BEOL-Compatible 4F2 Oscillator Using Vertical InGaAs Biristor for Highly Scalable Monolithic 3D Ising Solver.

Ising solvers are important for efficiently addressing non-deterministic polynomial-time (NP)-hard combinatorial optimization problems (COPs), where scalability and compactness are crucial for practical applications. In this study, an experimental demonstration of an oscillator-based Ising solver employing a highly scalable 4F2 InGaAs bi-stable resistor (biristor) is presented. It is first explored the oscillation behavior of the InGaAs biristor, establishing that classical Ising spins can be emulated using the sub-harmonic injection locking (SHIL) technique. Furthermore, capacitive and resistive coupling between two coupled InGaAs biristors is demonstrated, leading to out-of-phase and in-phase coupling, respectively. Employing this foundational technology, it is experimentally achieved a solution to the MaxCUT problem with the InGaAs biristor-based Ising solver, supplemented by simulation-based behavior evaluations. This emerging device architecture offers a viable pathway to surmount the scaling limitations faced by present hardware-based Ising solvers, representing a significant step forward in the development of efficient, scalable solutions for complex optimization challenges.

Diagnostic accuracy of rapid antigen tests for detection of Delta and Omicron variants of severe acute respiratory syndrome coronavirus 2: Direct evidence from prospective clinical settings.

Despite widespread application during the coronavirus disease-19 pandemic, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) detection using patient-performed rapid antigen tests (RATs) is limited, especially regarding the Delta and Omicron variants. Therefore, in this study, we evaluated the performance of RATs in identifying Delta and Omicron infections in self-test settings. In this multicenter clinical performance study conducted in Korea between November 2021 and February 2022, we included participants without prior diagnostic device experience. Using 2 RAT types, we compared the results with real-time reverse transcriptase-polymerase chain reaction testing, focusing on clinical sensitivity and specificity. Reverse transcriptase-polymerase chain reaction helped confirm 77 SARS-CoV-2 infections among 280 participants. RATs exhibited high positive agreement for Omicron detection but lower rates for Delta, especially among partially vaccinated individuals. This study provides direct evidence that RATs, originally developed for ancestral strains of SARS-CoV-2, effectively detect major variants such as Delta and Omicron in real patient/clinical settings. By confirming variant presence through sequencing, our research offers significant and novel insights into the performance of RATs, particularly in the context of breakthrough infections postvaccination, with precise data on vaccination status and timing obtained from government records.

An RNA binding regulatory cascade controls the switch from proliferation to differentiation in the Drosophila male germ line stem cell lineage.

The switch from precursor cell proliferation to onset of differentiation in adult stem cell lineages must be carefully regulated to produce sufficient progeny to maintain and repair tissues, yet prevent overproliferation that may enable oncogenesis. In the Drosophila male germ cell lineage, spermatogonia produced by germ line stem cells undergo a limited number of transit amplifying mitotic divisions before switching to the spermatocyte program that sets up meiosis and eventual spermatid differentiation. The number of transit amplifying divisions is set by accumulation of the bag-of-marbles (Bam) protein to a critical threshold. In bam mutants, spermatogonia proliferate through several extra rounds of mitosis then die without becoming spermatocytes. Here we show that the key role of Bam for the mitosis to differentiation switch is repressing expression of Held Out Wings (how), homolog of mammalian Quaking. Knock down of how in germ cells was sufficient to allow spermatogonia mutant for bam or its partner benign gonial cell neoplasm (bgcn) to differentiate, while forced expression of nuclear-targeted How protein in spermatogonia wild-type for bam resulted in continued proliferation at the expense of differentiation. Our findings suggest that Bam targets how RNA for degradation by acting as an adapter to recruit the CCR4-NOT deadenylation complex via binding its subunit, Caf40. As How is itself an RNA binding protein with roles in RNA processing, our findings reveal that the switch from proliferation to meiosis and differentiation in the Drosophila male germ line adult stem cell lineage is regulated by a cascade of RNA-binding proteins.

