Reconfiguring the Escherichia coli Electron Transport Chain to Enhance trans-2-Decenoic Acid Production.

trans-2-Decenoic acid is a pivotal α,ß-medium-chain unsaturated fatty acid that serves as an essential intermediary in the synthesis of 10-hydroxy-2-decenoic acid and various pharmaceutical compounds. Biosynthesis yield of trans-2-decenoic acid by decanoic acid has significantly improved in recent years; however, the oxidative stress of Escherichia coli at high fatty acid concentrations restricts the conversion rate. Here, we introduced a combination of rational design and metabolic rewiring of the E. coli electron transport chain (ETC) to improve trans-2-decenoic acid production. Overexpressing ubiquinone (UbQ) biosynthesis genes enhanced the expression of ETC complex III: UbQ to reduce reactive oxygen species (ROS) accumulation. Furthermore, applying rotenone to inhibit ETC complex I improved the electron transfer efficiency of complex II. The integration of Vitamin B5 and B2 into the fermentation process increased the activities of fatty acyl-CoA synthetase (MaMACS) and fatty acyl-CoA dehydrogenase (PpfadE). Finally, the constructed E. coli BL21(DE3)(ΔfadBJR/pCDFDuet-1-PpfadE-MaMACS/pRSFDuet-1-sumo-CtydiI-ubiI) strain exhibited a 51.50% decrease in ROS and a 93.33% enhancement in trans-2-decenoic acid yield, reaching 1.45 g/L after 66 h, which is the highest yield reported for flask fermentation. This study reports the feasibility of rewiring the ETC regulation and energy metabolism to improve α,ß-UCA biosynthesis efficiency.

Effect of lower-limb isokinetic muscle strengthening on knee function and joint contact force in knee osteoarthritis patients awaiting total knee arthroplasty: study protocol for a randomized controlled trial.

BACKGROUND: Knee osteoarthritis (OA) is a common and serious joint disease and patients mainly suffer from knee pain and dysfunction, significantly impacting their quality of life and daily activities. Non-pharmacological treatments and total knee arthroplasty (TKA) are the two major treatments for knee OA. TKA is the primary treatment for severe knee OA patients, however, the postoperative rehabilitation results are not entirely satisfactory. To enhance postoperative rehabilitation outcomes, preoperative lower-limb muscle strengthening has aroused attention and demonstrated significant results on functional recovery after TKA. Specifically, isokinetic muscle strengthening (IMS) has been highly anticipated as an efficient training method. Nevertheless, its effects in perioperative and long-term periods of TKA require further investigation. This study aims to evaluate the impact of preoperative lower-limb IMS during the 4 weeks before TKA on the perioperative and postoperative outcomes, with particular attention on knee function, pain, and inflammatory response. METHODS: In this study, 104 patients awaiting TKA will be randomized to 2 groups: the IMS and health education groups. All patients will be evaluated at baseline, post-intervention, discharge day, and postoperative day 14, with follow-up assessments at 6 weeks, 3, 6, and 12 months after discharge. The primary outcome will be the Knee Injury and Osteoarthritis Outcome Score 3 months after TKA and the medial-to-lateral knee contact force ratio. The secondary outcomes will be changes in lower-limb muscle strength, knee pain, inflammation response, active range of motion, performance-based function (5-times Sit-to-Stand Test, Time Up-and-Go Test, 3 m walking speed), Berg Balance Scale, and surgical need. DISCUSSION: Lower-limb IMS can improve muscle strength, pain, and function for early knee OA patients. However, the effects of different doses of IMS on perioperative and postoperative knee pain, inflammation response, and function after TKA are inconclusive. This study will provide high-quality evidence to assess the efficacy of IMS in end-stage patients who will undergo TKA. Understanding IMS in end-stage knee OA patients will help guide future surgical management for this population. Trial Registration Chinese Clinical Trial Registry ChiCTR2400088136.

Near-Infrared Unimolecular Chemiluminescence Probes for Deep-Tissue Imaging.

