Nanoscale single-vesicle analysis: High-throughput approaches through AI-enhanced super-resolution image analysis.

The analysis of membrane vesicles at the nanoscale level is crucial for advancing the understanding of intercellular communication and its implications for health and disease. Despite their significance, the nanoscale analysis of vesicles at the single particle level faces challenges owing to their small size and the complexity of biological fluids. This new vesicle analysis tool leverages the single-molecule sensitivity of super-resolution microscopy (SRM) and the high-throughput analysis capability of deep-learning algorithms. By comparing classical clustering methods (k-means, DBSCAN, and SR-Tesseler) with deep-learning-based approaches (YOLO, DETR, Deformable DETR, and Faster R-CNN) for the analysis of super-resolution fluorescence images of exosomes, we identified the deep-learning algorithm, Deformable DETR, as the most effective. It showed superior accuracy and a reduced processing time for detecting individual vesicles from SRM images. Our findings demonstrate that image-based deep-learning-enhanced methods from SRM images significantly outperform traditional coordinate-based clustering techniques in identifying individual vesicles and resolving the challenges related to misidentification and computational demands. Moreover, the application of the combined Deformable DETR and ConvNeXt-S algorithms to differently labeled exosomes revealed its capability to differentiate between them, indicating its potential to dissect the heterogeneity of vesicle populations. This breakthrough in vesicle analysis suggests a paradigm shift towards the integration of AI into super-resolution imaging, which is promising for unlocking new frontiers in vesicle biology, disease diagnostics, and the development of vesicle-based therapeutics.

Factors influencing the overall satisfaction of teachers participating in a nationwide school-based smoking prevention program in Korea.

INTRODUCTION: Numerous studies of school-based smoking prevention programs (SSPPs) exist; however, most have been conducted from the students' perspective, and insufficient research has explored teachers' perceptions. Our study aimed to identify factors affecting overall satisfaction and operational status from the perspective of teachers participating in the SSPP. METHODS: This is a cross-sectional study analyzing data from a survey regarding the operation of an SSPP conducted by the Korea Health Promotion Institute in 2022. The study sample comprised 669 teachers involved in the SSPP: 215 from elementary schools, 212 from middle schools, and 242 from high schools. To identify factors influencing teachers' overall satisfaction, independent variables were categorized into three types of factors: personal, school, and teacher perceptions. Multiple linear regression analysis was performed for each factor to test the independent association. RESULTS: For elementary school teachers, as the necessity of smoking prevention and cessation education (ß=0.292; 95% CI: 0.182-0.382) increased, the overall satisfaction with the operation of the SSPP significantly improved. Similarly, for middle school teachers, as the necessity of smoking prevention and cessation education (ß=0.231; 95% CI: 0.104-0.336) increased, the overall satisfaction significantly improved. Conversely, for high school teachers, the effectiveness (ß=0.347; 95% CI: 0.184-0.520) was the variable that significantly improved overall satisfaction with the SSPP operation. The variables affecting teacher satisfaction across all school levels were the necessity of smoking prevention and cessation education, the effectiveness of the SSPP, and its impact on smoking cessation among smoking students, all of which belonged to the teacher perceptions factor. CONCLUSIONS: Smoking education in schools requires teachers to play a crucial role. Among participating teachers, overall satisfaction with SSPP operations and the influencing factors differed according to school level, highlighting the importance of careful consideration to establish a more effective operational environment tailored to each school level.

How Extracellular Reactive Oxygen Species Reach Their Intracellular Targets in Plants.

Reactive oxygen species (ROS) serve as secondary messengers that regulate various developmental and signal transduction processes, with ROS primarily generated by NADPH OXIDASEs (referred to as RESPIRATORY BURST OXIDASE HOMOLOGs [RBOHs] in plants). However, the types and locations of ROS produced by RBOHs are different from those expected to mediate intracellular signaling. RBOHs produce O2•- rather than H2O2 which is relatively long-lived and able to diffuse through membranes, and this production occurs outside the cell instead of in the cytoplasm, where signaling cascades occur. A widely accepted model explaining this discrepancy proposes that RBOH-produced extracellular O2•- is converted to H2O2 by superoxide dismutase and then imported by aquaporins to reach its cytoplasmic targets. However, this model does not explain how the specificity of ROS targeting is ensured while minimizing unnecessary damage during the bulk translocation of extracellular ROS (eROS). An increasing number of studies have provided clues about eROS action mechanisms, revealing various mechanisms for eROS perception in the apoplast, crosstalk between eROS and reactive nitrogen species, and the contribution of intracellular organelles to cytoplasmic ROS bursts. In this review, we summarize these recent advances, highlight the mechanisms underlying eROS action, and provide an overview of the routes by which eROS-induced changes reach the intracellular space.

Double glomus tumors originating in the submandibular and parotid regions.