Long-Term Results of Meniscus Allograft Transplantation With Bone Fixation Show Improved Outcomes but Progression of Joint Space Narrowing, Osteoarthritis, and Cartilage Degeneration.

PURPOSE: To investigate the clinical and objective outcomes of meniscal allograft transplantation (MAT) using bone fixation in patients after a minimum follow-up duration of 15 years and to compare the demographic factors and allograft status between patients who experienced progression of osteoarthritis and those who did not. METHODS: Consecutive patients who underwent primary MAT between December 1996 and January 2008 were reviewed retrospectively. The inclusion criterion was primary MAT with a minimum follow-up duration of 15 years. Clinical outcomes were evaluated using the modified Lysholm score, along with an evaluation of clinical failure. In objective outcomes, the progression of joint space narrowing, osteoarthritis, and the status of the associated cartilages and allografts were evaluated with follow-up radiographs and magnetic resonance imaging. RESULTS: Among the 79 cases, 54 knees in 52 patients were included in the study and evaluated for clinical outcomes. The mean Lysholm score improved from 73.9 ± 17.5 preoperatively to 86.4 ± 15.6 over a mean follow-up period of 17.5 ± 3.8 years (P < .001). Regarding minimal clinically important differences, 38 cases (70.4%) showed an improvement in the Lysholm score. The cumulative clinical survival rate was 87.0%. Objective evaluations evaluated in 32 cases with a minimum of 15 years of radiographic data revealed significant progression of joint space narrowing, osteoarthritis, and cartilage degeneration at the final follow-up, with 11 (34.4%) of 32 cases exhibiting allograft tears involving ˃50% of the allograft. Patients with progression of osteoarthritis exhibited more meniscal allograft tears and extrusion on the last follow-up magnetic resonance imaging scans than those without progression. CONCLUSIONS: Notable progression in joint space narrowing, osteoarthritis, and cartilage degeneration were observed in objective assessment. The progression of osteoarthritis might be associated with allograft tears and extrusion. In clinical evaluations, favorable long-term clinical outcomes were consistently demonstrated after MAT using the bone fixation technique. LEVEL OF EVIDENCE: Level Ⅳ, therapeutic case series.

Automated abdominal organ segmentation algorithms for non-enhanced CT for volumetry and 3D radiomics analysis.

PURPOSE: To develop fully-automated abdominal organ segmentation algorithms from non-enhanced abdominal CT and low-dose chest CT and assess their feasibility for automated CT volumetry and 3D radiomics analysis of abdominal solid organs. METHODS: Fully-automated nnU-Net-based models were developed to segment the liver, spleen, and both kidneys in non-enhanced abdominal CT, and the liver and spleen in low-dose chest CT. 105 abdominal CTs and 60 low-dose chest CTs were used for model development, and 55 abdominal CTs and 10 low-dose chest CTs for external testing. The segmentation performance for each organ was assessed using the Dice similarity coefficients, with manual segmentation results serving as the ground truth. Agreements between ground-truth measurements and model estimates of organ volume and 3D radiomics features were assessed using the Bland-Altman analysis and intraclass correlation coefficients (ICC). RESULTS: The models accurately segmented the liver, spleen, right kidney, and left kidney in abdominal CT and the liver and spleen in low-dose chest CT, showing mean Dice similarity coefficients in the external dataset of 0.968, 0.960, 0.952, and 0.958, respectively, in abdominal CT, and 0.969 and 0.960, respectively, in low-dose chest CT. The model-estimated and ground truth volumes of these organs exhibited mean differences between - 0.7% and 2.2%, with excellent agreements. The automatically extracted mean and median Hounsfield units (ICCs, 0.970-0.999 and 0.994-0.999, respectively), uniformity (ICCs, 0.985-0.998), entropy (ICCs, 0.931-0.993), elongation (ICCs, 0.978-0.992), and flatness (ICCs, 0.973-0.997) showed excellent agreement with ground truth measurements for each organ; however, skewness (ICCs, 0.210-0.831), kurtosis (ICCs, 0.053-0.933), and sphericity (ICCs, 0.368-0.819) displayed relatively low and inconsistent agreement. CONCLUSION: Our nnU-Net-based models accurately segmented abdominal solid organs in non-enhanced abdominal and low-dose chest CT, enabling reliable automated measurements of organ volume and specific 3D radiomics features.

