In Vitro Performance of a Long-Range Surface Plasmon Hydrogel Aptasensor for Continuous and Real-Time Vancomycin Measurement in Human Serum.

This study investigated the suitability of surface modification for a long-range surface plasmon (LRSP) aptasensor using two different hydrogels, aiming at real-time monitoring of vancomycin (VCM) in undiluted serum and blood. Three different layer structures were formed on a gold surface of LRSP sensor chip using poly[2-methacryloyloxyethyl phosphorylcholine (MPC)-co-N-methacryloyl-(L)-tyrosinemethylester (MAT)] (PMM) and poly[MPC-co-2-ethylhexyl methacrylate (EHMA)-co-MAT] (PMEM). The peptide aptamer for VCM was immobilized in PMM and PMEM via MAT. Among four differently prepared sensor chips, the LRSP hydrogel aptasensor with PMM, referred to as the PMM hydrogel, exhibited the highest sensor output and superior antifouling properties. Following the optimization of the PMM hydrogel preparation conditions, the shelf life of the PMM hydrogel was determined to exceed 2 weeks, and the same sensor chip could be used for 102 days without significant performance deterioration. The PMM hydrogel was then applied for VCM measurement in undiluted serum in vitro, where it demonstrated a limit of detection of 0.098 µM and a dynamic range of 0.18-100 µM, covering the therapeutic range. Additionally, the PMM hydrogel enabled the continuous measurement of various VCM concentrations in serum without rinsing and showed a concentration-dependent output in undiluted blood. These findings underscore the potential of the PMM hydrogel for real-time and direct monitoring of VCM in body fluids.

Intraoperative reliability of the tibial anteroposterior axis "Akagi's Line" in total knee arthroplasty.

Purpose: The tibial anatomical anteroposterior (AP) axis "Akagi's line" was originally defined on computed tomography (CT) in total knee arthroplasty (TKA); however, its intraoperative reproducibility remains unknown. This study aimed to evaluate the intraoperative reproducibility of the Akagi's line and its effect on postoperative clinical outcomes. Methods: This prospective study included 171 TKAs. The rotational angle of the intraoperative Akagi's line relative to the original Akagi's line (RAA) defined on CT was measured. The RAA was calculated based on the tibial component rotational angles relative to the intraoperative Akagi's line measured using the navigation system and CT. The effects of RAA on postoperative clinical outcomes and rotational alignments of components were also evaluated. Results: The mean absolute RAA (standard deviation) value was 5.5° (3.9°). The range of RAA was 22° internal rotation to 16° external rotation. Intraoperative Akagi's line outliers (RAA > 10°) were observed in 14% of the knees (24 knees). In outlier analysis, the tibial component rotation angle was externally rotated 6.5° (5.6°) in the outlier group and externally rotated 3.7° (4.2°) in the nonoutlier group (≤10°), with a significant difference between the two groups. Additionally, the outlier group (RAA > 10°) showed lower postoperative clinical outcomes. Conclusion: The original Akagi's line defined on CT showed insufficient reproducibility intraoperatively. The poor intraoperative detection of Akagi's line could be the reason for the tibial component rotational error and worse postoperative clinical outcomes. Level of Evidence: Level IV, case series.

Mouthguard-Type Wearable Sensor for Monitoring Salivary Turbidity to Assess Oral Hygiene.

Salivary turbidity is a promising indicator for evaluating oral hygiene. This study proposed a wearable mouthguard-type sensor for continuous and unconstrained measurement of salivary turbidity. The sensor evaluated turbidity by measuring the light transmittance of saliva with an LED and a phototransistor sealed inside a double-layered mouthguard. The sensor was also embedded with a Bluetooth wireless module, enabling the wireless measurement of turbidity. The mouthguard materials (polyethylene terephthalate-glycol and ethylene-vinyl acetate) and the wavelength of the LED (405 nm) were experimentally determined to achieve high sensitivity in salivary turbidity measurement. The turbidity quantification characteristic of the proposed sensor was evaluated using a turbidity standard solution, and the sensor was capable of turbidity quantification over a wide dynamic range of 1-4000 FTU (formazine turbidity unit), including reported salivary turbidity (400-800 FTU). In vitro turbidity measurement using a saliva sample showed 553 FTU, which is equivalent to the same sample measured with a spectrophotometer (576 FTU). Moreover, in vivo experiments also showed results equivalent to that measured with a spectrophotometer, and wireless measurement of salivary turbidity was realized using the mouthguard-type sensor. Based on these results, the proposed mouthguard-type sensor has promising potential for the unconstrained continuous evaluation of oral hygiene.

