Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)1.

In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field

Investigation of the Effect of Initial Graft Tension During Anterior Talofibular Ligament Reconstruction on Ankle Kinematics, Laxity, and In-situ Force

Category: Basic Sciences/Biologics Introduction/Purpose: Ankle sprains are the most common sports injuries, and anterior talofibular ligament (ATFL) injury comprised 85% of all ankle sprains. Most patients recover with conservative treatment, but 20% of them progress to chronic ankle instability. Some studies have reported that anatomic reconstruction using a tendon graft is one of the best procedures to restore the ankle to its condition before symptom development. However, the effect of initial graft tension during ATFL reconstruction is still unclear. Therefore, the objective of this study was to investigate the effect of the initial graft tension during ATFL reconstruction. Methods: Eight fresh-frozen cadaveric ankle specimens were subjected to passive plantarflexion (PF)-dorsiflexion (DF) movement from 15° DF to 30° PF using the 6-degree-freedom robotic system. In addition, 60 N of anterior-posterior load, 1.7 Nm of inversion-eversion (IV-EV) torque, and 1.7 Nm of internal-external rotation (IR-ER) torque were applied to the ankle. During testing, 3-dimensional paths of the ankle were recorded simultaneously. Furthermore, in-situ forces of the ATFL and reconstructed graft were calculated using the principle of superposition. A repeated experiment was designed with the intact condition (intact), ATFL transection, and ATFL reconstruction with four different initial graft tensions (10 N, 30 N, 50 N, and 70 N). Results: AP laxity, IV-EV laxity and IR-ER laxity with ATFL transection was significantly greater than those with intact. In ATFL transection, the talus was significantly translated anteriorly with inversion and internal rotations under passive PF-DF motion compared with intact. Kinematic patterns and laxity in ATFL reconstruction with initial tension of 10 N and 30 N almost imitated intact, but in ATFL reconstruction with initial tension 70 N, the talus was significantly translated with external rotation compared with intact. As the initial graft tension during ATFL reconstruction increased, in-situ force of the reconstructed graft tended to increase during PF-DF motion. In-situ force of the reconstructed graft tension was significantly greater with initial tensions of 50 N, and 70 N than with intact during PF-DF motion (Figure 1). Conclusion: ATFL deficiency altered ankle kinematics and laxity. Although the optimal initial graft tension during ATFL reconstruction might restore ankle kinematics and laxity, excessive initial graft tension caused abnormal kinematics and laxity. Furthermore, the reconstructed graft tension increased as the initial tension increased. Initial tension during ATFL reconstruction has the important effect of imitating the normal ankle condition. We suggest that over-tensioning during ATFL reconstruction should be avoided in order to imitate the conditions of a normal ankle

Subject-Specific Orientations of the Talocrural Joint Axes Estimated from the Morphology of the Talar Trochlea

Category: Ankle Introduction/Purpose: The individual morphology variations in the talar trochlea influence subject-specific kinematics of the talocrural joint. In particular, it is suggested that the medial to lateral radius ratios of the anterior and posterior talar trochlea determine the orientations of the dorsiflexion and plantarflexion axes of the talocrural joint, respectively. However, the characteristics of the orientations of these axes of the talocrural joint have not been elucidated. The aims of this study were to assess the variations in the medial to lateral radius ratios of the anterior and posterior talar trochlea and to estimate the subject- specific orientations of the dorsiflexion and plantarflexion rotational axes of the talocrural joint. Methods: Computed tomography (CT) images of forty-nine unilateral adult human male tali were enrolled in this study. The three-dimensional bone models of the talus were reconstructed from the CT images. Four circles were fitted to medial and lateral edges of each anterior and posterior trochlea, respectively. Then, the radii of curvatures of these circles were measured. The medial to lateral radius ratios of the anterior and posterior trochlea were also calculated, respectively. In addition, the line connecting the centers of the medial and lateral circles fitted to anterior trochlea was defined as a dorsiflexion axis of the talocrural joint, and that of posterior trochlea was also defined as a plantarflexion counterpart. The coronal and transverse angles of these axes were calculated. Results: At the anterior trochlea, the medial to lateral radius ratio was ranged from 0.43 to 0.92. This result of the calculated values meant that the all of the lateral curvature of radius was larger than medial counterpart. On the other hand, at the posterior trochlea, the medial to lateral radius ratio was ranged from 0.58 to 1.63. The coronal and transverse angles of dorsiflexion axis were on average 19.6 ± 7.3° (range, -3.9° to 38.9°) and 10.3 ± 4.6° (range, -0.9° to 23.2°), respectively. The coronal and transverse angles of plantarflexion axis were on average -1.5 ± 12.8° (range, -31.8° to 31.0°) and 7.5 ± 4.5° (range, -3.4° to 17.4°). Conclusion: This study found different features between orientations of the dorsiflexion and plantarflexion axes of the talocrural joint. Concretely, in all the talus specimens, the dorsiflexion axis of the talocrural joint is inclined downwards and laterally on the coronal plane, and inclined backwards and laterally on the transverse plane. On the other hand, there were two types of the plantarflexion axis, one was a type whose axis inclined as the same direction as dorsiflexion axis, and the other was a type whose axis inclined downwards and medially on the coronal plane and inclined backwards and laterally on the transverse plane