Differences in Graft Tear Configurations in Medial and Lateral Meniscal Allograft Transplantation: Objective Evaluations Using Magnetic Resonance Imaging.

BACKGROUND: Few studies have compared the characteristics of meniscal allograft tears between medial and lateral meniscal allograft transplantation (MAT) using bone fixation. PURPOSE: To investigate the prevalence, location, and patterns of allograft tears after MAT with the bone fixation technique and compare tear patterns between medial and lateral compartments according to the time elapsed after surgery. STUDY DESIGN: Cohort study; Level of evidence, 3. METHODS: The charts of consecutive patients who underwent primary medial or lateral MAT between December 1996 and June 2019 were retrospectively reviewed. The location, pattern, and postoperative periods during which allograft tears occurred were evaluated by reviewing all series of follow-up magnetic resonance imaging (MRI) scans, which were performed at 6 weeks, 3 months, 6 months, and 1 year postoperatively and every 2 years thereafter with the patient's agreement. Postoperative periods for allograft tears were defined as the time between surgery and the follow-up MRI scan in which the meniscal tear was first confirmed. Allograft tears were compared between the medial and lateral MAT groups. RESULTS: A total of 327 consecutive patients who underwent MAT (55 medial, 272 lateral) with a minimum 2-year follow-up were retrospectively reviewed. The incidences of allograft tears after medial and lateral MATs were 32.7% and 30.9%, respectively. The mean times for tears were 80.1 ± 81.1 months and 48.9 ± 46.3 months in the medial and lateral MAT groups, respectively (P = .130). In both the medial and lateral MAT groups, allograft tears were observed mainly in the posterior horn, with complex tears being the most commonly identified tear type. In medial MATs, root tears were the second most common at 27.8%, with a significantly higher proportion than the lateral MATs (P = .014). On the other hand, in lateral MATs, meniscocapsular separation and radial tears were the second most common at 15.5% each, albeit not significantly more common than in medial MATs (P = .123 and P = .454, respectively). All root tears in medial MATs and meniscocapsular separations in lateral MATs were observed within 1 year postoperatively. CONCLUSION: Significant differences in allograft tear patterns were identified between the medial and lateral MAT groups. The proportion of root tears in medial MATs was higher than that in lateral MATs; conversely, the proportion of meniscocapsular separation was more common in lateral MATs. Such tear patterns, which may require surgical repair or graft resection, were observed only within 1 year of surgery. Therefore, close observation and regular follow-up in the earlier postoperative period are necessary after medial or lateral MATs.

Active Stratification of Colloidal Mixtures for Asymmetric Multilayers.

Stratified films offer high performance and multifunctionality, yet achieving fully stratified films remains a challenge. The layer-by-layer method, involving the sequential deposition of each layer, has been commonly utilized for stratified film fabrication. However, this approach is time-consuming, labor-intensive, and prone to leaving defects within the film. Alternatively, the self-stratification process exploiting a drying binary colloidal mixture is intensively developed recently, but it relies on strict operating conditions, typically yielding a heterogeneous interlayer. In this study, an active interfacial stratification process for creating completely stratified nanoparticle (NP) films is introduced. The technique leverages NPs with varying interfacial activity at the air-water interface. With the help of depletion pressure, the lateral compression of NP mixtures at the interface induces individual desorption of less interfacial active NPs into the subphase, while more interfacial active NPs remain at the interface. This simple compression leads to nearly perfect stratified NP films with controllability, universality, and scalability. Combined with a solvent annealing process, the active stratification process enables the fabrication of stratified films comprising a polymeric layer atop a NP layer. This work provides insightful implications for designing drug encapsulation and controlled release, as well as manufacturing transparent and flexible electrodes.