Factors Associated With a Lateral Ankle Sprain in Young Female Soccer Players: A Prospective Cohort Study.

Background: Previous studies have attempted to determine if certain risk factors can predict the occurrence of a lateral ankle sprain (LAS) in female soccer players. Unfortunately, there is limited evidence with regard to risk factors associated with an LAS in female soccer players. Purpose: To identify intrinsic risk factors for an LAS among young female soccer players. Study Design: Cohort study; Level of evidence, 2. Methods: Participants were 161 young female soccer players in Japan who were evaluated for LAS risk factors during a preseason medical assessment. The assessment included anthropometric, joint laxity, joint range of motion, muscle flexibility, muscle strength, and balance measurements. Each athlete's history of LASs was also collected. The participants were monitored during a single-yearseason for LASs, as diagnosed by physicians. Results: There were 26 instances of an LAS in 25 players (15.5%) during the season. Injured players were significantly more likely to have sustained a previous ankle sprain (P = .045) and demonstrated significantly worse balance than their peers without an LAS during the double- and single-leg balance tests (P = .008 for both). Athletes with lower hamstring-to-quadriceps muscle strength ratios were also significantly more likely to sustain an LAS (P = .02). Conclusion: Poor balance, a low hamstring-to-quadriceps ratio, and a history of ankle sprains were associated with an increased risk of LASs in young female soccer players in the current study. These findings may be useful for developing a program to prevent LASs in this population.

Lower hamstring to quadriceps muscle strength ratio and lower body weight as factors associated with noncontact anterior cruciate ligament injury in male American football players: A prospective cohort study.

Background: Previous studies have aimed to determine the use of certain risk factors in predicting the occurrence of noncontact anterior cruciate ligament (ACL) injuries. Unfortunately, evidence regarding noncontact ACL injuries in male American football players is limited. This prospective cohort study aimed to identify intrinsic risk factors for noncontact ACL injury among male American football players. Methods: This study evaluated 152 male American football players in Japan for potential noncontact ACL injury risk factors during a preseason medical assessment, including anthropometric, joint laxity, and flexibility, muscle flexibility, muscle strength, and balance measurements. A total of 25 variables were examined. Participants were monitored during each season for noncontact ACL injury, as diagnosed by physicians. Results: Noncontact ACL injuries occurred in 11 knees of 11 players (prevalence; 7.1 %). Injured players were significantly more likely to have lightweight (P = 0.049). No statistically significant between-group differences were found for any other variables. Participants with a lower hamstring to quadriceps (H/Q) ratio (P = 0.04) were more likely to sustain noncontact ACL injuries. Conclusion: Lower H/Q ratio and lower body weight were significantly associated with new-onset noncontact ACL injury in male American football players. These findings will help develop strategies to prevent noncontact ACL injuries in male American football players.

Postoperative quadriceps weakness and male sex are risk factors for patellofemoral articular cartilage lesions after anatomical anterior cruciate ligament reconstruction.

PURPOSE: Patellofemoral (PF) compartment cartilage lesions are a frequent problem after anterior cruciate ligament (ACL) reconstruction. This study aimed to determine the factors that influence PF cartilage lesions after anatomical ACL reconstruction. METHODS: This study enrolled a total of 114 patients who did not manifest PF compartment cartilage lesions during anatomical ACL reconstruction and underwent second-look arthroscopy 18 months postoperatively. Arthroscopy using the International Cartilage Repair Society (ICRS) classification was used to assess cartilage lesions. The correlation between surgical findings, radiographic factors, and clinical factors and change of ICRS grade was analysed. Multivariate regression analysis was conducted to reveal the independent risk factors for PF cartilage lesions among patients' demographic data and parameters that correlated with the change of ICRS grade in the correlation analyses. RESULTS: ICRS grade changes in PF cartilage were significantly correlated with age, sex, quadriceps strength at 1 year postoperatively, hamstrings strength at pre- and 1 year postoperatively, and single leg hop test at 1 year postoperatively. However, no significant correlation was found between the time between injury and surgery, posterior tibial slope angle, pre- and postoperative Tegner activity scale, graft type, initial graft tension, meniscus injury, meniscus injury treatment, pre- and postoperative range of motion, anteroposterior laxity and preoperative quadriceps strength, and the change in ICRS grade. Multivariate regression analysis revealed male (P = 0.019) and quadriceps strength weakness at 1 year postoperatively (P = 0.009) as independent risk factors for PF cartilage lesions. CONCLUSIONS: Quadriceps strength weakness 1 year after ACL reconstruction and males were correlated with a new PF cartilage lesion after anatomical ACL reconstruction, with no significant correlation between bone-patellar tendon-bone autograft, initial graft tension, or extension deficit and new PF cartilage lesion. Rehabilitation that focuses on quadriceps strength after ACL reconstruction is recommended to prevent new PF cartilage lesions. LEVEL OF EVIDENCE: Level IV.