Factors Related to the Occurrence of Osteochondral Lesions of the Talus by 3-dimensional Bone Morphology of the Ankle

Category: Ankle Introduction/Purpose: Repeated microtrauma is thought to play a major role in the occurrence of osteochondral lesions of the talus (OLTs), but much remains unknown. Two-dimensional assessments of the relationship between ankle bone morphology and OLTs are occasionally seen. The purpose of this study was to evaluate the bone morphology of the ankle in OLT 3-dimensionally using three-dimensional computed tomography (3DCT), and to investigate the factors related to the occurrence of OLTs. Methods: The subjects were 19 patients (19 ankles) who underwent surgery for medial OLTs (OLT group). They included 13 men and 6 women. A healthy group without ankle disease served as a control group with the same number of 19 ankles. Three-dimensional ankle joint models were made based on DICOM data obtained with CT images. In the 3D model, the medial malleolus articular surface and the tibial plafond surface, the medial surface of the trochlea of the talus, and the lateral surface of the trochlea of the talus were defined. The tibial axis-medial malleolus (TMM) angle, the medial malleolus surface area (MMA), the medial malleolus volume (MMV), and the anterior opening angle of the talus were measured 3-dimensionally and compared in the OLT and control groups. Results: The mean TMM angle was significantly larger in the OLT group (34.2 ± 4.4°) than in the control group (29.2 ± 4.8°; p = 0.002). The mean MMA was significantly smaller in the OLT group (219.8 ± 42.4 mm2) than in the control group (280.5 ± 38.2 mm2; p < 0.001). The mean MMV was significantly smaller in the OLT group (2119.9 ± 562.5 mm3) than in the control group (2646.4 ± 631.4 mm3; p = 0.01). The mean anterior opening angle of the talus was significantly larger in the OLT group (15.4 ± 3.9°) than in the control group (10.2 ± 3.6°; p < 0.001). Conclusion: It was shown with 3DCT measurements that, in medial OLT patients, the medial malleolus opens distally, the MMA and MMV are small, and the talus anterior opening angle was significantly larger than in controls. This study suggests the possibility that the 3D bone morphology of both the mortise and tenon of the ankle joint are closely related to the occurrence of OLTs

Pulmonary lymphoepithelial cyst with no prior HIV infection: A case report

Most lymphoepithelial cysts (LECs) occur in the salivary glands and are considered one of the autoimmune syndromes caused by the human immunodeficiency virus (HIV). In this report, we present a case of pulmonary LEC without prior HIV infection, paying special attention to radiographic features. A chest radiograph revealed an oval mass with a smooth surface, localized in the left lower lung field, which was in direct contact with the diaphragm. Computed tomography showed an oval homogenous mass with a smooth surface in the lower left lobe. Further, magnetic resonance imaging demonstrated that the mass was a homogeneous internal structure with a smooth surface and a slightly high signal in T2-weighted images and a slightly low signal in T1-weighted images. Surgical resection was performed, and pathological examination confirmed the diagnosis of a pulmonary LEC. To the best of our knowledge, no cases of pulmonary LECs without prior HIV infection have been reported in the literature to date, therefore, the case presented here is considered very rare and informative

Automatic switching between the AAI and the DDD algorithm can prevent repetitive non-reentrant ventriculoatrial synchrony

A 67-year-old man with non-obstructive hypertrophic cardiomyopathy had received an implantable cardioverter-defibrillator (ICD) for an unstable, sustained ventricular tachycardia (VT) induced by programmed stimulation during an electrophysiological study 5 years earlier. An intracardiac electrogram recorded by the ICD revealed repetitive, non-reentrant ventriculoatrial synchrony (RNRVAS) associated with hypotension. Electrophysiologic and hemodynamic studies indicated that RNRVAS was induced and reproducibly termed by a single ventricular extrastimulus from the right ventricular apex. Following attainment of the elective replacement indicator, we replaced the ICD with another having managed ventricular pacing, which automatically switched AAI and DDD, thereby avoiding unnecessary ventricular pacing. Thus far, the patient has not experienced further RNRVAS. Thus, we believe that automatic switching between AAI and DDD can prevent RNRVAS