Diagnostic accuracy and predictors of alpha-synuclein accumulation in the gastrointestinal tract of Parkinson's disease.

The only characteristic of alpha-synuclein (AS) accumulation in the gastrointestinal (GI) tract of Parkinson's disease (PD) found in pathological studies is the "rostrocaudal gradient," which describes the more frequent presence of AS accumulation in the upper GI tract than in the lower GI tract. This study aimed to determine the diagnostic accuracy and identify predictors of AS accumulation in the GI tract of PD patients. The frequency of AS accumulation in the GI tract was compared between PD patients (N = 97) who underwent radical GI surgery for cancer and individually matched controls (N = 94). We evaluated AS accumulation in the neural structures using phosphorylated AS immunohistochemistry. A multivariable logistic regression analysis was conducted to determine the predictors of AS accumulation in the GI tract of PD patients. The frequency of AS accumulation was significantly higher in PD patients (75.3%) than in controls (8.5%, p-value < 0.001). The sensitivity and specificity of the full-layer evaluation were 75.3% and 91.5%, respectively. When the evaluation was confined to the mucosal/submucosal layer, the sensitivity and specificity were 46.9% and 94.7%, respectively. The rostrocaudal gradient of AS accumulation was found in PD patients. The duration from symptom onset to surgery was significantly longer in PD patients with AS accumulation (4.9 ± 4.9 years) than in PD patients without AS accumulation (1.8 ± 4.1 years, p-value = 0.005). Both disease duration and rostrocaudal gradient independently predicted the presence of AS accumulation in the GI tract of PD patients. Our study suggests PD-related AS accumulation in the GI tract follows a temporally increasing but spatially static progression pattern.

Basal Cell Carcinoma of the 2nd Digit Treated With the Reverse Cross-Finger Flap: A Case Report.

Basal cell carcinoma (BCC) is the most common form of skin cancer but rarely presents on the finger with few cases described in the literature. We present the case of a 77-year-old female with a nine-month history of biopsy-proven BCC on the dorsal aspect of her left index finger. Following the complete surgical excision of the tumor, a two-stage reverse cross-finger flap was performed, resulting in minimal impact on mobility and aesthetics and prompting a discussion on the efficacy of this approach in treating BCC of the hand. We explore the various treatment modalities available for BCCs, underscoring the importance of tailored interventions for optimal patient outcomes. The successful outcome in our case emphasizes the significance of considering alternative surgical techniques in managing uncommon presentations of BCCs, contributing to the evolving armamentarium of options available to hand surgeons.

Automated Comprehensive CT Assessment of the Risk of Diabetes and Associated Cardiometabolic Conditions.

