Cutting off ciliary protein import:intraflagellar transport after dendritic femtosecond-laser ablation

Primary cilia, organelles protruding from the surface of eukaryotic cells, act as cellular antennae to detect and transmit signals from the extracellular environment. They are built and maintained by continuous cycles of intraflagellar transport (IFT), where ciliary proteins are transported between the ciliary base and tip. These proteins originate from the cell body because cilia lack protein synthesis machinery. How input from the cell body affects IFT and ciliary function is not well understood. Here, we use femtosecond-laser ablation to perturb the dendritic input of proteins to chemosensory cilia in living Caenorhabditis elegans. Using fluorescence microscopy, we visualize and quantify the real-time response of ciliary proteins to dendritic ablation. We find that the response occurs in three distinct stages. First, IFT dynein is activated within seconds, redistributing IFT components toward the ciliary base; second, the ciliary axoneme shortens and motors slow down; and third, motors leave the cilium. Depletion of ATP by adding azide also results in IFT slowdown and IFT components leaving the cilium, but not in activation of retrograde IFT. These results indicate that laser ablation triggers a specific mechanism important for IFT regulation that allows the cilium to rapidly adapt to changes in the outside environment

Development of an Algorithm for Determining Defects in Cast-in-Place Piles Based on the Data Analysis of Low Strain Integrity Testing

Low strain integrity testing for pile quality control, based on the analysis of elastic waves, is one of the most common methods, due to its high efficiency. However, it also has a number of limitations that should be taken into account during pile testing. For additional study of the method and its effectiveness, an experimental site was constructed, consisting of ten cast-in-place piles with embedded defects. When analyzing field data, pile defects were not identified. For further analysis of the problem, as well as for interpreting the results and identifying pile defects, a cluster analysis method, the so-called ANN-classifier, is proposed. This paper describes the results of creating an algorithm for the recognition of defects and their localization in cast-in-place piles. It is proposed that use of the characteristic points of the spectrum of the signal as the input vector of the ANN classifier, and the type of pile defect as the output vector, is optimal. The results of the study led to the conclusion that the ANN-classifier can be used as the main tool for automatic interpretation of the results obtained by low strain integrity testing

Development of an Algorithm for Determining Defects in Cast-in-Place Piles Based on the Data Analysis of Low Strain Integrity Testing

Low strain integrity testing for pile quality control, based on the analysis of elastic waves, is one of the most common methods, due to its high efficiency. However, it also has a number of limitations that should be taken into account during pile testing. For additional study of the method and its effectiveness, an experimental site was constructed, consisting of ten cast-in-place piles with embedded defects. When analyzing field data, pile defects were not identified. For further analysis of the problem, as well as for interpreting the results and identifying pile defects, a cluster analysis method, the so-called ANN-classifier, is proposed. This paper describes the results of creating an algorithm for the recognition of defects and their localization in cast-in-place piles. It is proposed that use of the characteristic points of the spectrum of the signal as the input vector of the ANN classifier, and the type of pile defect as the output vector, is optimal. The results of the study led to the conclusion that the ANN-classifier can be used as the main tool for automatic interpretation of the results obtained by low strain integrity testing

Wavelet Analysis for Evaluating the Length of Precast Spliced Piles Using Low Strain Integrity Testing

The difficulties with the application of low strain integrity testing for evaluating the length of driven precast piles of two sections justify the need for new data acquisition and analysis techniques. The standard time domain analysis of the recorded signals may not allow for distinguishing the desired responses from a pile toe and a splice. In this paper, we propose the use of a set of hammers of different weights and tip materials that will provide various sensitivities of the test to a pile splice. To further analyze the collected data, we study the distributions of phase angles obtained using complex continuous wavelet transform. The characteristic phase shifts that distribute from higher to lower frequencies can be interpreted as responses from a pile toe and a splice. To verify the proposed approaches, a series of numerical simulations were performed using the finite element method for the driven pile models with the different properties of a splice zone. Numerical simulation results show that the pile splices are clearly identified when using the shorter input pulses which can be generated by light hammers with a hard head material. The total length of a simulated pile with a 1 mm air gap between sections was undetectable by standard data analysis approaches and was evaluated when analyzing the wavelet phase angle distributions. Numerically validated data acquisition and analysis techniques were applied to field data analysis and allowed us to confidently identify the length of two-section piles grouped with a pile cap

Wavelet Analysis for Evaluating the Length of Precast Spliced Piles Using Low Strain Integrity Testing

