Super-resolved 3D Tracking of Cargo Transport Through Nuclear Pore Complexes by Astigmatism Imaging

This protocol describes a two-color astigmatic imaging approach that enables direct 3D visualization of cargo transport trajectories relative to a super-resolved octagonal double-ring scaffold structure of the nuclear pore complex (NPC). Though astigmatism imaging is commonly achieved via a cylindrical lens, this protocol utilizes an adaptive optics (AO) system, which enables optimization of the astigmatism for the precision needs of the experiment as well as correction of the focal mismatch arising from chromatic aberrations in multi-color applications. With this approach, single particle spatial precision values in x, y, and z are typically 5-20 nm, and these depend on astigmatism, photon level and position in z. The method enables resolution of transport conduits through the ~60 nm diameter pore of NPCs by particle tracking on the millisecond timescale. The success of this approach is enabled by the high rigidity of fully active NPCs within the nuclear envelope of permeabilized cells. For a detailed application of this protocol, please refer to https://www.nature.com/articles/s41556-021-00815-6. The figure and table numbers in this protocol that are indicated with an “NCB” prefix (e.g., NCB Figure X) refer to the figures and table in this reference paper.

Aerial Grasping and Transport Using an Unmanned Aircraft (UA) Equipped with an Industrial Suction Gripper

Duetotheavailabilityof highly efficient unmanned aircraft (UA) and the advancement of the necessary technologies, the use of UA for object manipulation and cargo transport is becoming a more and more relevant research area. A reliable identification and localization of cargo and interaction objects as well as maintaining the required flight precision are essential to guarantee a successful object handling. Within this paper we demonstrate the successful application of an autonomous UA equipped with a lightweight suction gripper for object interaction. We discuss the approach used for precise localization as well as the identification and pose estimation of individual gripping objects. Concluding, the overall system performance is evaluated within an industrial-oriented use case.

Extreme value analysis of intracellular cargo transport by motor proteins

In the long axon of a neuron, cargo transport between the cell body and terminal synaptic region are mainly supported by the motor proteins kinesin and dynein, which are nano-sized drivers. Synaptic materials packed as cargos are anterogradely transported to the synaptic region by kinesin, whereas materials accumulated at the axon terminals are returned to the cell body by dynein. Extreme value analysis, typically used for disaster prevention in our society, was applied to analyze the velocity of kinesin and dynein nanosized drivers to disclose their physical properties in living cells.

Evaluation of port workers’ general awareness of dangerous cargo transport

The International Maritime Dangerous Goods (IMDG) Code training is mandatory for all shore-side personnel involved in the handling and transport of dangerous cargoes by sea. This study aims to measure and evaluate the port workers’ dangerous cargo transport general awareness level. For this purpose, the “Dangerous Cargo Transport General Awareness Questionnaire” was developed and applied to trained 100 port workers face-to-face. The questionnaire was proven to be valid and reliable by content validity, construct validity, internal consistency reliability, and test-retest reliability analyses. Four factors were determined as a result of Explanatory Factor Analysis. Discriminant analyses were performed using the Student’s t-test and One Way ANOVA test. Pearson correlation analysis was carried out to determine the relationship between factors. Simple linear regression analysis was used for modeling the relationship between factors. SPSS 24.0 was utilized to conduct the analyses. According to analysis results, port workers’ dangerous cargo transport general awareness does not differ depending on age, position, and education. There is a weak positive or a moderate positive correlation between factors. Port workers have an average and above-average level of general awareness. It is highlighted that increasing this level is possible by increasing the frequency of dangerous cargo transport training. Suggestions are offered for more effective training.

Unconventional COPII-mediated secretory protein trafficking continues in the absence of the Sar1 GTPase

Coat protein complex II (COPII) plays an integral role in the packaging of secretory cargoes within membrane-enclosed transport carriers that leave the endoplasmic reticulum (ER) from discrete membrane subdomains. Lipid bilayer remodeling necessary for this process is driven initially by membrane penetration of the coat subunit Sar1 and further stabilized by assembly of a multi-layer complex of several COPII proteins. However, the relative contributions of these distinct factors to transport carrier formation and protein trafficking remain unclear. Here, we demonstrate that anterograde cargo transport from the ER continues in the absence of Sar1, although the unconventional carriers that form fail to efficiently deliver their contents to subsequent compartments in the secretory pathway. Instead, cargoes accumulate immediately adjacent to the perinuclear Golgi under these conditions, together with components of the COPII coat. Our findings highlight new mechanisms by which COPII promotes transport carrier biogenesis and strongly suggests that the Sar1 GTPase plays a critical role in transport carrier uncoating ahead of membrane fusion and secretory cargo delivery at acceptor compartments.