Background CT performed for various clinical indications has the potential to predict cardiometabolic diseases. However, the predictive ability of individual CT parameters remains underexplored. Purpose To evaluate the ability of automated CT-derived markers to predict diabetes and associated cardiometabolic comorbidities. Materials and Methods This retrospective study included Korean adults (age ≥ 25 years) who underwent health screening with fluorine 18 fluorodeoxyglucose PET/CT between January 2012 and December 2015. Fully automated CT markers included visceral and subcutaneous fat, muscle, bone density, liver fat, all normalized to height (in meters squared), and aortic calcification. Predictive performance was assessed with area under the receiver operating characteristic curve (AUC) and Harrell C-index in the cross-sectional and survival analyses, respectively. Results The cross-sectional and cohort analyses included 32166 (mean age, 45 years ± 6 [SD], 28833 men) and 27 298 adults (mean age, 44 years ± 5 [SD], 24 820 men), respectively. Diabetes prevalence and incidence was 6% at baseline and 9% during the 7.3-year median follow-up, respectively. Visceral fat index showed the highest predictive performance for prevalent and incident diabetes, yielding AUC of 0.70 (95% CI: 0.68, 0.71) for men and 0.82 (95% CI: 0.78, 0.85) for women and C-index of 0.68 (95% CI: 0.67, 0.69) for men and 0.82 (95% CI: 0.77, 0.86) for women, respectively. Combining visceral fat, muscle area, liver fat fraction, and aortic calcification improved predictive performance, yielding C-indexes of 0.69 (95% CI: 0.68, 0.71) for men and 0.83 (95% CI: 0.78, 0.87) for women. The AUC for visceral fat index in identifying metabolic syndrome was 0.81 (95% CI: 0.80, 0.81) for men and 0.90 (95% CI: 0.88, 0.91) for women. CT-derived markers also identified US-diagnosed fatty liver, coronary artery calcium scores greater than 100, sarcopenia, and osteoporosis, with AUCs ranging from 0.80 to 0.95. Conclusion Automated multiorgan CT analysis identified individuals at high risk of diabetes and other cardiometabolic comorbidities. © RSNA, 2024 Supplemental material is available for this article. See also the editorial by Pickhardt in this issue.

Clinical outcomes of two-stage revision arthroplasty using a spiked tibial cement spacer in infected total knee arthroplasty.

PURPOSE: A tibial cement spacer (TCS) with spikes offers better initial stability than a conventional TCS and reduces spacer-related problems in two-stage revision total knee arthroplasty (R-TKA) for infection. We compared the clinical outcomes of two-stage revision arthroplasty for infected TKA using spiked TCS with that of conventional TCS. METHODS: This retrospective cohort study included 29 patients who underwent two-stage revision arthroplasty using an articulating cement spacer and who could be followed up for at least one year. Group S comprised 14 patients using spiked TCS, whereas Group C comprised 15 patients using conventional TCS. Demographic data, the interval from first to second stage revision, motion arc, numerical rating scale (NRS), Knee Society (KS) score, serum levels of erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP), and frequency of repeating the first-stage and infection recurrence after R-TKA between the groups were analyzed. RESULTS: No significant differences were observed in the female ratio and mean age between both groups. The mean interval between the first and second stage revision was significantly shorter in Group S than in Group C. The mean motion arc was significantly larger in Group S than in Group C. The mean NRS was significantly lower in Group S than in Group C. The mean KS score in Group S was significantly higher than that in Group C. Serum ESR and CRP levels did not differ between the groups. The frequency of repeating the first stage was lower in Group S than in Group C. However, the recurrence rate after R-TKA was higher in Group S than in Group C. CONCLUSION: Compared with conventional TCS, spiked TCS shortened the period until R-TKA and improved pain and function levels. However, no significant difference existed in the rate of infection recurrence after R-TKA.

START: A Versatile Platform for Bacterial Ligand Sensing with Programmable Performances.

Recognition of signaling molecules for coordinated regulation of target genes is a fundamental process for biological systems. Cells often rely on transcription factors to accomplish these intricate tasks, yet the subtle conformational changes of protein structures, coupled with the complexity of intertwined protein interaction networks, pose challenges for repurposing these for bioengineering applications. This study introduces a novel platform for ligand-responsive gene regulation, termed START (Synthetic Trans-Acting Riboswitch with Triggering RNA). Inspired by the bacterial ligand sensing system, riboswitch, and the synthetic gene regulator, toehold switch, the START platform enables the implementation of synthetic biosensors for various ligands. Rational sequence design with targeted domain optimization yields high-performance STARTs with a dynamic range up to 67.29-fold and a tunable ligand sensitivity, providing a simple and intuitive strategy for sensor engineering. The START platform also exhibits modularity and composability to allow flexible genetic circuit construction, enabling seamless implementation of OR, AND, and NOT Boolean logic gates for multiple ligand inputs. The START design principle is capable of broadening the suite of synthetic biosensors for diverse chemical and protein ligands, providing a novel riboregulator chassis for synthetic biology and bioengineering applications.

