Decoding cellular mechanism of recombinant adeno-associated virus (rAAV) and engineering host-cell factories toward intensified viral vector manufacturing.

Recombinant adeno-associated virus (rAAV) is one of the prominent gene delivery vehicles that has opened promising opportunities for novel gene therapeutic approaches. However, the current major viral vector production platform, triple transfection in mammalian cells, may not meet the increasing demand. Thus, it is highly required to understand production bottlenecks from the host cell perspective and engineer the cells to be more favorable and tolerant to viral vector production, thereby effectively enhancing rAAV manufacturing. In this review, we provided a comprehensive exploration of the intricate cellular process involved in rAAV production, encompassing various stages such as plasmid entry to the cytoplasm, plasmid trafficking and nuclear delivery, rAAV structural/non-structural protein expression, viral capsid assembly, genome replication, genome packaging, and rAAV release/secretion. The knowledge in the fundamental biology of host cells supporting viral replication as manufacturing factories or exhibiting defending behaviors against viral production is summarized for each stage. The control strategies from the perspectives of host cell and materials (e.g., AAV plasmids) are proposed as our insights based on the characterization of molecular features and our existing knowledge of the AAV viral life cycle, rAAV and other viral vector production in the Human embryonic kidney (HEK) cells.

Feasibility of a Novel In-situ Local Tumor Ablation and Recycling Machine Based on Radiofrequency Dielectric Heating: In-depth Review on Research Background and Preliminary Report of an Experimental Study.

Background: In bone sarcomas, chemotherapy has improved the prognosis with advances in diagnostic and surgical technologies, which has led to attempts to save limbs. As early detection and multidisciplinary treatment have improved the survival rate, curative surgery is considered for selected patients with metastatic bone carcinomas. Limb salvage procedures may vary in relation to the reconstruction method, which is accompanied by different complications. To overcome them, we devised a novel concept, in-situ local tumor ablation and recycling machine based on radiofrequency (RF)-induced heating and intended experiments to demonstrate its feasibility. Methods: The fresh femurs of 6-month-old pigs were used after removing the epiphyses; the distal parts were placed in a heating chamber. Fiber-optic temperature sensors were inserted in the metaphysis, meta-diaphysis, and diaphysis. Temperatures were measured six times each during heating at 27.12 MHz at various powers. Additionally, the compressive and bending stiffnesses were measured six times each for the unprocessed, RF-treated, and pasteurized bones, and the results were compared. Results: Under 200 W power output, the temperatures at all measurement sites reached 70 ℃ or higher in 6 minutes, and the temperatures were maintained. The median compressive stiffness of RF-heated bones was 79.2% higher than that of pasteurized bones, but the difference was statistically insignificant. The median bending stiffness of RF-heated bones was approximately 66.3% of that of unprocessed bones, which was 20% higher than that of pasteurized bones. Conclusions: The feasibility to rapidly attain and maintain temperatures for tumor ablation is shown, which favorably preserves bone stiffness through the in-situ local tumor ablation and recycling based on RF heating. The problem of nonuniform temperature distribution might be solved by an optimal design determined from simulation research and additional experiments.

Shewanella goraebulensis sp. nov., isolated from sea water.

A novel Gram-stain-negative, facultatively anaerobic and rod-shaped bacterial strain, designated as DAU312T, was isolated from the sea water of the eastern coast of the Republic of Korea. Optimal growth was observed at 25 °C, pH 7.0-8.0 and with NaCl concentrations of 2.0 % (w/v). Catalase and oxidase activities were detected. On the basis of 16S rRNA gene sequences, strain DAU312T showed the highest similarity (99.2 %) to the type strain Shewanella electrodiphila MAR441T. The complete genome sequence of strain DAU312T contains 4 893 483 bp and 40.5 mol% G+C. Phylogenetic analyses based on 16S rRNA gene sequences and the up-to-date bacterial core genes showed that strain DAU312T, S. electrodiphila MAR441T and S. olleyana were all part of the same monophyletic clade. Their average nucleotide identity, digital DNA-DNA hybridization and two-way average amino acid identity values with each other and type strains of close Shewanella species were 83.4-77.5 %, 27.3-22.0 % and 89.8-81.2 %, respectively. The major cellular fatty acids (>10 %) were iso-C15 : 0, summed feature 3 (C16 : 1 ω7с and/or C16 : 1 ω6с) and C16 : 0. Phosphatidylethanolamine and phosphatidylglycerol were the main polar lipids. The respiratory quinones were Q-7, Q-8, MK-7 and MMK-7. Based on these polyphasic taxonomic findings, the name Shewanella goraebulensis sp. nov. is suggested for strain DAU312T, which is considered to represent a novel species of the genus Shewanella. The type strain is DAU312T (=KCTC 72427 T=JCM 35744T=KCCM 43478T).