Chemiluminescence (CL) imaging has emerged as a promising optical imaging technique due to minimal background autofluorescence and being excitation-free. However, the emission of most chemiluminescent probes was concentrated in the visible light region, which limited the tissue penetration. Although some NIR chemiluminescence probes have been reported based on the chemiluminescence resonance energy transfer (CRET) strategy, the energy loss was inevitable. Thus, it is crucial to develop near-infrared (NIR) unimolecular probes with direct chemiluminescence. Herein, we propose a strategy of increasing conjugation for designing and synthesizing novel NIR chemiluminescence unimolecular probes that consist of luminol, electron acceptor, π-bridge, and electron donor. Luminol was conjugated to the unimolecular backbone to produce direct NIR chemiluminescence. Notably, the direct CL mechanism of probes was investigated. Compared with CRET-based chemiluminescence, this direct CL was more advantageous to immediately convert the chemical energy into chemiluminescence, avoiding energy degradation. Furthermore, the corresponding nanoparticles with great biosafety were prepared by self-assembly with amphiphilic DSPE-PEG. Especially, TTBL@PEG-NPs with NIR-I emission were successfully used in the sensitive in vivo chemiluminescence imaging of various inflammation models, such as peritonitis, ear swelling, and colitis. This study paves the way for the design of NIR unimolecular chemiluminescence probes and deep-tissue imaging.

Assessment of traumatic brain injury treatment guided by continuous monitoring of intracranial pressure and brain tissue oxygen partial pressure: A single-center pilot study.

Severe traumatic brain injury (TBI) is a leading cause of death and disability. Monitoring intracranial pressure (ICP) is recommended, but the data on the outcomes are conflicting. Adding continuous brain tissue oxygen partial pressure (PbtO2) monitoring may have some benefit but the OXY-TC suggested it did not improve 6-month neurological outcomes. This single-center pilot randomized controlled study aimed to evaluate whether adding PbtO2 monitoring was feasible and could improve the prognosis of severe TBI. The participants were randomized into either an ICP alone or an ICP + PbtO2 group for 7 days, with treatment protocols based on existing guidelines. Clinical parameters were collected hourly. The primary outcome was the feasibility of using PbtO2 monitoring. The secondary outcomes were 6-month survival, analyzed by the log-rank test, the 3- and 6-month Glasgow Outcome Scale (GOS) scores, compared between groups by chi-squared test. Seventy patients were included (36 ICP, 34 ICP + PbtO2). The ICP + PbtO2 group had lower mean daily ICP (13.4 vs. 18.2 mmHg, P = 0.0024) and higher mean daily cerebral perfusion pressure (82.1 vs. 74.5 mmHg, P = 0.0055). The ICP + PbtO2 group had higher 6-month survival (79.4 % vs. 55.6 %, P = 0.0337) and more favorable outcomes at 3 months (67.6 % vs. 38.9 %, P = 0.0160) and 6 months (70.6 % vs. 41.7 %, P = 0.0149). Adding PbtO2 monitoring to ICP monitoring is feasible in patients with severe TBI and could maybe improve the intermediate-term outcomes. The results will serve to design larger trials.

Understanding Cytochrome P450 Enzyme Substrate Inhibition and Prospects for Elimination Strategies.

Cytochrome P450 (CYP450) enzymes, which are widely distributed and pivotal in various biochemical reactions, catalyze diverse processes such as hydroxylation, epoxidation, dehydrogenation, dealkylation, nitrification, and bond formation. These enzymes have been applied in drug metabolism, antibiotic production, bioremediation, and fine chemical synthesis. Recent research revealed that CYP450 catalytic kinetics deviated from the classic Michaelis-Menten model. A notable substrate inhibition phenomenon that affects the catalytic efficiency of CYP450 at high substrate concentrations was identified. However, the substrate inhibition of various reactions catalyzed by CYP450 enzymes have not been comprehensively reviewed. This review describes CYP450 substrate inhibition examples and atypical Michaelis-Menten kinetic models, and provides insight into mechanisms of these enzymes. We also reviewed 3D structure and dynamics of CYP450 with substrate binding. Outline methods for alleviating substrate inhibition in CYP450 and other enzymes, including traditional fermentation approaches and protein engineering modifications. The comprehensive analysis presented in this study lays the foundation for enhancing the catalytic efficiency of CYP450 by deregulating substrate inhibition.