Preoperative Medial Meniscus Extrusion on Magnetic Resonance Imaging Is Associated With a Tendency Toward Varus Shifting of Open-Wedge High Tibial Osteotomy: A Minimum 5-Year Follow-Up Study.

PURPOSE: To investigate serial changes in postoperative alignment over 5 years after open-wedge high tibial osteotomy (OWHTO) and to identify risk factors associated with alterations in the postoperative weightbearing line (WBL) ratio. METHODS: Patients who underwent OWHTO during 2011-2017 were retrospectively reviewed. The inclusion criteria were (1) follow-up duration ≥5 years and (2) serial postoperative longstanding hip-to-ankle radiographs to evaluate alignment alterations. The WBL ratio was measured preoperatively and at 3 months, 6 months, 1 year, and 5 years postoperatively to evaluate serial changes. Alterations in the WBL ratio were analyzed using a linear mixed model, considering potential risk factors including International Cartilage Repair Society grades of each compartment and medial meniscus extrusion (≥3 mm). Clinical outcomes were assessed using the Knee Society objective and functional scores, and the correlations between clinical outcomes and alignment alteration were examined. RESULTS: A total of 78 knees were investigated. During the study period, the overall WBL ratio decreased by 5.5% ± 7.2%, signifying varus shifting, from 58.6% ± 11.5% at 3 months postsurgery to 51.5% ± 12.7% at 5 years postsurgery. Based on univariate regression analyses, International Cartilage Repair Society grades of the medial compartment and medial meniscus extrusion were included in a linear mixed model regarding alignment alteration. The model identified medial meniscus extrusion as a significant risk factor after adjusting for time (P < .001). Medial meniscus extrusion also had a significant interaction with time (P < .001), indicating greater alignment alteration in cases of medial meniscus extrusion. The extrusion was noted in 68 of 78 knees. CONCLUSIONS: In the midterm following OWHTO, the overall alignment had a tendency toward varus shifting. A linear mixed model found that preoperative medial meniscus extrusion on magnetic resonance imaging is associated with the tendency. LEVEL OF EVIDENCE: Level III, retrospective cohort study.

Induction of Diabetes Mellitus Using Alloxan in Sprague Dawley Rats.

PURPOSE: The use of rodent models for diabetes, particularly with pancreatic islet transplantation, has been prevalent in various preclinical trials. The purpose of this study is to establish a diabetes mellitus (DM) model in Sprague Dawley (SD) rats using alloxan evaluated by assessing alloxan dosage, the induction rate of diabetes, and glucose stability through insulin treatment. METHODS: Over the course of 13 experimental rounds, diabetes was induced in 86 SD rats using alloxan at concentrations of 200 mg/kg (16 rats) or 150 mg/kg (70 rats). Various parameters, including diabetes induction rates, average insulin doses, extent of weight loss, and adverse effects such as diabetic ketoacidosis (DKA), were measured. RESULTS: The administration of 200 mg/kg of alloxan in rats resulted in severe diabetes induction, leading to DKA in three individuals, despite daily insulin glargine administration, DKA prevention was unsuccessful. The stability of alloxan decreases over time, especially when refrigeration is compromised during weighing. In the group treated with 150 mg/kg of alloxan, the diabetes induction rate was 83%. The average insulin dose was 2.21 units/kg/day. In contrast, the group treated with 200 mg/kg of alloxan exhibited a diabetes induction rate of 81% with a statistically significant higher average insulin requirement at 7.58 units/kg/day compared to 150 mg/kg of alloxan. CONCLUSION: Inducing diabetes in rats with 150 mg/kg of alloxan is considered more suitable for creating a diabetes model for xenogeneic islet transplantation compared to using 200 mg/kg of alloxan. This is due to fewer complications related to DKA or hyperglycemia and reduced need for exogenous insulin treatment.