Current Treatment Concepts for Extra-Abdominal Desmoid-Type Fibromatosis: A Narrative Review.

Extra-abdominal desmoid-type fibromatosis (EADTF) is a rare neoplastic condition of monoclonal fibroblastic proliferation characterized by local aggressiveness with a distinct tendency to recur. Although EADTF is a benign disease entity, these tumors have a tendency to infiltrate surrounding normal tissues, making it difficult to completely eliminate them without adjacent healthy tissue injury. Surgical excision of these locally aggressive tumors without clear resection margins often leads to local recurrence. The aim of this thorough review was to assess the current treatment concepts for these rare tumors. A comprehensive search of articles published in the Cochrane Library, MEDLINE (PubMed), and EMBASE databases between January 2008 and February 2023 was conducted. Surgical intervention is no longer the first-line approach for most cases; instead, strategies like active surveillance or systemic therapies are used as initial treatment options. With the exception of EADTFs situated near vital structures, a minimum of 6-12 months of active surveillance is currently advocated for, during which some disease progression may be considered acceptable. Non-surgical interventions such as radiation or cryoablation may be employed in certain patients to achieve local control. The currently preferred systemic treatment options include tyrosine kinase inhibitors, low-dose chemotherapy, and gamma-secretase inhibitors, while hormone therapy is not advised. Nonsteroidal anti-inflammatory drugs are utilized primarily for pain management.

Measuring urban quality and change through the detection of physical attributes of decay.

The quality of the urban environment is crucial for societal well-being. Yet, measuring and tracking the quality of urban environment, their evolution, and spatial disparities is difficult due to the amount of on-the-ground data needed to capture these patterns. The growing availability of street view images presents new prospects in identifying urban features. However, the reliability and consistency of these methods across different locations and time remains largely unexplored. We aim to develop a comprehensive index of urban quality and change at the street segment level using Google Street View (GSV) imagery. We focus on eight object classes that indicate urban decay or contribute to an unsightly urban space, such as potholes, graffiti, garbage, tents, barred or broken windows, discolored or dilapidated façades, weeds, and utility markings. We train an object detection model on a dataset of images from different cities and assess the performance of these urban indices. We evaluate the effectiveness of this method in various urban contexts over time and discuss its potential for urban planning and public policy. We demonstrate the use of these indices in three applications: the Tenderloin in San Francisco, the Doctores and Historic Center neighborhoods in Mexico City, and South Bend, Indiana.

Marine plant-based biorefinery for sustainable 2,5-furandicarboxylic acid production: A review.

Marine plants, including macroalgae and seagrass, show promise as biorenewable feedstocks for sustainable chemical manufacturing. This study explores their potential in producing 2,5-furandicarboxylic acid (FDCA), a versatile platform chemical for commodity polymers. FDCA-based polyethylene 2,5-furandicarboxylate offers a sustainable alternative to petroleum-derived polyethylene terephthalate, commonly used in plastic bottles. Our research pioneers the concept of a marine plant-based FDCA biorefinery, introducing innovative approaches for sustainability and cost-effectiveness. This review outlines the use of ionic liquid-based solvents (ILS) and deep eutectic solvent (DES) systems in FDCA production. Additionally, we propose biomodification strategies involving target enzyme-encoding genes to enhance the depolymerization of non-structural storage glucans in marine plants. Our findings pave the way for eco-friendly biorefineries and biorenewable plastics.

Development and Validation of a Convolutional Neural Network Model to Predict a Pathologic Fracture in the Proximal Femur Using Abdomen and Pelvis CT Images of Patients With Advanced Cancer.