The difficulties with the application of low strain integrity testing for evaluating the length of driven precast piles of two sections justify the need for new data acquisition and analysis techniques. The standard time domain analysis of the recorded signals may not allow for distinguishing the desired responses from a pile toe and a splice. In this paper, we propose the use of a set of hammers of different weights and tip materials that will provide various sensitivities of the test to a pile splice. To further analyze the collected data, we study the distributions of phase angles obtained using complex continuous wavelet transform. The characteristic phase shifts that distribute from higher to lower frequencies can be interpreted as responses from a pile toe and a splice. To verify the proposed approaches, a series of numerical simulations were performed using the finite element method for the driven pile models with the different properties of a splice zone. Numerical simulation results show that the pile splices are clearly identified when using the shorter input pulses which can be generated by light hammers with a hard head material. The total length of a simulated pile with a 1 mm air gap between sections was undetectable by standard data analysis approaches and was evaluated when analyzing the wavelet phase angle distributions. Numerically validated data acquisition and analysis techniques were applied to field data analysis and allowed us to confidently identify the length of two-section piles grouped with a pile cap

Replication Data for: IFT cargo and motors associate sequentially with IFT trains to enter cilia

The dataset contains the data and the MATLAB scripts associated with the manuscript titled IFT cargo and motors associate sequentially with IFT trains to enter cilia Here we provide scripts and data corresponding to four different aspects: (1) single-molecule tracking data for all the IFT-components studied in the paper (2) numerical simulations of single-molecule entry events to estimate real pause time. (3) numerical simuations for obtaining 1D projection information from simulated 2D localization data along a hollow cylinder (4) Spline fitting to transform global xy-coordinates of single-molecule tracks to spline coordinates The data and codes are orgainized in separate folders. Each folder contains a ReadMe.txt file which details the data (and structure) and the accompanying script(s). Data is stored as .csv (or xlsx) files and scripts are written in MATLAB (The Math Works, Inc., R2021a)

Single-Molecule Fluorescence Microscopy in Sensory Cilia of Living Caenorhabditis elegans

Intracellular transport of organelles and biomolecules is vital for several cellular processes. Single-molecule fluorescence microscopy can illuminate molecular aspects of the dynamics of individual biomolecules that remain unresolved in ensemble experiments. For example, studying single-molecule trajectories of moving biomolecules can reveal motility properties such as velocity, diffusivity, location and duration of pauses, etc. We use single-molecule imaging to study the dynamics of microtubule-based motor proteins and their cargo in the primary cilia of living C. elegans. To this end, we employ standard fluorescent proteins, an epi-illuminated, widefield fluorescence microscope, and primarily open-source software. This chapter describes the setup we use, the preparation of samples, a protocol for single-molecule imaging in primary cilia of C. elegans, and data analysis.</p

Replication Data for:A small excitation window allows long-duration single-molecule imaging, with reduced background autofluorescence, in C. elegans neurons

The dataset contains the data and the MATLAB scripts associated with publication titled A small excitation window allows long-duration single-molecule imaging, with reduced background autofluorescence, in C. elegans neurons. Here we provide scripts and data corresponding to three different aspects: (1) the characterization of the technique, small-window illumination microscopy (SWIM), that was developed in this work (2) the numerical simulations to explain how SWIM reduces the amount of out-of-focus autofluoresence background during imaging. (3) the statistical analysis of the motion of vesicles moving back-and-forth in a C. elegans dendrite, imaged using SWIM. The data and codes are orgainized in separate folders, with the titles indicating the figure the data belongs to. In addition, there are a couple of folders containing essential scripts used for analysis of single-molecule tracks. Each folder contains a ReadMe.txt file which details the data (and structure) and the accompanying script(s). Data is stored as .csv (or xlsx) files and scripts are written in MATLAB (The Math Works, Inc., R2021a)

Marketing Research of Personnel Motivation and Pharmacy Chains

The purpose of the study is to analyze the motivation of pharmacy and pharmacy chain employees to identify problem areas and develop instructions to improve the business management system. As a result of the study, motivation was found to be one of the main conditions for the effective work of a pharmaceutical organization, while the most important components of the employee motivation system are positive external factors such as financial motivation. Based on the results of the study, recommendations have been developed to improve the construction of the motivation system.El propósito del estudio es analizar la motivación de los empleados de farmacias y cadenas de farmacias para identificar áreas problemáticas y desarrollar instrucciones mejorando el sistema de gestión empresarial. Como resultado del estudio, se descubrió que la motivación es una de las principales condiciones para el trabajo eficaz de una organización farmacéutica, mientras que los componentes más importantes del sistema de motivación para los empleados son factores externos positivos como la Motivación financiera. Con base en los resultados del estudio, se han desarrollado recomendaciones para mejorar la construcción del sistema de motivación