Clinical feasibility of deep learning based synthetic contrast enhanced abdominal CT in patients undergoing non enhanced CT scans.

Our objective was to develop and evaluate the clinical feasibility of deep-learning-based synthetic contrast-enhanced computed tomography (DL-SynCCT) in patients designated for nonenhanced CT (NECT). We proposed a weakly supervised learning with the utilization of virtual non-contrast CT (VNC) for the development of DL-SynCCT. Training and internal validations were performed with 2202 pairs of retrospectively collected contrast-enhanced CT (CECT) images with the corresponding VNC images acquired from dual-energy CT. Clinical validation was performed using an external validation set including 398 patients designated for true nonenhanced CT (NECT), from multiple vendors at three institutes. Detection of lesions was performed by three radiologists with only NECT in the first session and an additionally provided DL-SynCCT in the second session. The mean peak signal-to-noise ratio (PSNR) and structural similarity index map (SSIM) of the DL-SynCCT compared to CECT were 43.25 ± 0.41 and 0.92 ± 0.01, respectively. With DL-SynCCT, the pooled sensitivity for lesion detection (72.0% to 76.4%, P < 0.001) and level of diagnostic confidence (3.0 to 3.6, P < 0.001) significantly increased. In conclusion, DL-SynCCT generated by weakly supervised learning showed significant benefit in terms of sensitivity in detecting abnormal findings when added to NECT in patients designated for nonenhanced CT scans.

Effect of joint-line obliquity on long-term survivorship of total knee arthroplasty: A postoperative phenotype analysis.

PURPOSE: Whether the longevity of total knee arthroplasty (TKA) differs between postoperative phenotypes has not been investigated. This study aims to examine which phenotype has a worse long-term survival rate than the reference phenotype (neutral alignment-parallel joint line), and whether joint-line obliquity (JLO) affects the survivorship of TKA. METHODS: A total of 945 knees that underwent primary TKAs for primary osteoarthritis from January 2000 to January 2009 were included. These were classified into nine postoperative phenotypes based on the combined assessment of the hip-knee-ankle (HKA) angle and JLO, measured on standing radiographs. The 5-, 10- and 15-year survival rates were analysed using Kaplan-Meier methods and log-rank tests. The long-term survival rates of each phenotype were compared with the reference phenotype. RESULTS: There were 55 aseptic mechanical failures within a period of 10.4 ± 5.0 years. The most frequently observed phenotypes were the reference phenotype (n = 527), neutral alignment-lateral joint-line inclination (n = 162), varus alignment-lateral joint-line inclination (n = 104) and varus alignment-parallel joint line (n = 101). The overall failure rate for each phenotype was 3.6%, 3.7%, 18.3% and 7.9%, respectively. Only the 10- and 15-year survival rates of the varus alignment-lateral joint-line inclination phenotype were significantly different from those of the reference phenotype (97%-93% vs. 90%-69%; p = 0.017, <0.001). CONCLUSION: The lateral joint-line inclination phenotype had an inferior long-term survival rate after varus-aligned TKA. This suggested that alignment and JLO affected the long-term survival rate of patients who underwent TKA. LEVEL OF EVIDENCE: Level III, Retrospective cohort study.

Logical regulation of endogenous gene expression using programmable, multi-input processing CRISPR guide RNAs.