BACKGROUND: Improvement in survival in patients with advanced cancer is accompanied by an increased probability of bone metastasis and related pathologic fractures (especially in the proximal femur). The few systems proposed and used to diagnose impending fractures owing to metastasis and to ultimately prevent future fractures have practical limitations; thus, novel screening tools are essential. A CT scan of the abdomen and pelvis is a standard modality for staging and follow-up in patients with cancer, and radiologic assessments of the proximal femur are possible with CT-based digitally reconstructed radiographs. Deep-learning models, such as convolutional neural networks (CNNs), may be able to predict pathologic fractures from digitally reconstructed radiographs, but to our knowledge, they have not been tested for this application. QUESTIONS/PURPOSES: (1) How accurate is a CNN model for predicting a pathologic fracture in a proximal femur with metastasis using digitally reconstructed radiographs of the abdomen and pelvis CT images in patients with advanced cancer? (2) Do CNN models perform better than clinicians with varying backgrounds and experience levels in predicting a pathologic fracture on abdomen and pelvis CT images without any knowledge of the patients' histories, except for metastasis in the proximal femur? METHODS: A total of 392 patients received radiation treatment of the proximal femur at three hospitals from January 2011 to December 2021. The patients had 2945 CT scans of the abdomen and pelvis for systemic evaluation and follow-up in relation to their primary cancer. In 33% of the CT scans (974), it was impossible to identify whether a pathologic fracture developed within 3 months after each CT image was acquired, and these were excluded. Finally, 1971 cases with a mean age of 59 ± 12 years were included in this study. Pathologic fractures developed within 3 months after CT in 3% (60 of 1971) of cases. A total of 47% (936 of 1971) were women. Sixty cases had an established pathologic fracture within 3 months after each CT scan, and another group of 1911 cases had no established pathologic fracture within 3 months after CT scan. The mean age of the cases in the former and latter groups was 64 ± 11 years and 59 ± 12 years, respectively, and 32% (19 of 60) and 53% (1016 of 1911) of cases, respectively, were female. Digitally reconstructed radiographs were generated with perspective projections of three-dimensional CT volumes onto two-dimensional planes. Then, 1557 images from one hospital were used for a training set. To verify that the deep-learning models could consistently operate even in hospitals with a different medical environment, 414 images from other hospitals were used for external validation. The number of images in the groups with and without a pathologic fracture within 3 months after each CT scan increased from 1911 to 22,932 and from 60 to 720, respectively, using data augmentation methods that are known to be an effective way to boost the performance of deep-learning models. Three CNNs (VGG16, ResNet50, and DenseNet121) were fine-tuned using digitally reconstructed radiographs. For performance measures, the area under the receiver operating characteristic curve, accuracy, sensitivity, specificity, precision, and F1 score were determined. The area under the receiver operating characteristic curve was used to evaluate three CNN models mainly, and the optimal accuracy, sensitivity, and specificity were calculated using the Youden J statistic. Accuracy refers to the proportion of fractures in the groups with and without a pathologic fracture within 3 months after each CT scan that were accurately predicted by the CNN model. Sensitivity and specificity represent the proportion of accurately predicted fractures among those with and without a pathologic fracture within 3 months after each CT scan, respectively. Precision is a measure of how few false-positives the model produces. The F1 score is a harmonic mean of sensitivity and precision, which have a tradeoff relationship. Gradient-weighted class activation mapping images were created to check whether the CNN model correctly focused on potential pathologic fracture regions. The CNN model with the best performance was compared with the performance of clinicians. RESULTS: DenseNet121 showed the best performance in identifying pathologic fractures; the area under the receiver operating characteristic curve for DenseNet121 was larger than those for VGG16 (0.77 ± 0.07 [95% CI 0.75 to 0.79] versus 0.71 ± 0.08 [95% CI 0.69 to 0.73]; p = 0.001) and ResNet50 (0.77 ± 0.07 [95% CI 0.75 to 0.79] versus 0.72 ± 0.09 [95% CI 0.69 to 0.74]; p = 0.001). Specifically, DenseNet121 scored the highest in sensitivity (0.22 ± 0.07 [95% CI 0.20 to 0.24]), precision (0.72 ± 0.19 [95% CI 0.67 to 0.77]), and F1 score (0.34 ± 0.10 [95% CI 0.31 to 0.37]), and it focused accurately on the region with the expected pathologic fracture. Further, DenseNet121 was less likely than clinicians to mispredict cases in which there was no pathologic fracture than cases in which there was a fracture; the performance of DenseNet121 was better than clinician performance in terms of specificity (0.98 ± 0.01 [95% CI 0.98 to 0.99] versus 0.86 ± 0.09 [95% CI 0.81 to 0.91]; p = 0.01), precision (0.72 ± 0.19 [95% CI 0.67 to 0.77] versus 0.11 ± 0.10 [95% CI 0.05 to 0.17]; p = 0.0001), and F1 score (0.34 ± 0.10 [95% CI 0.31 to 0.37] versus 0.17 ± 0.15 [95% CI 0.08 to 0.26]; p = 0.0001). CONCLUSION: CNN models may be able to accurately predict impending pathologic fractures from digitally reconstructed radiographs of the abdomen and pelvis CT images that clinicians may not anticipate; this can assist medical, radiation, and orthopaedic oncologists clinically. To achieve better performance, ensemble-learning models using knowledge of the patients' histories should be developed and validated. The code for our model is publicly available online at https://github.com/taehoonko/CNN_path_fx_prediction . LEVEL OF EVIDENCE: Level III, diagnostic study.