Validation of full-length radiograph based musculoskeletal modeling method to estimate medial and lateral knee contact forces.

BACKGROUND: Anatomical parameters of the pelvis, femur, and tibia derived from the full-length radiograph can be used to create a more accurate musculoskeletal model compared to marker-based linear scaling method. However, whether this model leads to more accurate estimations of medial knee contact force (MCF) and lateral knee contact force (LCF) than marker-based linear scaling method is still unknown. RESEARCH QUESTION: This main purpose of this study was to determine whether musculoskeletal model generated from full-length radiograph improves the estimations of MCF and LCF. METHODS: An open-source dataset including marker trajectories, ground reaction forces, in vivo knee contact forces, and full-length radiograph was used to evaluate the accuracy of full-length radiograph musculoskeletal modeling method. Subject-specific musculoskeletal models were created using anatomical parameters derived from the full-length radiograph or marker-based linear scaling methods. MCF and LCF were estimated using musculoskeletal simulations of normal walking trails. The accuracy of modeling methods was determined by comparing the estimated and in vivo measured MCF and LCF. RESULTS: Compared to the marker-based linear scaling approach, the full-length radiograph musculoskeletal modeling method exhibited decreases of 38.3 % and 41.3 % in root mean square error for MCF and LCF respectively, as well as reductions of 50.0 % and 49.3 % in mean peak errors for MCF and LCF respectively. SIGNIFICANCE: The full-length radiograph musculoskeletal modeling method provides a more accurate way to estimate MCF and LCF compared to the traditional maker-based linear scaling approach, which may contribute to understand the initiation, progression, and treatment of OA.

Stochastic lattice-based porous implant design for improving the stress transfer in unicompartmental knee arthroplasty.

BACKGROUND: Unicompartmental knee arthroplasty (UKA) has been proved to be a successful treatment for osteoarthritis patients. However, the stress shielding caused by mismatch in mechanical properties between human bones and artificial implants remains as a challenging issue. This study aimed to properly design a bionic porous tibial implant and evaluate its biomechanical effect in reconstructing stress transfer pathway after UKA surgery. METHODS: Voronoi structures with different strut sizes and porosities were designed and manufactured with Ti6Al4V through additive manufacturing and subjected to quasi-static compression tests. The Gibson-Ashby model was used to relate mechanical properties with design parameters. Subsequently, finite element models were developed for porous UKA, conventional UKA, and native knee to evaluate the biomechanical effect of tibial implant with designed structures during the stance phase. RESULTS: The internal stress distribution on the tibia plateau in the medial compartment of the porous UKA knee was found to closely resemble that of the native knee. Furthermore, the mean stress values in the medial regions of the tibial plateau of the porous UKA knee were at least 44.7% higher than that of the conventional UKA knee for all subjects during the most loading conditions. The strain shielding reduction effect of the porous UKA knee model was significant under the implant and near the load contact sites. For subject 1 to 3, the average percentages of nodes in bone preserving and building region (strain values range from 400 to 3000 µm/m) of the porous UKA knee model, ranging from 68.7 to 80.5%, were higher than that of the conventional UKA knee model, ranging from 61.6 to 68.6%. CONCLUSIONS: The comparison results indicated that the tibial implant with designed Voronoi structure offered better biomechanical functionality on the tibial plateau after UKA. Additionally, the model and associated analysis provide a well-defined design process and dependable selection criteria for design parameters of UKA implants with Voronoi structures.

Extracellular vesicles and macrophages in tumor microenvironment: Impact on cervical cancer.

Cervical cancer is a serious threat to women's health. Extracellular vesicles exist in most body fluids for communication between organisms, having different effects on the occurrence, development, angiogenesis, and metastasis of cervical cancer, and are expected to become new targets for treatment. Macrophages are natural immune systems closely linked to the development of cervical cancer. In recent years, an increasing number of studies have confirmed the role of extracellular vesicles and macrophages in the gynecologic tumor environment. This article reviews the mechanism of action and application prospects of extracellular vesicles and macrophages in the cervical cancer microenvironment. In addition, the relationship between extracellular vesicles and macrophages from different sources is described, which provides ideas for the diagnosis and treatment of cervical cancer.