Glomus tumors are rare neoplasms that originate from the glomus bodies, an arteriovenous anastomosis with a specialized vascular structure. The most common site for these tumors is the subungal region of the fingers. Occasionally, glomus tumors are found in the middle ear, trachea, nasal cavities, stomach, and lungs. The occurrence in the parotid regions is very rare. While multiple glomus tumors in the whole body are thought to represent only 10% of all cases, instances of multiple tumors in the neck have not yet been reported in the literature. We report a case of double glomus tumors in the submandibular and parotid regions.

Assessment of open versus laparoscopy-assisted gastrectomy in lymph node-positive early gastric cancer: a retrospective cohort analysis.

BACKGROUND: Laparoscopy-assisted gastrectomy (LAG) is still limited for early gastric cancer (EGC) with low possibility of lymph node (LN) metastasis, due to the concern for incomplete LN dissection and controversial long-term outcomes. We assessed oncological outcomes of laparoscopy-assisted versus open gastrectomy (OG) for patients with LN positive EGC. METHODS: Between 2003 and 2007, 204 patients underwent surgery for LN positive EGC. We evaluated adequacy of LN dissection and early and long-term outcomes after OG (n = 162) and LAG (n = 42). RESULTS: Operative time was longer but hospital stay was shorter for LAG than OG. Postoperative complications occurred in 14 patients (8.6%) after OG and 1 patient (2.4%) after LAG (P = 0.316). Mean number of retrieved LNs and number of retrieved and metastatic LNs for each station did not differ between the two groups. During median 35 months of follow-up, 14 patients (8.6%) developed recurrence after OG, compared with 4 patients (9.5%) after LAG (P = 0.769). Overall 5-year disease-free survival was 89.9% and 89.7% after OG and LAG. Status of LN metastasis was the only independent prognostic factor for disease-free survival. CONCLUSIONS: LAG is an oncologically safe procedure even for LN positive EGC. Adequate LN dissection and comparable long-term outcomes to OG can be achieved by LAG.

Balloon catheter assisted excision of benign cervical cyst.

OBJECTIVES: Benign cystic masses that develop in the neck are easily excised with a wide skin incision. However, if the cyst is perforated and the contour of the cyst is not maintained during surgery, complete excision may be difficult. We developed a surgical method that can maintain the contour of a cyst and decrease the wound size with a balloon catheter. METHODS: Two cases with a branchial cleft cyst (BCC) were removed using this method without recurrence. The surgical technique includes a skin incision above the cyst, and fine dissection to expose the cyst wall. Then a small tap incision was created on the cyst using a #11 blade and a #14 Fr. Foley catheter was inserted through the incision site and the balloon was inflated up to 4 cm(3) using normal saline. The fluid inner content was carefully removed keeping the intracystic balloon inflated. By pulling and releasing the catheter, the cyst was dissected carefully and excised. RESULTS: Complete excision of the cyst was performed with the balloon catheter successfully. The final pathological diagnosis was BCC in the two cases. CONCLUSIONS: The balloon-assisted technique was easy to perform and could be used for the excision of a benign cystic neck mass.

Robotic gastrectomy for gastric cancer: surgical techniques and clinical merits.

BACKGROUND: Robotic gastrectomy in the setting of gastric cancer is reported by some investigators. However, no study has compared robotic surgery with open or laparoscopic surgery for patients with gastric cancer. This study aimed to determine the clinical benefits of robotic gastrectomy over open and laparoscopic gastrectomy for the treatment of gastric cancer. METHODS: After the introduction of the da Vinci surgical system in November 2007 at the authors' hospital, 18 robotic gastrectomies were performed from 31 December 2007 to 30 June 2008. The prospective data from gastric cancer patients who underwent gastrectomies (16 robotic, 11 laparoscopic, and 12 open) during the same period were retrospectively analyzed. RESULTS: Sex, age, comorbidity, extent of lymphadenectomy, pT stage, lymph node metastasis, and number of lymph nodes retrieved were similar among the three groups. The estimated blood loss was significantly less in the robotic gastrectomy group than in the open group (p = 0.0312), and the postoperative hospital stay in the robotic group was significantly shorter than in the open and laparoscopic gastrectomy groups (p < 0.001). Postoperative morbidity and time to first flatus were similar in the three groups. There was no open or laparoscopic conversion in the robotic group. No postoperative mortality occurred in any group. CONCLUSION: Robotic gastrectomy for the treatment of gastric cancer is a feasible and safe procedure in the hands of experienced laparoscopic surgeons. Robotic gastrectomy offers better short-term surgical outcomes than the open and laparoscopic methods. Furthermore, this procedure may be a preferable alternative for the treatment of gastric cancer.

A novel modification of double stapling technique in Billroth I anastomosis.

Although the double-stapling technique has been used as a safe procedure in gastroduodenostomy, creating anastomosis with a circular stapler on the linear stapler line can be vulnerable for anastomotic leakage and ischemia. Therefore, we tried to modify the double-stapling technique to avoid stapling on the staple line.