Design space determination to optimize DNA complexation and full capsid formation in transient rAAV manufacturing.

Recombinant adeno-associated virus (rAAV) vectors are a promising platform for in vivo gene therapies. However, cost-effective, well-characterized processes necessary to manufacture rAAV therapeutics are challenging to develop without an understanding of how process parameters (PPs) affect rAAV product quality attributes (PQAs). In this work, a central composite orthogonal experimental design was employed to examine the influence of four PPs for transient transfection complex formation (polyethylenimine:DNA [PEI:DNA] ratio, total DNA/cell, cocktail volume, and incubation time) on three rAAV PQAs related to capsid content (vector genome titer, vector genome:capsid particle ratio, and two-dimensional vector genome titer ratio). A regression model was established for each PQA using partial least squares, and a design space (DS) was defined in which Monte Carlo simulations predicted < 1% probability of failure (POF) to meet predetermined PQA specifications. Of the three PQAs, viral genome titer was most strongly correlated with changes in complexation PPs. The DS and acceptable PP ranges were largest when incubation time and cocktail volume were kept at mid-high setpoints, and PEI:DNA ratio and total DNA/cell were at low-mid setpoints. Verification experiments confirmed model predictive capability, and this work establishes a framework for studying other rAAV PPs and their relationship to PQAs.

Critical challenges and advances in recombinant adeno-associated virus (rAAV) biomanufacturing.

Gene therapy is a promising therapeutic approach for genetic and acquired diseases nowadays. Among DNA delivery vectors, recombinant adeno-associated virus (rAAV) is one of the most effective and safest vectors used in commercial drugs and clinical trials. However, the current yield of rAAV biomanufacturing lags behind the necessary dosages for clinical and commercial use, which embodies a concentrated reflection of low productivity of rAAV from host cells, difficult scalability of the rAAV-producing bioprocess, and high levels of impurities materialized during production. Those issues directly impact the price of gene therapy medicine in the market, limiting most patients' access to gene therapy. In this context, the current practices and several critical challenges associated with rAAV gene therapy bioprocesses are reviewed, followed by a discussion of recent advances in rAAV-mediated gene therapy and other therapeutic biological fields that could improve biomanufacturing if these advances are integrated effectively into the current systems. This review aims to provide the current state-of-the-art technology and perspectives to enhance the productivity of rAAV while reducing impurities during production of rAAV.

Transcriptomic features reveal molecular signatures associated with recombinant adeno-associated virus production in HEK293 cells.