The CRISPR-Cas system provides a versatile RNA-guided approach for a broad range of applications. Thanks to advances in RNA synthetic biology, the engineering of guide RNAs (gRNAs) has enabled the conditional control of the CRISPR-Cas system. However, achieving precise regulation of the CRISPR-Cas system for efficient modulation of internal metabolic processes remains challenging. In this work, we developed a robust dCas9 regulator with engineered conditional gRNAs to enable tight control of endogenous genes. Our conditional gRNAs in Escherichia coli can control gene expression upon specific interaction with trigger RNAs with a dynamic range as high as 130-fold, evaluating up to a three-input logic A OR (B AND C). The conditional gRNA-mediated targeting of endogenous metabolic genes, lacZ, malT and poxB, caused differential regulation of growth in Escherichia coli via metabolic flux control. Further, conditional gRNAs could regulate essential cytoskeleton genes, ftsZ and mreB, to control cell filamentation and division. Finally, three types of two-input logic gates could be applied for the conditional control of ftsZ regulation, resulting in morphological changes. The successful operation and application of conditional gRNAs based on programmable RNA interactions suggests that our system could be compatible with other Cas-effectors and implemented in other host organisms.

Melatonin alleviates myocardial dysfunction through inhibition of endothelial-to-mesenchymal transition via the NF-κB pathway.

Endothelial-to-mesenchymal transition (EndMT) is a complex biological process of cellular transdifferentiation by which endothelial cells (ECs) lose their characteristics and acquire mesenchymal properties, leading to cardiovascular remodeling and complications in the adult cardiovascular diseases environment. Melatonin is involved in numerous physiological and pathological processes, including aging, and has anti-inflammatory and antioxidant activities. This molecule is an effective therapeutic candidate for preventing oxidative stress, regulating endothelial function, and maintaining the EndMT balance to provide cardiovascular protection. Although recent studies have documented improved cardiac function by melatonin, the mechanism of action of melatonin on EndMT remains unclear. The present study investigated the effects of melatonin on induced EndMT by transforming growth factor-ß2/interleukin-1ß in both in vivo and in vitro models. The results revealed that melatonin reduced the migratory ability and reactive oxygen species levels of the cells and ameliorated mitochondrial dysfunction in vitro. Our findings indicate that melatonin prevents endothelial dysfunction and inhibits EndMT by activating related pathways, including nuclear factor kappa B and Smad. We also demonstrated that this molecule plays a crucial role in restoring cardiac function by regulating the EndMT process in the ischemic myocardial condition, both in vessel organoids and myocardial infarction (MI) animal models. In conclusion, melatonin is a promising agent that attenuates EC dysfunction and ameliorates cardiac damage compromising the EndMT process after MI.

Reconstruction of Segmental Bone Defect in Canine Tibia Model Utilizing Bi-Phasic Scaffold: Pilot Study.

The reunion and restoration of large segmental bone defects pose significant clinical challenges. Conventional strategies primarily involve the combination of bone scaffolds with seeded cells and/or growth factors to regulate osteogenesis and angiogenesis. However, these therapies face inherent issues related to immunogenicity, tumorigenesis, bioactivity, and off-the-shelf transplantation. The biogenic micro-environment created by implanted bone grafts plays a crucial role in initiating the bone regeneration cascade. To address this, a highly porous bi-phasic ceramic synthetic bone graft, composed of hydroxyapatite (HA) and alumina (Al), was developed. This graft was employed to repair critical segmental defects, involving the creation of a 2 cm segmental defect in a canine tibia. The assessment of bone regeneration within the synthetic bone graft post-healing was conducted using scintigraphy, micro-CT, histology, and dynamic histomorphometry. The technique yielded pore sizes in the range of 230-430 µm as primary pores, 40-70 µm as secondary inner microchannels, and 200-400 nm as tertiary submicron surface holes. These three components are designed to mimic trabecular bone networks and to provide body fluid adsorption, diffusion, a nutritional supply, communication around the cells, and cell anchorage. The overall porosity was measured at 82.61 ± 1.28%. Both micro-CT imaging and histological analysis provided substantial evidence of robust bone formation and the successful reunion of the critical defect. Furthermore, an histology revealed the presence of vascularization within the newly formed bone area, clearly demonstrating trabecular and cortical bone formation at the 8-week mark post-implantation.