Comparison of finite helical axis of the knee joint before and after bi-cruciate-preserving knee arthroplasty.

BACKGROUND: Difference in the finite helical axis of the knee joints before and after anterior cruciate ligament-preserving knee arthroplasties such as unicompartmental knee arthroplasty and bi-cruciate-retaining total knee arthroplasty remains unknown. This study compared the knee finite helical axes before and after bi-cruciate-preserving knee arthroplasty. METHODS: Patients undergoing medial unicompartmental knee arthroplasty and bi-cruciate-retaining total knee arthroplasty were included. Under fluoroscopy, participants performed a deep knee bend before and after surgery. A two/three-dimensional registration technique was employed to measure tibiofemoral kinematics. Femoral finite helical axis was calculated in a flexion range of 0-120° using 30° windows (early-, mid-, late-, and deep-flexion phases). FINDINGS: In unicompartmental knee arthroplasty, the preoperative knee vertical angle was larger than the postoperative vertical angle in mid- and deep-flexion phases. The postoperative knee vertical angle was smaller in unicompartmental knee arthroplasty than in bi-cruciate-retaining total knee arthroplasty. In unicompartmental knee arthroplasty, the preoperative horizontal angle was smaller than the postoperative horizontal angle in the early-flexion phase. However, in bi-cruciate-retaining total knee arthroplasty, the preoperative horizontal angle was larger than the postoperative horizontal angle in mid- and deep-flexion phases. The horizontal angle was smaller before unicompartmental knee arthroplasty than that before bi-cruciate-retaining total knee arthroplasty in early-, mid-, and deep-flexion phases. However, the vertical angle was larger after unicompartmental knee arthroplasty than that after bi-cruciate-retaining total knee arthroplasty in the early-flexion phase. INTERPRETATION: The knee finite helical axes before and after unicompartmental knee arthroplasty differed from those before and after bi-cruciate-retaining total knee arthroplasty.

Gas-Phase Biosensors (Bio-Sniffers) for Measurement of 2-Nonenal, the Causative Volatile Molecule of Human Aging-Related Body Odor.

The molecule 2-nonenal is renowned as the origin of unpleasant human aging-related body odor that can potentially indicate age-related metabolic changes. Most 2-nonenal measurements rely on chromatographic analytical systems, which pose challenges in terms of daily usage and the ability to track changes in concentration over time. In this study, we have developed liquid- and gas-phase biosensors (bio-sniffers) with the aim of enabling facile and continuous measurement of trans-2-nonenal vapor. Initially, we compared two types of nicotinamide adenine dinucleotide (phosphate) [NAD(P)]-dependent enzymes that have the catalytic ability of trans-2-nonenal: aldehyde dehydrogenase (ALDH) and enone reductase 1 (ER1). The developed sensor quantified the trans-2-nonanal concentration by measuring fluorescence (excitation: 340 nm, emission: 490 nm) emitted from NAD(P)H that was generated or consumed by ALDH or ER1. The ALDH biosensor reacted to a variety of aldehydes including trans-2-nonenal, whereas the ER1 biosensor showed high selectivity. In contrast, the ALDH bio-sniffer showed quantitative characteristics for trans-2-nonenal vapor at a concentration range of 0.4-7.5 ppm (with a theoretical limit of detection (LOD) and limit of quantification (LOQ) of 0.23 and 0.26 ppm, respectively), including a reported concentration (0.85-4.35 ppm), whereas the ER1 bio-sniffer detected only 0.4 and 0.8 ppm. Based on these findings, headspace gas of skin-wiped alcohol-absorbed cotton collected from study participants in their 20s and 50s was measured by the ALDH bio-sniffer. Consequently, age-related differences in signals were observed, suggesting the potential for measuring trans-2-nonenal vapor.