The development of gene therapies based on recombinant adeno-associated viruses (rAAVs) has grown exponentially, so the current rAAV manufacturing platform needs to be more efficient to satisfy rising demands. Viral production exerts great demand on cellular substrates, energy, and machinery; therefore, viral production relies heavily on the physiology of the host cell. Transcriptomics, as a mechanism-driven tool, was applied to identify significantly regulated pathways and to study cellular features of the host cell for supporting rAAV production. This study investigated the transcriptomic features of two cell lines cultured in their respective media by comparing viral-producing cultures with non-producing cultures over time in parental human embryonic kidney cells (HEK293). The results demonstrate that the innate immune response signaling pathways of host cells (e.g., RIG-I-like receptor signaling pathway, Toll-like receptor signaling pathway, cytosolic DNA sensing pathway, JAK-STAT signaling pathway) were significantly enriched and upregulated. This was accompanied by the host cellular stress responses, including endoplasmic reticulum stress, autophagy, and apoptosis in viral production. In contrast, fatty acid metabolism and neutral amino acid transport were downregulated in the late phase of viral production. Our transcriptomics analysis reveals the cell-line independent signatures for rAAV production and serves as a significant reference for further studies targeting the productivity improvement in the future.

Minimally Invasive Carpal Tunnel Release Using a Hook Knife through a Small Transverse Carpal Incision: Technique and Outcome.

Background: Compared to the classic open technique, limited exposure techniques have the advantages of less scar pain, more grip and pinch strength, and earlier return to daily life. We evaluated the effectiveness and safety of our novel method of minimally invasive carpal tunnel release using a hook knife through a small transverse carpal incision. Methods: This study included 111 carpal tunnel decompressions in 78 patients who underwent carpal tunnel release from January 2017 to December 2018. We performed carpal tunnel release using a hook knife through a small transverse incision just proximal to the wrist crease under local infiltration of lidocaine with tourniquet inflation in the upper arm. All patients were tolerable during the procedure and discharged following the procedure on the same day. Results: At an average of 29.4 months of follow-up (range, 12-51 months), all but one patient (99%) revealed complete or near complete symptomatic recovery. The average of the symptom severity score on the Boston questionnaire was 1.31 ± 0.30 and the average of the functional status score was 1.19 ± 0.26. The final mean Quick Disabilities of the Arm, Shoulder, and Hand questionnaire (QuickDASH) score was 8.66 (range, 2-39). There was no superficial palmar arch injury or nerve injury of the palmar cutaneous branch, recurrent motor branch, or median nerve itself as a complication of the procedure. No patient showed any wound infection or wound dehiscence. Conclusions: Our carpal tunnel release using a hook knife through a small transverse carpal incision performed by an experienced surgeon is a safe and reliable method that is expected to have the benefits of simplicity and minimal invasiveness.

Retrodural space of Okada in the posterior ligamentous complex region: clinical and anatomical findings relevant to lumbar interlaminar epidural injection.

BACKGROUND: The retrodural space of Okada is a potential space posterior to the ligamentum flavum that allows communication with the bilateral facet joints. However, the actual anatomy of this space has not been clearly visualized to date. We sought to investigate the characteristics of patients showing contrast spreading to the facet joint space during epidural injection and to clarify the anatomical structures of the retrodural space and adjacent ligamentous tissues in cadaveric specimens. METHODS: Fluoroscopic images of patients who underwent fluoroscopy-guided lumbar interlaminar epidural injection were assessed for contrast flow to the facet joints. Patient demographics, preprocedural imaging study findings, and epidural approaches were analyzed. The anatomical study included the sectional dissection, micro-CT imaging, and histological evaluation of lumbar spine specimens from 16 embalmed cadavers. RESULTS: Fluoroscopic images of 605 epidural injections were analyzed. Among them, 36 with inadvertent spread into the facet joints (5.9%) were identified. Multivariate analysis revealed that facet joint pathologies were significantly associated with inadvertent spread into the facet joints (OR 4.382; 95% CI 1.160 to 16.558; p=0.029). Micro-CT and histological findings consistently showed a retrodural space between the ligamentum flavum and interspinous ligament. Various anatomical communication routes in the posterior ligamentous complex leading to this space were observed in specimens with degenerative and pathological changes. CONCLUSION: Degenerative and pathological facet joint changes were associated with a higher incidence of spread into the retrodural space during epidural injection. Our findings confirm anatomical evidence for a false loss of resistance before the needle enters the epidural space.