Triglyceride-glucose index as a potential predictor of major adverse cardiovascular and cerebrovascular events in patients with coronary heart disease complicated with depression.

Background: Triglyceride-glucose (TyG) index is a surrogate marker of insulin resistance and metabolic abnormalities, which is closely related to the prognosis of a variety of diseases. Patients with both CHD and depression have a higher risk of major adverse cardiovascular and cerebrovascular events (MACCE) and worse outcome. TyG index may be able to predict the adverse prognosis of this special population. Methods: The retrospective cohort study involved 596 patients with both CHD and depression between June 2013 and December 2023. The primary outcome endpoint was the occurrence of MACCE, including all-cause death, stroke, MI and emergent coronary revascularization. The receiver operating characteristic (ROC) curve, Cox regression analysis, Kaplan-Meier survival analysis, and restricted cubic spline (RCS) analysis were used to assess the correlation between TyG index and MACCE risk of in patients with CHD complicated with depression. Results: With a median follow-up of 31 (15-62) months, MACCE occurred in 281(47.15%) patients. The area under the ROC curve of TyG index predicting the risk of MACCE was 0.765(0.726-0.804) (P<0.01). Patients in the high TyG index group(69.73%) had a significantly higher risk of developing MACCE than those in the low TyG index group(23.63%) (P<0.01). The multifactorial RCS model showed a nonlinear correlation (nonlinear P<0.01, overall P<0.01), with a critical value of 8.80 for the TyG index to predict the occurrence of MACCE. The TyG index was able to further improve the predictive accuracy of MACCE. Conclusions: TyG index is a potential predictor of the risk of MACCE in patients with CHD complicated with depression.

Integrated Anchored Stapling and Hierarchical Dynamics: MSICDA-Driven CREBBP Bromodomain Inhibition.

Despite recent success in the computational approaches of cyclic peptide design, current studies face challenges in modeling noncanonical amino acids and nonstandard cyclizations due to limited data. To address this challenge, we developed an integrated framework for the tailored design of stapled peptides (SPs) targeting the bromodomain of CREBBP (CREBBP-BrD). We introduce a powerful combination of anchored stapling and hierarchical molecular dynamics to design and optimize SPs by employing the MultiScale integrative conformational dynamics assessment (MSICDA) strategy, which involves an initial virtual screening of over 1.5 million SPs, followed by comprehensive simulations amounting to 154.54 µs across 5418 of instances. The MSICDA method provides a detailed and holistic stability view of peptide-protein interactions, systematically isolated optimized peptides and identified two leading candidates, DA#430 and DA#99409, characterized by their enhanced stability, optimized binding, and high affinity toward the CREBBP-BrD. In cell-free assays, DA#430 and DA#99409 exhibited 2- to 12-fold greater potency than inhibitor SGC-CBP30. Cell studies revealed higher peptide selectivity for cancerous versus normal cells over small molecules. DA#430 combined with (+)-JQ-1 showed promising synergistic effects. Our approach enables the identification of peptides with optimized binding, high affinity, and enhanced stability, leading to more precise and effective cyclic peptide design, thereby establishing MSICDA as a generalizable and transformative tool for uncovering novel targeted drug development in various therapeutic areas.

Development of a Human B7-H3-Specific Antibody with Activity against Colorectal Cancer Cells through a Synthetic Nanobody Library.

Nanobodies have emerged as promising tools in biomedicine due to their single-chain structure and inherent stability. They generally have convex paratopes, which potentially prefer different epitope sites in an antigen compared to traditional antibodies. In this study, a synthetic phage display nanobody library was constructed and used to identify nanobodies targeting a tumor-associated antigen, the human B7-H3 protein. Combining next-generation sequencing and single-clone validation, two nanobodies were identified to specifically bind B7-H3 with medium nanomolar affinities. Further characterization revealed that these two clones targeted a different epitope compared to known B7-H3-specific antibodies, which have been explored in clinical trials. Furthermore, one of the clones, dubbed as A6, exhibited potent antibody-dependent cell-mediated cytotoxicity (ADCC) against a colorectal cancer cell line with an EC50 of 0.67 nM, upon conversion to an Fc-enhanced IgG format. These findings underscore a cost-effective strategy that bypasses the lengthy immunization process, offering potential rapid access to nanobodies targeting unexplored antigenic sites.

