Application of DNA technology to regulate the transcription of disease-related genes has important therapeutic potential. The transcription factor NFkB plays a pivotal role in the transactivation of inflammatory and adhesion molecule genes, leading to vascular lesion formation. Double-stranded DNA with high affinity for NFkB may be introduced as 'decoy' cis elements to bind NFkB and block the activation of genes mediating inflammation, resulting in effective drugs for treating intimal hyperplasia. In this study, we tested the feasibility of NFkB decoy therapy to treat neointimal formation in a porcine coronary artery balloon injury model as a pre-clinical study.An angioplasty catheter was introduced into the left anterior descending coronary artery of the pig to cause vascular injury. First, we tested the feasibility of transfection of FITC-labeled NFkB decoy ODN using a hydrogel balloon catheter. Fluorescence due to NFkB decoy ODN could be detected throughout the medial layer. Therefore, we transfected NFkB decoy ODN into the balloon-injured LAD using a hydrogel catheter. Histological evaluation demonstrated that the neointimal area in the balloon-injured artery was significantly reduced by NFkB decoy ODN as compared to scrambled decoy ODN at 1 week after single transfection, accompanied by a significant reduction in PCNA-positive stained cells (Po0.01). Interestingly, the reduction of ICAMpositive staining was observed, accompanied by the inhibition of migration of macrophages. Of importance, intravascular ultrasound (IVUS) confirmed that neointimal area in the balloon-injured artery was significantly reduced by NFkB decoy ODN at 4 weeks after transfection (Po0.01). Interestingly, the inhibition of neointimal area was only limited to the lesion transfected with NFkB decoy ODN, while other lesions without NFkB decoy ODN demonstrated a marked increase in neointimal formation.Here, we report the successful in vivo transfer of NFkB decoy ODN using a hydrogel catheter to inhibit vascular lesion formation in balloon-injured porcine coronary artery.

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The anatomical design of the human foot is considered to facilitate generation of bipedal walking. However, how the morphology and structure of the human foot actually contribute to generation of bipedal walking remains unclear. In the present study, we investigated the three-dimensional kinematics of the foot bones under a weight-bearing condition using cadaver specimens, to characterize the innate mobility of the human foot inherently prescribed in its morphology and structure. Five cadaver feet were axially loaded up to 588 N (60 kgf), and radiographic images were captured using a biplane X-ray fluoroscopy system. The present study demonstrated that the talus is medioinferiorly translated and internally rotated as the calcaneus is everted owing to axial loading, causing internal rotation of the tibia and flattening of the medial longitudinal arch in the foot. Furthermore, as the talus is internally rotated, the talar head moves medially with respect to the navicular, inducing external rotation of the navicular and metatarsals. Under axial loading, the cuboid is everted simultaneously with the calcaneus owing to the osseous locking mechanism in the calcaneocuboid joint. Such detailed descriptions about the innate mobility of the human foot will contribute to clarifying functional adaptation and pathogenic mechanisms of the human foot.

BackgroundQuantifying detailed 3-dimensional (3D) kinematics of the foot in contact with the ground during locomotion is crucial for understanding the biomechanical functions of the complex musculoskeletal structure of the foot. Biplanar X-ray fluoroscopic systems and model-based registration techniques have recently been employed to capture and visualise 3D foot bone movements in vivo, but such techniques have generally been performed manually. In the present study, we developed an automatic model-registration method with biplanar fluoroscopy for accurate measurement of 3D movements of the skeletal foot.MethodsThree-dimensional surface models of foot bones were generated prior to motion measurement based on computed tomography. The bone models generated were then registered to biplanar fluoroscopic images in a frame-by-frame manner using an optimisation technique, to maximise similarity measures between occluding contours of the bone surface models with edge-enhanced fluoroscopic images, while avoiding mutual penetration of bones. A template-matching method was also introduced to estimate the amount of bone translation and rotation prior to automatic registration.ResultsWe analysed 3D skeletal movements of a cadaver foot mobilized by a robotic gait simulator. The 3D kinematics of the calcaneus, talus, navicular and cuboid in the stance phase of the gait were successfully reconstructed and quantified using the proposed model-registration method. The accuracy of bone registration was evaluated as 0.27 ± 0.19 mm and 0.24 ± 0.19° (mean ± standard deviation) in translation and rotation, respectively, under static conditions, and 0.36 ± 0.19 mm and 0.42 ± 0.30° in translation and rotation, respectively, under dynamic conditions.ConclusionsThe measurement was confirmed to be sufficiently accurate for actual analysis of foot kinematics. The proposed method may serve as an effective tool for understanding the biomechanical function of the human foot during locomotion.

Dishevelled (DVL) is a central factor in the Wnt signaling pathway, which is highly conserved among various organisms. DVL plays important roles in transcriptional activation in the nucleus, but the molecular mechanisms underlying their nuclear localization remain unclear. In the present study, we identified IQGAP1 as a regulator of DVL function. In Xenopus embryos, depletion of IQGAP1 reduced Wnt-induced nuclear accumulation of DVL, and expression of Wnt target genes during early embryogenesis. The domains in DVL and IQGAP1 that mediated their interaction are also required for their nuclear localization. Endogenous expression of Wnt target genes was reduced by depletion of IQGAP1 during early embryogenesis, but notably not by depletion of other IQGAP family genes. Moreover, expression of Wnt target genes caused by depletion of endogenous IQGAP1 could be rescued by expression of wild-type IQGAP1, but not IQGAP1 deleting DVL binding region. These results provide the first evidence that IQGAP1 functions as a modulator in the canonical Wnt signaling pathway.

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Abstractβ-Catenin is an important component of the Wnt signalling pathway. As dysregulation or mutation of this pathway causes many diseases, including cancer, the β-Catenin level is carefully regulated by the destruction complex in the Wnt signalling pathway. However, the mechanisms underlying the regulation of β-Catenin ubiquitination and degradation remain unclear. Here, we find that WNK (With No Lysine [K]) kinase is a potential regulator of the Wnt signalling pathway. We show that WNK protects the interaction between β-Catenin and the Glucose-Induced degradation Deficient (GID) complex, which includes an E3 ubiquitin ligase targeting β-Catenin, and that WNK regulates the β-Catenin level. Furthermore, we show that WNK inhibitors induced β-Catenin degradation and that one of these inhibitors suppressed xenograft tumour development in mice. These results suggest that WNK is a previously unrecognized regulator of β-Catenin and a therapeutic target of cancer.

The past ten years have seen the rapid expansion of the field of biohybrid robotics. By combining engineered, synthetic components with living biological materials, new robotics solutions have been developed that harness the adaptability of living muscles, the sensitivity of living sensory cells, and even the computational abilities of living neurons. Biohybrid robotics has taken the popular and scientific media by storm with advances in the field, moving biohybrid robotics out of science fiction and into real science and engineering. So how did we get here, and where should the field of biohybrid robotics go next? In this perspective, we first provide the historical context of crucial subareas of biohybrid robotics by reviewing the past 10+ years of advances in microorganism-bots and sperm-bots, cyborgs, and tissue-based robots. We then present critical challenges facing the field and provide our perspectives on the vital future steps towards creating autonomous living machines.