Strategic biomodification for raw plant-based pretreatment biorefining toward sustainable chemistry.

Plant-based pretreatment biorefining is the initial triggering process in biomass-conversion to bio-based chemical products. In view of chemical sustainability, the raw plant-based pretreatment biorefining process is more favorable than the fossil-based one. Its direct use contributes to reducing CO2 emissions and the production cost of the target products by eliminating costly steps, such as the separation and purification of intermediates. Three types of feedstock plant resources have been utilized as raw plant feedstock sources, such as: lignocellulosic, starchy, and inulin-rich feedstock plants. These plant sources can be directly used for bio-based chemical products. To enhance the efficiency of their pretreatment biorefining process, well-designed biomodification schemes are discussed in this review to afford important information on useful biomodification approaches. For lignocellulosic feedstock plants, the enzymes and regulatory elements involved in lignin reduction are discussed using: COMT, GAUT4, CSE, PvMYB4 repressor, etc. For inulin-rich feedstock plants, 1-SST, 1-FFT, 1-FEH, and endoinulinase are illustrated in relation with the reduction of chain length of inulin polymer. For starchy feedstock plants, their biomodification is targeted to enhancing the depolymerization efficiency of starch to glucose monomer units. For this biomodification target, six candidates are discussed. These are SBE I, SBE IIa, SBE IIb, GBSS I, PTSTI, GWD 1, and PTSTI. The biomodification strategies discussed here promise to be conducive to enhancing the efficiency of the plant-based pretreatment biorefining process.

Prevalence of Vitamin D Deficiency in Children with Fractures: Before and during the COVID-19 Outbreak.

Background: Although it is generally agreed that vitamin D is important for bone health, the role of vitamin D in preventing fractures in children and adolescents remains unclear. Therefore, this study aimed to investigate the prevalence of vitamin D deficiency and insufficiency in healthy Korean children with fractures. Our secondary aim was to compare serum vitamin D levels before and during the coronavirus disease 2019 (COVID-19) outbreak. Methods: We evaluated 334 patients with fractures who were surgically treated at our institution between 2018 and 2019 before the onset of COVID-19 (group I). In addition, we collected data on the serum 25(OH)D levels of 210 patients who visited our pediatric department for evaluation of short stature (group II) and the serum 25(OH)D levels of the patients with fractures during the COVID-19 pandemic period (group III). A serum 25(OH)D level of <20 ng/mL was considered deficient, between 20 and 32 ng/mL was insufficient, and ≥32 ng/mL was considered sufficient. Results: The mean age was 8.1 ± 3.5 years in group I, 8.2 ± 3.7 years in group II, and 8.6 ± 3.5 years in group III. The prevalence of vitamin D deficiency was 53.0% in group I and 32.9% in group II. The mean serum 25(OH)D level was lower in group I than in group II (20.0 ± 7.3 ng/ml vs. 23.2 ± 6.9 ng/ml, p < 0.001). The mean serum 25(OH)D level of younger patients (<10 years) in group III was lower than that of the younger patients in the prepandemic period (21.4 ± 7.2 ng/mL vs. 19.2 ± 6.8 ng/mL, p=0.037). Conclusions: We observed a high prevalence of vitamin D deficiency/insufficiency in children with fractures who required surgical treatment. During the COVID-19 pandemic, the serum vitamin D levels became even lower, especially in younger children.

Secondary Malignancy in Giant Cell Tumor: A Single-Center Study.