Molecular Dynamics and In Vitro Studies Elucidating the Tunable Features of Reconfigurable Nanodiscs for Guiding the Optimal Design of Curcumin Formulation.

In this study, we advance our exploration of Apolipoprotein A-I (apoA-I) peptide analogs (APAs) for their application in nanodisc (ND) assembly, focusing on the dynamic conformational characteristics and the potential for drug delivery. We explore APA-ND interactions with an emphasis on curcumin encapsulation, utilizing molecular dynamic simulations and in vitro assessments to evaluate the efficacy of various APA-ND formulations as drug carriers. The methodological approach involved the generation of three unique apoA-I α-11/3 helical mimics, resulting in fifteen distinct APAs. Their structural integrity was rigorously assessed using ColabFold-AF2, with particular attention to pLDDT and pTM scores. Extensive molecular dynamics simulations, covering 1.7 µs across 17 ND systems, were conducted to investigate the influence of APA sequence variations on ND stability and interactions. This study reveals that the composition of APAs, notably the presence of Proline, Serine, and Tryptophan, significantly impacts ND stability and morphology. Oligomeric APAs, in particular, demonstrated superior stability and distinct interaction patterns compared to their monomeric counterparts. Additionally, hydrodynamic diameter measurements over eight weeks indicated sequence-dependent stability, highlighting the potential of specific APA configurations for sustained colloidal stability. In vitro study successfully encapsulated curcumin in [AA]3/DMPC ND formulations, revealing concentration-dependent stability and interaction dynamics. The findings underscore the remarkable capability of APA-NDs to maintain structural integrity and efficient drug encapsulation, positioning them as a promising platform for drug delivery. The study concludes by emphasizing the tunability and versatility of APA-NDs in drug formulation, potentially revolutionizing nanomedicine by enabling customized APA sequences and ND properties for targeted drug delivery.

Incidence and predictors of restenosis following successful recanalization of non-acute internal carotid artery occlusion in 252 cases.

BACKGROUND: Data concerning restenosis following successful recanalization of non-acute internal carotid artery occlusion (ICAO) are scarce. This study was conducted to identify the incidence and predictors of restenosis following successful recanalization of non-acute ICAO. METHODS: We reviewed the incidence of restenosis (defined as >70% restenosis or reocclusion) among 252 consecutive patients with successful recanalization of non-acute ICAO. Baseline, imaging, and surgery-related characteristics were analyzed to assess their association with restenosis. A scoring system was developed to identify high-risk patients for restenosis. RESULTS: During a median follow-up of 12.6 months, restenosis occurred in 56 patients (22.2%), including 39 with reocclusion and 17 with >70% restenosis. The cumulative restenosis rate was 18.0% at 12 months and 24.1% at 24 months. The incidence of stroke was higher in patients with restenosis (25.0% vs 1.5%, P<0.01). Multivariate analysis showed occlusion length (5-10 cm vs <5 cm (hazard ratio (HR) 3.15, 95% confidence interval (95% CI) 1.07 to 9.29); ≥ 10 cm vs <5 cm (HR 5.01, 95% CI 1.73 to 14.49)), residual stenosis ≥30% (HR 3.08, 95% CI 1.79 to 5.30), and internal carotid artery (ICA) wall collapse (HR 1.96, 95% CI 1.12 to 3.44) as independent predictors of restenosis. Point scores proportional to model coefficients were assigned, with scores ranging from 0 to 6. Patients scoring 3-6 had a 4.00 times higher chance of developing restenosis (95% CI 2.35 to 6.79) compared with those scoring 0-2. CONCLUSIONS: Nearly one in five patients experienced restenosis following successful recanalization of non-acute ICAO. Occlusion length, residual stenosis ≥30%, and ICA wall collapse were independently associated with restenosis.