Real-Time Continuous Monitoring of Oral Soft Tissue Pressure with a Wireless Mouthguard Device for Assessing Tongue Thrusting Habits.

In orthodontics, understanding the pressure of oral soft tissues on teeth is important to elucidate the cause and establish treatment methods. We developed a small wireless mouthguard (MG)-type device that continuously and unrestrainedly measures pressure, which had previously been unachieved, and evaluated its feasibility in human subjects. First, the optimal device components were considered. Next, the devices were compared with wired-type systems. Subsequently, the devices were fabricated for human testing to measure tongue pressure during swallowing. The highest sensitivity (51-510 g/cm2) with minimum error (CV < 5%) was obtained using an MG device with polyethylene terephthalate glycol and ethylene vinyl acetate for the lower and upper layers, respectively, and with a 4 mm PMMA plate. A high correlation coefficient (0.969) was observed between the wired and wireless devices. In the measurements of tongue pressure on teeth during swallowing, 132.14 ± 21.37 g/cm2 for normal and 201.17 ± 38.12 g/cm2 for simulated tongue thrust were found to be significantly different using a t-test (n = 50, p = 6.2 × 10-19), which is consistent with the results of a previous study. This device can contribute to assessing tongue thrusting habits. In the future, this device is expected to measure changes in the pressure exerted on teeth during daily life.

xcore: an R package for inference of gene expression regulators.

BACKGROUND: Elucidating the Transcription Factors (TFs) that drive the gene expression changes in a given experiment is a common question asked by researchers. The existing methods rely on the predicted Transcription Factor Binding Site (TFBS) to model the changes in the motif activity. Such methods only work for TFs that have a motif and assume the TF binding profile is the same in all cell types. RESULTS: Given the wealth of the ChIP-seq data available for a wide range of the TFs in various cell types, we propose that gene expression modeling can be done using ChIP-seq "signatures" directly, effectively skipping the motif finding and TFBS prediction steps. We present xcore, an R package that allows TF activity modeling based on ChIP-seq signatures and the user's gene expression data. We also provide xcoredata a companion data package that provides a collection of preprocessed ChIP-seq signatures. We demonstrate that xcore leads to biologically relevant predictions using transforming growth factor beta induced epithelial-mesenchymal transition time-courses, rinderpest infection time-courses, and embryonic stem cells differentiated to cardiomyocytes time-course profiled with Cap Analysis Gene Expression. CONCLUSIONS: xcore provides a simple analytical framework for gene expression modeling using linear models that can be easily incorporated into differential expression analysis pipelines. Taking advantage of public ChIP-seq databases, xcore can identify meaningful molecular signatures and relevant ChIP-seq experiments.

Intraoperative femoral rotational kinematics are similar in varus and valgus knees during medial pivot total knee arthroplasty.

BACKGROUND: Medial-pivot type total knee arthroplasty is designed to reproduce physiological femoral rotational kinematics during knee flexion; however, its rotational kinematics in valgus knees remain unknown. This study's hypothesis is that its kinematics show the similar medial pivot motion in valgus knees as in varus knees. METHODS: This retrospective study included 50 cases of primary medial-pivot type total knee arthroplasty performed with navigation for knee osteoarthritis. Cases were grouped as valgus (n = 20) or varus (n = 30). In valgus knees, surgeons used preoperative manual testing to confirm that alignment was correctable. They evaluated femoral rotational kinematics at maximum extension, 30°, 60°, 90°, and maximum flexion, using a navigation system, both before and after implantation. Finally, intraoperative rotational kinematics, postoperative patient-reported outcomes, and knee range of motion were compared between the two groups. FINDINGS: Before implantation, both valgus and varus knees displayed external femoral rotation relative to the tibia during knee flexion. The rotation magnitude was significantly larger in varus knees at 60°, 90°, and maximum flexion angles compared to valgus knees (P < 0.05). In contrast, after implantation, both groups displayed external femoral rotation of 12-13° during knee flexion with no significant differences between the two groups at any knee flexion angle tested (P > 0.05). Short-term clinical outcomes were comparable between valgus and varus knees. INTERPRETATION: Postoperatively, medial-pivot type total knee arthroplasty for knee osteoarthritis resulted in external femoral rotation during knee flexion. Notably, this occurred even in valgus knees with good short-term clinical outcomes.

