EG, but with positive margins. The patient underwent chemotherapy after systemic skeletal evaluation, in accordance with the LCH III protocol established by the Histiocytosis Society. EG is a rare neoplasm that typically presents as a painless growth on the skull that gradually enlarges over time. The correlation between EG and EDH is exceedingly uncommon, with only a few documented cases. This case study underscores the significance of considering EG in the differential diagnosis of an expanding cranium mass, even when associated with EDH. Prompt diagnosis and treatment can prevent serious complications and improve patient outcomes.

Treatment for appendicitis in cancer patients on chemotherapy: a retrospective cohort study

Purpose@#Whether to perform surgery or conservatively manage appendicitis in immunosuppressed patients is a concern for clinicians. This study aimed to compare the outcomes of these 2 treatment options for appendicitis in patients with cancer undergoing chemotherapy. @*Methods@#This retrospective study included 206 patients with cancer who were diagnosed with acute appendicitis between August 2001 and December 2021. Among them, patients who received chemotherapy within 1 month were divided into surgical and conservative groups. We evaluated the outcomes, including treatment success within 1 year, 1-year recurrence, and the number of days from the diagnosis of appendicitis to chemotherapy restart, between the 2 groups. @*Results@#Among the 206 patients with cancer who were diagnosed with acute appendicitis, 78 received chemotherapy within 1 month. The patients were divided into surgery (n = 63) and conservative (n = 15) groups. In the surgery group, the duration of antibiotic therapy (7.0 days vs. 16.0 days, P < 0.001) and length of hospital stay (8.0 days vs. 27.5 days, P = 0.002) were significantly shorter than conservative groups. The duration from the diagnosis of appendicitis to the restart of chemotherapy was shorter in the surgery group (20.8 ± 15.1 days vs. 35.2 ± 28.2 days, P = 0.028). The treatment success rate within 1 year was higher in the surgery group (100% vs. 33.3%, P < 0.001). @*Conclusion@#Surgical treatment showed a significantly higher success rate than conservative treatment for appendicitis in patients less than 1 month after chemotherapy. Further prospective studies will be needed to clinically determine treatment options.

Guidelines for Manufacturing and Application of Organoids: Brain

This study offers a comprehensive overview of brain organoids for researchers. It combines expert opinions with technical summaries on organoid definitions, characteristics, culture methods, and quality control. This approach aims to enhance the utilization of brain organoids in research. Brain organoids, as three-dimensional human cell models mimicking the nervous system, hold immense promise for studying the human brain. They offer advantages over traditional methods, replicating anatomical structures, physiological features, and complex neuronal networks. Additionally, brain organoids can model nervous system development and interactions between cell types and the microenvironment. By providing a foundation for utilizing the most human-relevant tissue models, this work empowers researchers to overcome limitations of two-dimensional cultures and conduct advanced disease modeling research.

Long-term clinical outcomes after high and low ligations with lymph node dissection around the root of the inferior mesenteric artery in patients with rectal cancer

Purpose@#This study aimed to evaluate the long-term clinical outcomes based on the ligation level of the inferior mesenteric artery (IMA) in patients with rectal cancer. @*Methods@#This was a retrospective analysis of a prospectively collected database that included all patients who underwent elective low anterior resection for rectal cancer between January 2013 and December 2019. The clinical outcomes included oncological outcomes, postoperative complications, and functional outcomes. The oncological outcomes included overall survival (OS) and relapse-free survival (RFS). The functional outcomes, including defecatory and urogenital functions, were analyzed using the Fecal Incontinence Severity Index, International Prostate Symptom Score, and International Index of Erectile Function questionnaires. @*Results@#In total, 545 patients were included in the analysis. Of these, 244 patients underwent high ligation (HL), whereas 301 underwent low ligation (LL). The tumor size was larger in the HL group than in the LL group. The number of harvested lymph nodes (LNs) was higher in the HL group than in the LL group. There were no significant differences in complication rates and recurrence patterns between the groups. There were no significant differences in 5-year RFS and OS between the groups. Cox regression analysis revealed that the ligation level (HL vs. LL) was not a significant risk factor for oncological outcomes. Regarding functional outcomes, the LL group showed a significant recovery in defecatory function 1 year postoperatively compared with the HL group. @*Conclusion@#LL with LNs dissection around the root of the IMA might not affect the oncologic outcomes comparing to HL; however, it has minimal benefit for defecatory function.