Full-length radiograph based automatic musculoskeletal modeling using convolutional neural network.

Full-length radiographs contain information from which many anatomical parameters of the pelvis, femur, and tibia may be derived, but only a few anatomical parameters are used for musculoskeletal modeling. This study aimed to develop a fully automatic algorithm to extract anatomical parameters from full-length radiograph to generate a musculoskeletal model that is more accurate than linear scaled one. A U-Net convolutional neural network was trained to segment the pelvis, femur, and tibia from the full-length radiograph. Eight anatomic parameters (six for length and width, two for angles) were automatically extracted from the bone segmentation masks and used to generate the musculoskeletal model. Sørensen-Dice coefficient was used to quantify the consistency of automatic bone segmentation masks with manually segmented labels. Maximum distance error, root mean square (RMS) distance error and Jaccard index (JI) were used to evaluate the geometric accuracy of the automatically generated pelvis, femur and tibia models versus CT bone models. Mean Sørensen-Dice coefficients for the pelvis, femur and tibia 2D segmentation masks were 0.9898, 0.9822 and 0.9786, respectively. The algorithm-driven bone models were closer to the 3D CT bone models than the scaled generic models in geometry, with significantly lower maximum distance error (28.3 % average decrease from 24.35 mm) and RMS distance error (28.9 % average decrease from 9.55 mm) and higher JI (17.2 % average increase from 0.46) (P < 0.001). The algorithm-driven musculoskeletal modeling (107.15 ± 10.24 s) was faster than the manual process (870.07 ± 44.79 s) for the same full-length radiograph. This algorithm provides a fully automatic way to generate a musculoskeletal model from full-length radiograph that achieves an approximately 30 % reduction in distance errors, which could enable personalized musculoskeletal simulation based on full-length radiograph for large scale OA populations.

Alternative splicing-related long noncoding RNA ANRIL facilitates hepatocellular carcinoma by targeting the miR-199a-5p/SRSF1 axis and impacting Anillin.

Hepatocellular carcinoma (HCC) is characterized by aberrant alternative splicing (AS), which plays an important part in the pathological process of this disease. However, available reports about genes and mechanisms involved in AS process are limited. Our previous research has identified ANRIL as a long noncoding RNA related to the AS process of HCC. Here, we investigated the exact effect and the mechanism of ANRIL on HCC progress. The ANRIL expression profile was validated using the real-time quantitative polymerase chain reaction assay. The western blot analysis and IHC assay were conducted on candidate targets, including SRSF1 and Anillin. The clinicopathological features of 97 patients were collected and analyzed. Loss-of and gain-of-function experiments were conducted. The dual-luciferase reporter assay was applied to verify the interaction between ANRIL, miR-199a-5p, and SRSF1. Anomalous upregulation of ANRIL in HCC was observed, correlating with worse clinicopathological features of HCC. HCC cell proliferation, mobility, tumorigenesis, and metastasis were impaired by depleting ANRIL. We found that ANRIL acts as a sponger of miRNA-199a-5p, resulting in an elevated level of its target protein SRSF1. The phenotypes induced by ANRIL/miR-199a-5p/SRSF1 alteration are associated with Anillin, a validated HCC promoter. ANRIL is an AS-related lncRNA promoting HCC progress by modulating the miR-199a-5p/SRSF1 axis. The downstream effector of this axis in the development of HCC is Anillin.

Assessment of intestinal luminal stenosis and prediction of endoscopy passage in Crohn's disease patients using MRI.