Anterior cruciate ligament-deficient knee induces a posterior location of the femur in the medial compartment during squatting.

Although an anterior cruciate ligament (ACL) deficiency induces knee osteoarthritis, particularly in the medial compartment, the kinematics remains partially unclear. This study investigated the in vivo knee kinematics of ACL-deficient and normal knees by comparing them during squatting. This prospective comparative study included 17 ACL-deficient knees and 20 normal knees. The kinematics was investigated under fluoroscopy using a two- to three-dimensional registration technique. The anteroposterior (AP) translation of the medial and lateral sides of the femur, axial rotation of the femur relative to the tibia, and kinematic pathways were evaluated and compared. At first, the medial AP position of the femur translated anteriorly from 0° to midflexion, afterward it translated posteriorly in both ACL-deficient and normal knees. However, the medial AP position of the femur in the ACL-deficient knees was located significantly posteriorly compared with normal knees at 0-110° flexion. The lateral AP position of the femur translated posteriorly from 0° to 150° flexion in both ACL-deficient and normal knees. The lateral AP position of the femur in the ACL-deficient knees was located significantly posteriorly compared with that in normal knees at 0-10° flexion. The femur showed external rotation from 0° to 150° flexion in both ACL-deficient and normal knees. A medial pivot motion and subsequent bicondylar rollback were observed in both knees in the kinematic pathway. In conclusion, the AP position of the femur relative to the tibia was altered due to ACL deficiency, particularly in the medial compartment.

Long-range surface plasmon aptasensor for label-free monitoring of vancomycin.

Vancomycin (VCM) causes poisoning symptoms at high concentrations; thus, therapeutic drug monitoring is recommended to measure and control blood levels regularly. However, blood analysis at regular intervals does not allow knowing the detailed temporal change in concentration. To address this challenge, we developed a long-range surface plasmon (LRSP) aptasensor for measuring VCM label-free and real-time by combining a sensitive LRSP sensor and a peptide aptamer with a VCM recognition site. First, three different biosensors for VCM were compared. One was prepared by immobilizing the peptide aptamer directly on (Direct-Apt) or via a self-assembled monolayer (SAM) on a gold surface (SAM-Apt). The other used anti-VCM antibodies immobilized on a gold surface via the SAM (SAM-Ab). The Direct-Apt showed larger sensor output to VCM than the other biosensors. The dynamic range for VCM was 0.78-100 µM, including the therapeutic range (6.9-13.8 µM). The Direct-Apt also showed the sensor output only from VCM among four different antibiotics, demonstrating the high selectivity for VCM. The VCM captured by the aptamer could be removed by rinsing with phosphate-buffered saline. The measurement was rapid, with 72- and 77-sec response and recovery times, allowing not only repeated but also real-time measurements. Finally, the Direct-Apt in 20% serum solutions showed comparable sensitivity to VCM in the buffer solution, indicating high capability for real-sample.

In vivo kinematic comparison of medial pivot total knee arthroplasty in weight-bearing and non-weight-bearing deep knee bending.

BACKGROUND: This study aimed to determine the kinematics of medial pivot total knee arthroplasty by comparing weight-bearing and non-weight-bearing deep knee bending and to evaluate the effect of the weight-bearing state on the kinematics. METHODS: The kinematics of 19 knees were investigated under fluoroscopy during squatting (weight-bearing) and active-assisted knee bending (non-weight-bearing) using two- to three-dimensional registration technique. Accordingly, range of motion, anteroposterior translation for the medial and lateral low contact points, axial rotation of the femoral component relative to the tibial component and kinematic pathway were evaluated. FINDINGS: There was no difference in range of motion between the two states. The medial anteroposterior translation showed no significant movement with no anterior translation in both the weight-bearing and non-weight-bearing from 0° to 90° of flexion. Regarding the lateral anteroposterior translation, a posterior translation was observed during weight-bearing, whereas a slight anterior translation from 0° to 30° of flexion and subsequent posterior translation were found in the non-weight-bearing. Femoral external rotation was observed in the weight-bearing, whereas femoral internal rotation was seen from 0° to 30° of flexion and subsequent femoral external rotation was observed in the non-weight-bearing. The kinematic pathway showed medial pivot motion and subsequent bicondylar rollback in the weight-bearing, whereas only medial pivot motion was observed in the non-weight-bearing. INTERPRETATION: The medial anteroposterior translation of the femur during deep knee bending showed no anterior motion in the two states. The lateral anteroposterior translation and femoral rotation were different in the mid-flexion range between the two states.