Giant cell tumor of bone (GCTB) undergoes a sarcomatous transformation. Secondary malignancy in giant cell tumor (MGCT) is associated with radiotherapy and has a dismal prognosis. We reviewed medical records to investigate the clinicopathological characteristics and prognosis of MGCT patients. The enrollment criterion was high-grade spindle-cell sarcoma, which developed at the site of prior GCTB treatment. Twelve patients were analyzed: six females and six males. The median age was 42.5 years. Benign recurrence occurred in five GCTB patients not treated with radiotherapy. No pulmonary implants were observed. The median latency to the malignant transformation was 63 months. Nine patients were AJCC stage IIB, and three were stage IVA. The median follow-up period after malignant transformation was 62.5 months. Five patients developed local recurrence, and six had distant metastasis. Five-year overall recurrence and metastasis-free survival rates were 61.9%, 66.7%, and 58.3%, respectively. Initial metastasis was a predictive factor for overall survival. Benign local recurrence of GCTB was also a negative factor for metastasis-free survival of MGCT patients. Differences in overall survival according to benign recurrence also showed a tendency toward significance. In our series, secondary MGCT did not occur after radiotherapy. The prognosis was better than previous findings. Benign recurrence of GCTB could reflect the prognosis of MGCT.

Sarcomas in Teachers Using Three-Dimensional Printers: A Report of Three Patients and Literature Review.

Background: While low-cost, small-scale, desktop three-dimensional (3D) printers are gaining popularity in the education sector, some studies have reported harmful emissions of particles and volatile organic compounds during the fused deposition modeling (FDM) process, posing a potential health risk. Sarcomas are rare tumors, constituting a group of diverse rare malignant tumors. While some genetic and environmental factors contribute to the development of sarcomas, most cases are idiopathic and sporadic. Methods: We secured the medical records and statements about work environment from teachers diagnosed with sarcomas after frequent use of 3D printers in high schools, reviewed the cases, and described them in narrative format. Furthermore, popularization of FDM 3D printers, worrisome emissions released during the printing process, and related precautions and countermeasures were discussed through literature review. Results: Exceptionally, the cases of sarcomas, such as Ewing's sarcoma, malignant peripheral nerve sheath tumor, and well-differentiated liposarcoma, arose in a common specific condition. All the teachers regularly operated 3D printers in poorly ventilated spaces for at least 2 years. They had no past or family history of relevant diseases. Conclusions: We first reported three cases of sarcoma in teachers who used 3D printers in poorly ventilated conditions. Although a relationship between the use of 3D printers and the development of sarcomas has not been determined yet, it is important to come up with measures to protect teachers and students using 3D printers from the potential hazard.

Effect of Preoperative Oral Carbohydrates on Insulin Resistance in Older Adults Who Underwent Total Hip or Knee Arthroplasty: A Prospective Randomized Trial.

INTRODUCTION: Preoperative carbohydrate drinks are recommended to reduce fasting time before surgery. Older adults are at risk of pulmonary aspiration and hyperglycemia after consuming carbohydrate drinks because of increased insulin resistance and delayed gastric emptying. We investigated the effects of oral carbohydrate drinks on perioperative insulin resistance, metabolic responses, and gastric volume in older adults. METHODS: Fifty-six patients (aged more than 65 years) were randomly assigned to the control or carbohydrate (CHO) group. The CHO group received 400 mL of a carbohydrate drink 2 to 3 hours before anesthesia. The control group was allowed clear fluid intake 2 hours before anesthesia. Blood glucose and insulin levels were measured before intake of the carbohydrate drink and 1 hour postoperatively. Gastric volume was measured before spinal anesthesia. Insulin resistance was calculated using the homeostasis model assessment for insulin resistance. RESULTS: Homeostasis model assessment for insulin resistance was not different between the control and CHO groups preoperatively (2.5 versus 3.3, P = 0.156) or postoperatively (2.6 versus 2.4, P = 0.817). Preoperative gastric volume was comparable between the control and CHO groups (35.5 versus 30.8 mL, P = 0.696). DISCUSSION: Preoperative oral consumption of carbohydrates did not affect insulin resistance or gastric volume in older adults undergoing total knee or hip arthroplasty. Preoperative carbohydrate loading is safe in older adults undergoing total knee or hip arthroplasty. DATA AVAILABILITY: The data that support the findings of this study are available from the corresponding author on reasonable request. CLINICAL TRIAL REGISTRATION: ClinicalTrials.gov (No. NCT04206189).

Optimal location for continuous catheter analgesia among the femoral triangle, proximal, or distal adductor canal after total knee arthroplasty: a randomized double-blind controlled trial.