Drug Retention Rate and Factors Associated with Discontinuation of Interleukin-17 Inhibitors in Patients with Axial Spondyloarthritis

Purpose@#To assess the drug retention rate of interleukin-17 inhibitors (IL-17is) over long-term observation in patients with axial spondyloarthritis (axSpA) in whom treatment with tumor necrosis factor inhibitors (TNFis) failed and to determine baseline factors associated with discontinuation of IL-17is. @*Materials and Methods@#This retrospective cohort study included 68 patients with axSpA started on IL-17is after an inadequate response or intolerance to ≥1 TNFis. Drug retention rates at 1, 2, and 3 years were assessed. Baseline (i.e., at initiation of IL-17is) factors associated with discontinuation of IL-17is were evaluated using multivariable Cox proportional hazard regression analysis. @*Results@#Over 1933.9 person-months of observation in 68 patients, discontinuation of IL-17is occurred in 27 (39.7%) patients. Twenty (29.4%) patients discontinued IL-17is because of ineffectiveness, and 7 (10.3%) patients discontinued IL-17is because of adverse events. The 1-year, 2-year, and 3-year drug retention rates for IL-17is were 71.9%, 66.5%, and 62.0%, respectively. Current smoking was associated with a higher risk of IL-17is discontinuation [adjusted hazard ratio (HR)=2.256, 95% confidence interval (CI)=1.053–4.831, p=0.036], while previous use of ≥3 TNFis (vs. 1) was significantly associated with a lower risk of IL-17is discontinuation (adjusted HR=0.223, 95% CI=0.051–0.969, p=0.045). @*Conclusion@#In patients with axSpA in whom TNFis failed, the long-term drug retention rate of IL-17is appears to be acceptable, with a 3-year drug retention rate of approximately 60%. Current smoking was associated with a higher risk of discontinuing IL-17is, whereas previous use of ≥3 TNFis was associated with a lower risk of discontinuing IL-17is.

Comparison between Placement of an Amniotic Membrane Contact Lens and Temporary Amniotic Membrane Transplantation

Purpose@#We compared the clinical utilities of an amniotic contact lens and temporary amniotic membrane transplantation in patients with persistent corneal epithelial defects. @*Methods@#Patients diagnosed with persistent corneal epithelial defects from November 2019 to July 2022 were randomly assigned to either lens placement for 2 weeks or temporary transplantation for 1 week, and the outcomes were retrospectively analyzed. The best-corrected visual acuity, corneal sensation, tear film break-up time, and corneal opacity were measured before and after the interventions. The treatment success rates, recurrences, and other complications were analyzed during the follow-up period. @*Results@#We placed lenses in 32 patients (32 eyes) and 20 patients (20 eyes) underwent transplantation. In both groups, significant differences in best-corrected visual acuity, corneal sensation, tear film break-up time, and corneal opacity were apparent after intervention vs. before. None of the parameters showed any significant differences in terms of their rates of change. @*Conclusions@#In patients with persistent corneal epithelial defects, amniotic contact lens placement is an effective outpatient procedure and is not inferior to temporary amniotic membrane transplantation.

Transition Substitution of Desired Bases in Human Pluripotent Stem Cells with Base Editors: A Step-by-Step Guide

The recent advances in human pluripotent stem cells (hPSCs) enable to precisely edit the desired bases in hPSCs to be used for the establishment of isogenic disease models and autologous ex vivo cell therapy. The knock-in approach based on the homologous directed repair with Cas9 endonuclease, causing DNA double-strand breaks (DSBs), produces not only insertion and deletion (indel) mutations but also deleterious large deletions. On the contrary, due to the lack of Cas9 endonuclease activity, base editors (BEs) such as adenine base editor (ABE) and cytosine base editor (CBE) allow precise base substitution by conjugated deaminase activity, free from DSB formation. Despite the limitation of BEs in transition substitution, precise base editing by BEs with no massive off-targets is suggested to be a prospective alternative in hPSCs for clinical applications. Considering the unique cellular characteristics of hPSCs, a few points should be considered. Herein, we describe an updated and optimized protocol for base editing in hPSCs. We also describe an improved methodology for CBE-based C to T substitutions, which are generally lower than A to G substitutions in hPSCs.

Migrated coil and damaged stent removal during coil embolization, using an additional, retrievable stent: A case report

One of the common complications that can occur during coil embolization of cerebral aneurysms, is migration of coil lump alone. The removal of these migrated coils has been reported on a few occasions. On the other hand, rare complications would include the migration of the coil with subsequent stent dislocation. Currently, there is no standardized method to correct the complications of stent dislocation, and very few instances of this complication have been reported previously. In this report, we introduce a case of coil migration combined with stent dislocation. This occurred during coil embolization of an unruptured aneurysm of the distal, left internal carotid artery for a 52-year old woman. We retrieved both the damaged stent and migrated coil using another retrievable stent successfully with no more further complications. In the present report, we describe in detail how we corrected the complication successfully stent, and we discuss why this rescue maneuver is reasonable option for the complication mentioned above.