Blood sorbitol measurement in diabetic rats treated with an aldose reductase inhibitor using an improved fiber-optic sorbitol biosensor.

Sorbitol is known as a biomarker for the evaluation of the progress of diabetic complications. We have developed a sorbitol biosensor using an optical fiber for rapid diagnosis and pathological evaluation of diabetic complications. In this paper, we measured blood sorbitol in diabetic rats using an improved biosensor, and discussed the effectiveness of the developed biosensor and the significance of sorbitol measurement. In order to investigate the effectiveness of the developed biosensor, the blood sorbitol level of type II diabetic rats prepared by streptozotocin administration was measured with the developed sensor. The values of sorbitol were highly correlated with the values measured by the F-kit of food analysis and that we confirmed the sorbitol concentration could be quantified using the developed biosensor. Furthermore, the aldose reductase inhibitor "eparlrestat", which is a therapeutic drug that suppresses the accumulation of sorbitol, was administered to diabetic rats, and the blood sorbitol level was measured with the developed biosensor. As a result, the blood glucose level was high in both the treated group and the non-treated group, but the blood sorbitol level in the treated group decreased. The results suggest that the measurement of the sorbitol level with the developed biosensor in addition to the blood glucose level enables evaluation of complications like diabetic neuropathy. In the future, we expected that the developed sorbitol biosensor will be miniaturized, the pretreatment method for blood samples will be simplified, and it will be applied to the development of therapeutic agents for diabetic complications and personalized medicine.

Enzyme-embedded electrospun fiber sensor of hydrophilic polymer for fluorometric ethanol gas imaging in vapor phase.

Non-invasive measurement of volatile organic compounds (VOCs) emitted from living organisms is a powerful technique for diagnosing health conditions or diseases in humans. Bio-based gas sensors are suitable for the sensitive and selective measurement of a target VOC from a complex mixture of VOCs. Conventional bio-based sensors are normally prepared as wet-type probes to maintain proteins such as enzymes in a stable state, resulting in limitations in the commercialization of sensors, their operating environment, and performance. In this study, we present an enzyme-based fluorometric electrospun fiber sensor (eFES) mesh as a gas-phase biosensor in dry form. The eFES mesh targeting ethanol was fabricated by simple one-step electrospinning of polyvinyl alcohol with an alcohol dehydrogenase and an oxidized form of nicotinamide adenine dinucleotide. The enzyme embedded in the eFES mesh worked actively in a dry state without pretreatment. Substrate specificity was also maintained, and the sensor responded well to ethanol with a sufficient dynamic range. Adjustment of the pH and coenzyme quantity in the eFES mesh also affected enzyme activity. The dry-form biosensor-eFES mesh-will open a new direction for gas-phase biosensors because of its remarkable performance and simple fabrication, which is advantageous for commercialization.

Effect of weight-bearing in bicruciate-retaining total knee arthroplasty during high-flexion activities.

BACKGROUND: To evaluate the effect of weight-bearing on the kinematics of the bicruciate-retaining total knee arthroplasty design during high knee flexion activities. METHODS: The kinematics of 21 bicruciate-retaining total knee arthroplasties were evaluated under fluoroscopy, with two- and three-dimensional image registrations, during squatting (weight-bearing) and active-assisted knee flexion (non-weight-bearing). The following variables were measured: knee range of motion, axis of femoral rotation and varus-valgus angle relative to the tibial component, anteroposterior translation of the medial and lateral contact points, and the kinematic pathway of the joint surfaces. FINDINGS: From 20° to 100° of flexion, the femoral external rotation during weight-bearing was larger than that during non-weight-bearing. There were no differences in the varus-valgus angles between the two conditions. From 10° to 50° of flexion, the medial contact point during weight-bearing was located posterior to the point of contact during non-weight-bearing; this difference between the two weight-bearing conditions was significant. From 0° to 90° of flexion, the lateral contact point in weight-bearing was located posterior to the contact point in non-weight-bearing; this difference between the two weight-bearing conditions was also significant. INTERPRETATION: The anteroposterior position of the medial and lateral contact points of the bicruciate-retaining total knee arthroplasty design was significantly more posterior in the mid-range of knee flexion in weight-bearing than in non-weight-bearing. However, no anterior translation of the bicruciate-retaining total knee arthroplasty design was observed. Therefore, bicruciate-retaining total knee arthroplasty appears to have good anteroposterior stability throughout the range of knee flexion, regardless of the weight-bearing condition.