BACKGROUND: Crohn's disease (CD) is an inflammatory disease of the gastrointestinal tract. The disease behavior changes over time, and endoscopy is crucial in evaluating and monitoring the course of CD. To reduce the economic burden of patients and alleviate the discomfort associated with ineffective examination, it is necessary to fully understand the location, extent, and severity of intestinal stenosis in patients with CD before endoscopy. This study aimed to utilize imaging features of magnetic resonance enterography (MRE) to evaluate intestinal stenosis in patients with CD and to predict whether endoscopy could be passed. METHODS: MRE data of patients with CD were collected, while age, gender, disease duration, and laboratory test parameters were also gathered. Two radiologists analyzed the images and assessed whether endoscopy could be passed based on the imaging performance. Imaging features of MRE were analyzed in groups based on endoscopy results. RESULTS: The readers evaluated the imaging performance for 86 patients to determine if endoscopy could be passed and performed a consistency test (compared between two readers k = 0.812, p = 0.000). In the univariate analysis, statistical differences were observed in the degree of T1WI enhancement, thickness of the intestine wall at the stenosis, and diameter of the upstream intestine between the two groups of whether endoscopy was passed. In multivariate logistic regression, the diameter of the upstream intestine was identified to be an independent factor in predicting whether endoscopy was passed or not (OR = 3.260, p = 0.046). CONCLUSIONS: The utilization of MRE signs for assessing the passage of an endoscope through the narrow segment revealed that the diameter of the upstream intestine emerged as an independent predictor of endoscopic passage. Before performing an endoscopy, MRE can aid in evaluating the passage of the endoscope. CRITICAL RELEVANCE STATEMENT: This retrospective study explored the imaging features of MRE to evaluate intestinal stenosis in patients with Crohn's disease and determined that the diameter of the upstream intestine of the stenotic segment was an independent predictor in assessing endoscopic passage. KEY POINTS: • Endoscopy is crucial in evaluating and monitoring the course of Crohn's disease. • The diameter of the upstream intestine of the stenotic segment was an independent predictor in assessing endoscopic passage. • MRE can aid in evaluating the passage of the endoscope in stenotic segments of Crohn's disease.

Effects of Long-Term Walking Exercise on Structural Progression, Symptoms, and Extensor Muscle Strength in Patients With Mild or at High Risk of Knee Osteoarthritis: Data From the Osteoarthritis Initiative.

OBJECTIVE: The aim of the study is to assess the relationship between walking exercise and medial joint space narrowing progression, symptoms, and knee extensor muscle strength in early knee osteoarthritis patients. METHODS: This nested cohort study within the Osteoarthritis Initiative included participants aged 50 yrs and older with knee osteoarthritis (Kellgren-Lawrence grades 0-2). Walking exercisers were identified using a modified Historical Physical Activity Survey Instrument. Differences in medial joint space narrowing, Knee Injury and Osteoarthritis Outcome Score, and knee extensor muscle strength were evaluated through the three-way analysis of variance. RESULTS: Among 896 participants, 83.4% reported walking exercise. Female walkers showed significant improvements in symptoms (Knee Injury and Osteoarthritis Outcome Score-pain: P < 0.001; Knee Injury and Osteoarthritis Outcome Score-symptom: P < 0.001; Knee Injury and Osteoarthritis Outcome Score-quality of life: P < 0.001; Knee Injury and Osteoarthritis Outcome Score-function, sports, and recreational activities: P = 0.007). Similar results were observed in male walkers (Knee Injury and Osteoarthritis Outcome Score-pain: P < 0.001; Knee Injury and Osteoarthritis Outcome Score-symptom: P < 0.001; Knee Injury and Osteoarthritis Outcome Score-quality of life: P = 0.001; Knee Injury and Osteoarthritis Outcome Score-function, sports, and recreational activities: P = 0.009). Walkers showed significantly increased knee extensor muscle strength at 24 mos (female: P < 0.001; male: P = 0.003). Female nonwalkers had significantly decreased knee extensor muscle strength at 24 mos ( P < 0.001). Walkers showed significant improvement in medial joint space narrowing (odds ratio = 1.1, 95% confidence interval = 1.0-1.2) and Kellgren-Lawrence grade (OR = 1.0, 95% CI = 1.0-1.1) compared with nonwalkers. CONCLUSIONS: Walking exercise prevents structural progression and improves symptoms. Meanwhile, the increased knee extensor muscle strength in all walkers further supports the validity of recommending walking exercises for early-stage knee osteoarthritis patients.

BRCA2, PALB2, RECQL4 Germline Pathogenic Variants, and Somatic TP53 Mutation in Triple Metachronous Malignancies: A Case Report and Literature Review.