BACKGROUND: Pain management after total knee arthroplasty is essential to improve early mobilization, rehabilitation, and recovery. Continuous adductor canal (AC) block provides postoperative analgesia while preserving quadriceps strength. However, there have been inconsistencies regarding the optimal location for continuous catheter block. We compared continuous femoral triangle, proximal AC, and distal AC blocks for postoperative analgesia after total knee arthroplasty. METHODS: Patients undergoing unilateral total knee arthroplasty were randomly assigned to three groups: femoral triangle, proximal AC, or distal AC. The surgeon performed periarticular local anesthetic infiltration. After surgery, an ultrasound-guided perineural catheter insertion procedure was performed. The primary endpoint was pain scores at rest in the morning on the first postoperative day. Secondary endpoints included pain scores at rest and during activity at other time points, quadriceps strength, and opioid consumption. RESULTS: Ninety-five patients, 32 in the femoral triangle group, 31 in the proximal AC group, and 32 in the distal AC group, completed the study. Analysis of the primary outcome showed no significant difference in pain scores among groups. Secondary outcomes showed significantly lower pain scores at rest and during activity in the distal AC group than in the femoral triangle and proximal AC groups in the morning of the second postoperative day. Quadriceps strength and opioid consumption did not differ among groups. CONCLUSIONS: Continuous femoral triangle, proximal AC, and distal AC blocks in the setting of periarticular local anesthetic infiltration provide comparable postoperative analgesia after total knee arthroplasty. TRIAL REGISTRATION NUMBER: NCT04206150.

Effects of Anesthetic Techniques on the Risk of Postoperative Complications Following Lower Extremity Amputation in Diabetes Patients with Coagulation Abnormalities: A Retrospective Cohort Study Using Propensity Score Analysis.

Diabetic foot amputation is associated with high morbidity and mortality rates. To prevent cardiovascular complications along with vasculopathy in the course of diabetes mellitus, a high number of patients receive anticoagulant therapy. However, anticoagulants are contraindicated in neuraxial anesthesia limiting available anesthetic modalities. Therefore, in this retrospective study, we aimed to compare between general anesthesia and peripheral nerve block (PNB) with respect to postoperative complications following lower extremity amputation (LEA) in patients with coagulation abnormalities. In total, 320 adult patients who underwent LEA for diabetic foot were divided into two groups according to the anesthetic type (general anesthesia vs. PNB). The inverse probability of treatment weighting was performed to balance the baseline patient characteristics and surgical risk between the two groups. The adjusted analysis showed that compared with the general anesthesia group, the PNB group had lower risks of pneumonia (odds ratio: 0.091, 95% confidence interval [CI]: 0.010-0.850, p = 0.0355), acute kidney injury (odds ratio: 0.078, 95% CI: 0.007-0.871, p = 0.0382), and total major complications (odds ratio: 0.603, 95% CI: 0.400-0.910, p = 0.0161). Additionally, general anesthesia was associated with a higher amount of intraoperative crystalloid administration and a requirement for more frequent vasopressors. In conclusion, PNB appears to be protective against complications following LEA in diabetes patients with coagulopathy.

Clostridium acetobutylicum atpG-Knockdown Mutants Increase Extracellular pH in Batch Cultures.

ATPase, a key enzyme involved in energy metabolism, has not yet been well studied in Clostridium acetobutylicum. Here, we knocked down the atpG gene encoding the ATPase gamma subunit in C. acetobutylicum ATCC 824 using a mobile group II intron system and analyzed the physiological characteristics of the atpG gene knockdown mutant, 824-2866KD. Properties investigated included cell growth, glucose consumption, production of major metabolites, and extracellular pH. Interestingly, in 2-L batch fermentations, 824-2866KD showed no significant difference in metabolite biosynthesis or cell growth compared with the parent ATCC 824. However, the pH value in 824-2866KD cultures at the late stage of the solventogenic phase was abnormally high (pH 6.12), compared with that obtained routinely in the culture of ATCC 824 (pH 5.74). This phenomenon was also observed in batch cultures of another C. acetobutylicum, BEKW-2866KD, an atpG-knockdown and pta-buk double-knockout mutant. The findings reported in this study suggested that ATPase is relatively minor than acid-forming pathway in ATP metabolism in C. acetobutylicum.