ReFeaFi: Genome-wide prediction of regulatory elements driving transcription initiation.

Regulatory elements control gene expression through transcription initiation (promoters) and by enhancing transcription at distant regions (enhancers). Accurate identification of regulatory elements is fundamental for annotating genomes and understanding gene expression patterns. While there are many attempts to develop computational promoter and enhancer identification methods, reliable tools to analyze long genomic sequences are still lacking. Prediction methods often perform poorly on the genome-wide scale because the number of negatives is much higher than that in the training sets. To address this issue, we propose a dynamic negative set updating scheme with a two-model approach, using one model for scanning the genome and the other one for testing candidate positions. The developed method achieves good genome-level performance and maintains robust performance when applied to other vertebrate species, without re-training. Moreover, the unannotated predicted regulatory regions made on the human genome are enriched for disease-associated variants, suggesting them to be potentially true regulatory elements rather than false positives. We validated high scoring "false positive" predictions using reporter assay and all tested candidates were successfully validated, demonstrating the ability of our method to discover novel human regulatory regions.

Ascending spinal tract formation in chick embryo originating from different spinal regions.

The present study aimed to assess spinal tract formation in neurons originating from cervical (C7), brachial (C14), and thoracic (T4) regions, with the lumbar (LS2) region as a reference, in a chick embryo. For the assessment of the spinal tracts, we introduced a vector expressing human placental alkaline phosphatase into progenitor cells generated after neural tube closure and belonging to the above segments, using in ovo electroporation. The ascending axons took primarily similar paths: dorsal commissural, ventral commissural, and dorsal non-commissural paths, with some variance depending on their originating segments. Some populations of non-commissural neurons later extended their axons following a ventral path. The elongation rates of these axons are primarily constant and tended to increase over time; however, some variations depending on the originating segments were also observed. Some of the dorsally ascending axons entered into the developing cerebellum, and spinocerebellar neurons originating from T4 projected their axons into the cortex of the cerebellum differently from those from LS2. These results unveil an overall picture of early ascending spinal tract formation.

Biochemical Methanol Gas Sensor (MeOH Bio-Sniffer) for Non-Invasive Assessment of Intestinal Flora from Breath Methanol.

Methanol (MeOH) in exhaled breath has potential for non-invasive assessment of intestinal flora. In this study, we have developed a biochemical gas sensor (bio-sniffer) for MeOH in the gas phase using fluorometry and a cascade reaction with two enzymes, alcohol oxidase (AOD) and formaldehyde dehydrogenase (FALDH). In the cascade reaction, oxidation of MeOH was initially catalyzed by AOD to produce formaldehyde, and then this formaldehyde was successively oxidized via FALDH catalysis together with reduction of oxidized form of ß-nicotinamide adenine dinucleotide (NAD+). As a result of the cascade reaction, reduced form of NAD (NADH) was produced, and MeOH vapor was measured by detecting autofluorescence of NADH. In the development of the MeOH bio-sniffer, three conditions were optimized: selecting a suitable FALDH for better discrimination of MeOH from ethanol in the cascade reaction; buffer pH that maximizes the cascade reaction; and materials and methods to prevent leaking of NAD+ solution from an AOD-FALDH membrane. The dynamic range of the constructed MeOH bio-sniffer was 0.32-20 ppm, which encompassed the MeOH concentration in exhaled breath of healthy people. The measurement of exhaled breath of a healthy subject showed a similar sensorgram to the standard MeOH vapor. These results suggest that the MeOH bio-sniffer exploiting the cascade reaction will become a powerful tool for the non-invasive intestinal flora testing.