Background: Multiple primary cancer (MPC) refers to the presence of more than one cancer in an individual. Triple primary malignancies are uncommon. Case: We report the case of a 50-year-old postmenopausal woman in our gynecology department, diagnosed with endometrial cancer, ovarian cancer, and unilateral breast cancer. She carried germline mutations in BRCA2, PALB2, and RECQL4, along with a somatic pathogenic variant in TP53. Endometrial cancer patients harboring germline pathogenic variants in BRCA2 exhibit a heightened risk of ovarian and breast cancer. BRCA2 is known to play a role in the development of ovarian and breast cancer, while PALB2 is identified as a gene associated with breast cancer susceptibility. RECQL4 has been linked to breast cancer, cervical cancer, and other tumors. Conclusion: Genetic testing may be imperative for identifying MPC in endometrial cancer patients. For individuals with BRCA2 and other gene pathogenic variants, routine examination and monitoring of the endometrium, ovaries, breasts, and other sites prone to polygenic cancer are recommended.

Biosynthesis of 10-Hydroxy-2-decenoic Acid through a One-Step Whole-Cell Catalysis.

10-Hydroxy-2-decenoic acid (10-HDA) is an important component of royal jelly, known for its antimicrobial, anti-inflammatory, blood pressure-lowering, and antiradiation effects. Currently, 10-HDA biosynthesis is limited by the substrate selectivity of acyl-coenzyme A dehydrogenase, which restricts the technique to a two-step process. This study aimed to develop an efficient and simplified method for synthesizing 10-HDA. In this study, ACOX from Candida tropicalis 1798, which catalyzes 10-hydroxydecanoyl coenzyme A desaturation for 10-HDA synthesis, was isolated and heterologously coexpressed with FadE, Macs, YdiI, and CYP in Escherichia coli/SK after knocking out FadB, FadJ, and FadR genes. The engineered E. coli/AKS strain achieved a 49.8% conversion of decanoic acid to 10-HDA. CYP expression was improved through ultraviolet mutagenesis and high-throughput screening, increased substrate conversion to 75.6%, and the synthesis of 10-HDA was increased to 0.628 g/L in 10 h. This is the highest conversion rate and product concentration achieved in the shortest time to date. This study provides a simple and efficient method for 10-HDA biosynthesis and offers an effective method for developing strains with high product yields.

A high-throughput molecular dynamics screening (HTMDS) approach to the design of novel cyclopeptide inhibitors of ATAD2B based on the non-canonical combinatorial library.

Cyclic peptides (CPs) are a promising class of drugs because of their high biological activity and specificity. However, the design of CP remains challenging due to their conformational flexibility and difficulties in designing stable binding conformation. Herein, we present a high-throughput MD screening (HTMDS) process for the iterative design of stable CP binders with a combinatorial CP library composed of canonical and non-canonical amino acids. As a proof of concept, we apply our methods to design CP inhibitors for the bromodomain (BrD) of ATAD2B. 698,800 CP candidates with a total of 25,570 ns MD simulations were performed to study the protein-ligand binding interactions. The binding free energies (ΔGbind) estimated by MM/PBSA approach for eight lead CP designs were found to be low. CP-1st.43 was the best CP candidate with an estimated ΔGbind of -28.48 kcal/mol when compared to the standard inhibitor C-38 which has been experimentally validated and shown to exhibit ΔGbind of -17.11 kcal/mol. The major contribution of binding sites for BrD of ATAD2B involved the hydrogen-bonding anchor within the Aly-binding pocket, salt bridging, and hydrogen-bonding mediated stabilization of the ZA loop and BC loop, and the complementary Van der Waals attraction. Our methods demonstrate encouraging results by yielding conformationally stable and high-potential CP binders that should have potential applicability in future CP drug development.Communicated by Ramaswamy H. Sarma.

A high-throughput MD screening (HTMDS) process for cyclic peptides (CPs) binders designed with canonical and non-canonical amino acids.698,800 CP candidates with a total of 25,570 ns MD simulations were performed to study the protein-ligand binding interactions and CP design.Some potent CP candidates were obtained with high binding free energies (ΔGbind) estimated by the MM/PBSA approach compared with the standard inhibitor C-38 against the bromodomain (BrD) of ATAD2B.