Knock-in mouse models have contributed tremendously to our understanding of human disorders. However, generation of knock-in animals requires a significant investment of time and effort. We addressed this problem by developing a novel knock-in system that circumvents several traditional challenges by establishing stem cells with acceptor elements enveloping a particular genomic target. Once established, these acceptor embryonic stem (ES) cells are efficient at directionally incorporating mutated target DNA using modified Cre/lox technology. This is advantageous, because knock-ins are not restricted to one a priori selected variation. Rather, it is possible to generate several mutant animal lines harboring desired alterations in the targeted area. Acceptor ES cell generation is the rate-limiting step, lasting approximately 2 months. Subsequent manipulations toward animal production require an additional 8 weeks, but this delimits the full period from conception of the genetic alteration to its animal incorporation. We call this system a “kick-in” to emphasize its unique characteristics of speed and convenience. To demonstrate the functionality of the kick-in methodology, we generated two mouse lines with separate mutant versions of the voltage-dependent potassium channel Kv7.2 (Kcnq2): p.Tyr284Cys (Y284C) and p.Ala306Thr (A306T); both variations have been associated with benign familial neonatal epilepsy. Adult mice homozygous for Y284C, heretofore unexamined in animals, presented with spontaneous seizures, whereas A306T homozygotes died early. Heterozygous mice of both lines showed increased sensitivity to pentylenetetrazole, possibly due to a reduction in M-current in CA1 hippocampal pyramidal neurons. Our observations for the A306T animals match those obtained with traditional knock-in technology, demonstrating that the kick-in system can readily generate mice bearing various mutations, making it a suitable feeder technology toward streamlined phenotyping.

It is important for construction companies to sustain a productive workforce without sacrificing its health and safety. This study aims to develop a practical judgement method to estimate the workload risk of individual construction workers. Based on studies, we developed a workload model comprising a hygrothermal environment, behavioral information, and the physical characteristics of workers). The construction workers’ heart rate and physical activity were measured using the data collected from a wearable device equipped with a biosensor and an acceleration sensor. This study is the first report to use worker physical activity, age, and the wet bulb globe temperature (WBGT) to determine a worker’s physical workload. The accuracy of this health risk judgment result was 89.2%, indicating that it is possible to easily judge the health risk of workers even in an environment where it is difficult to measure the subject in advance. The proposed model and its findings can aid in monitoring the health impacts of working conditions during construction activities, and thereby contribute toward determining workers’ health damage. However, the sampled construction workers are 12 workers, further studies in other working conditions are required to accumulate more evidence and assure the accuracy of the models.

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Background A construction method has emerged in which a camera is installed around a construction machine, and the operator remotely controls the machine while synchronizing the vibration of the machine with the images seen from the operator's seat using virtual reality (VR) technology. Indices related to changes in heart rate (HR) and physical vibration, such as heart rate variability (HRV) and multiscale entropy (MSE), can then be measured among the operators. As these indices are quantitative measures of autonomic regulation in the cardiovascular system, they can provide a useful means of assessing operational stress. Objective In this study, we aimed to evaluate changes in HR and body vibration of machine operators and investigate appropriate methods of machine operation while considering the psychological load. Methods We enrolled 9 remote operators (18-50 years old) in the experiment, which involved 42 measurements. A construction machine was driven on a test course simulating a construction site, and three patterns of operation—riding operation, remote operation using monitor images, and VR operation combining monitor images and machine vibration—were compared. The heartbeat, body vibration, and driving time of the participants were measured using sensing wear made of a woven film-like conductive material and a three-axis acceleration measurement device (WHS-2). We used HRV analysis in the time and frequency domains, MSE analysis as a measure of the complexity of heart rate changes, and the ISO (International Standards Organization) 2631 vibration index. Multiple regression analysis was conducted to model the relationship among the low frequency (LF)/high frequency (HF) HRV, MSE, vibration index, and driving time of construction equipment. Efficiency in driving time was investigated with a focus on stress reduction. Results Multiple comparisons conducted via the Bonferroni test and Kruskal-Wallis test showed statistically significant differences (P=.05) in HRV-LF/HF, the vibration index, weighted acceleration, motion sickness dose value (MSDVz), and the driving time among the three operation patterns. The riding operation was found to reduce the driving time of the machine, but the operation stress was the highest in this case; operation based on the monitor image was found to have the lowest operation stress but the longest operation time. Multiple regression analysis showed that the explanatory variables (LH/HF), RR interval, and vibration index (MSDVz by vertical oscillation at 0.5-5 Hz) had a negative effect on the driving time (adjusted coefficient of determination R2=0.449). Conclusions A new method was developed to calculate the appropriate operating time by considering operational stress and suppressing the physical vibration within an acceptable range. By focusing on the relationship between psychological load and physical vibration, which has not been explored in previous studies, the relationship of these variables with the driving time of construction machines was clarified.

Objectives: Patients identified as asymptomatic for unilateral spatial neglect (USN) based on paper-and-pen tests nonetheless often collide with objects to their left while walking. This study aimed to investigate chronic USN in subjects who experienced collisions while walking. Methods: Two patients with chronic USN who experienced collisions while walking were evaluated using the Behavioral Inattention Test-conventional (BIT-c). Additionally, the modified Posner task (MPT) was used to evaluate the left and right reaction times. MPT targets randomly appeared either on the side indicated by the cue (valid condition) or on the opposite side (invalid condition). This study used an alternating treatments single-case design. The valid and invalid conditions of the MPT alternated rapidly and randomly to determine differences in reaction time. Statistical analysis compared left and right reaction times using a one-tailed randomization test to study valid and invalid conditions. Results: The total BIT-c score was in the normal range for both subjects, whereas MPT reaction times were higher on the left side than on the right side for the invalid condition. However, for the valid condition, only Case B had increased reaction times on the left side. Conclusions: The MPT valid condition evaluates voluntary attention, whereas the invalid condition evaluates the reorientation of attention. Consequently, for Case A, a left reorientation of attention deficit was observed, whereas, for Case B, left voluntary attention and left reorientation of attention deficits were observed. The MPT results revealed the characteristics of covert neglect signs. USN evaluation would benefit from additional research using MPT.