Long-term Clinical Course of Amniotic Membrane after Permanent Amniotic Membrane Transplantation

Purpose@#To investigate the long-term clinical outcomes of transplanted amniotic membrane in patients who underwent permanent amniotic membrane transplantation (P-AMT) on cornea. @*Methods@#From April 2014 to March 2021, medical records and digital photographs of 68 patients (72 eyes) who underwent P-AMT were analyzed retrospectively. The duration of complete re-epithelization of cornea and wearing therapeutic contact lense (T-lens) after surgery were investigated, the size of preserved amniotic membrane (AM) excluding the melted portion was analyzed using the Image J program every year up to 6 years after surgery, and it was compared by dividing into a single-layer group and a double-layer group. In addition, when the AM melts, the cause, time, and related factors of melting were analyzed. @*Results@#The average duration to complete re-epithelialization after surgery was 12.2 ± 11.0 days, and T-lens were worn on average up to 8.7 ± 8.5 months after surgery. In total group, the average ratio of preserved AM annually up to 6 years after surgery was 94.9%, 94.3%, 97.8%, 96.4%, 95.8%, 91.6% respectively, and there was no significant difference between the single-layer group and the double-layer group. AM melting appeared in 38.9% (28 eyes) of the total group, and melting was first observed on average 16.4 ± 17.2 months after surgery. The formation and rupture of bullae was the most common cause of melting at 71.4% (20 eyes), and there were no statistically significant related factors involved in the formation of bullae. @*Conclusions@#In the case of P-AMT performed in various diseases of the cornea, the transplanted AM is partially melt but considered to be well preserved up to 6 years after surgery to form a stable ocular surface.

3D printed surveyed restoration and metal framework in removable dentures: A case report / 대한치과보철학회지

Computer-aided design–computer-aided manufacturing technology has been widely used in the manufacture of fixed prostheses including implants, but in the case of removable dentures, the analog method is still being used due to the errors such as a lack of fusion and over-fusion in selective laser meting process. With the recent development of CAD software, virtual surveying and framework design are made possible, and the designed file can be manufactured by milling or 3D printing. It replace the analog method of waxing and denture curing process and also can reduce the production time and cost. Therefore, this case is reported because good clinical results were obtained by digitally surveying on CAD software to produce a surveyed metal restoration and framework on maxillary and mandibular removable dentures.

Digital immediate implantation and aesthetic immediate loading on maxillary incisor displaced due to root fracture: a case report

To obtain better esthetic results when immediately placing a dental implant, the soft tissue surrounding the implant must be conditioned during healing of the extraction socket. To this end, the emergence profile can be customized through immediate restoration of the provisional prosthesis, and good clinical results can be obtained at the time of definitive restoration in the future, resulting in high patient satisfaction. In this case, horizontal root fracture occurred after trauma to both maxillary central incisors.Immediate implant placement and loading was planned considering aesthetics and alveolar bone condition. By taking an impression using a digital intraoral scanner, a digital diagnostic wax-up was performed to make a more aesthetic prosthesis without applying external force to the traumatized teeth. Based on this, the ideal placement location was determined and immediate implant placement was performed using a 3D printed surgical guide. The provisional prosthesis was restored 5 days after placement, and the definitive zirconia crown was restored through soft tissue conditioning and customization using the shape of the provisional prosthesis for 3 months.

Improvement of peri-implant complications through customized prosthesis restoration allowing soft tissue space: a case report

The concept of ‘Implant Supracrestal Complex (ISC)’ was introduced as a way to determine the impact of implant prosthetic design form on both short-term clinical outcomes and the long-term prognosis of bone and soft tissues around the implant.Implant-prosthesis-abutment complex design forms, such as implant-abutment design, junction, and location, can have important implications for the stable and healthy long-term maintenance of the tissues around the implant. In this case, a customized concave abutment and a prosthesis with an emergence angle of about 30 degrees were restored to a patient suffering from chronic soft tissue inflammation and pain after restoration of an implant prosthesis. It was confirmed that the new prosthesis improved complications by allowing sufficient bone and soft tissue space, achieved appropriate soft tissue sealing, and maintained the longterm stability of the implant.

Using treatment denture and digital technology in patient with unstable mandibular movement: a case report

Long-term use of inappropriate prosthesis often results in habitual closure or irregular mandibular movements. In that case, guide to the stable centric position is difficult. Therefore, by using a treatment denture, the muscles and TMJ should be stabilized and the jaw relation should be acquired with the treatment position. Compared to the conventional method, digital technology in fabrica-tion complete denture has the advantage of reproducing a stable tooth arrangement in cases of difficult tooth alignment, minimiz-ing laboratory errors and reducing denture fabrication time. Therefore, in this case, the final denture was fabricated by digitally re-producing the stable treatment position, vertical dimension, and lip support with a treatment denture, and satisfactory results wereobtained.

Fabrication of CAD-CAM complete denture using existing provisional denture and digital facebow transfer / 대한치과보철학회지

In this case, a CAD-CAM complete denture that can be easily remanufactured and reduced the number of visits was decided in consideration of the fact that it was difficult to visit the dental hospital and many existing dentures were lost because the patient was inpatient in a nursing hospital. In order to reflect the shape, vertical dimension, and maxillomandibular relationship of the existing provisional dentures adapted by the patient to the fabrication of the final denture, the existing provisional dentures were scanned and closed mouth impression was taken with the printed individual tray. After digital facebow transfer using UTS CAD and arranging artificial teeth in CAD software, the maxillary and mandibular complete dentures were fabricated by milling the denture base and artificial teeth at once with the monolithic disc.