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Abstract Wastewater surveillance is an effective tool for monitoring community spread of COVID-19 and other diseases. Quantitative PCR (qPCR) analysis for wastewater surveillance is more susceptible to mutations in target genome regions than binary PCR analysis for clinical surveillance. The SARS-CoV-2 concentrations in wastewater estimated by N1 and N2 qPCR assays started to diverge around July 2022 in data from different sampling sites, analytical methods, and analytical laboratories in Japan. On the basis of clinical genomic surveillance data and experimental data, we demonstrate that the divergence is due to two mutations in the N1 probe region, which can cause underestimation of viral concentrations. We further show that this inaccuracy can be alleviated if the qPCR data are analyzed with the second derivative method or the Cy0 method instead of the crossing point method.

This paper examines the machining accuracy when simultaneously machining pilot holes and threads, and the machining accuracy when machining threads after pilot hole drilling. To verify them, cutting forces were measured in the tool rotation coordinate system using a wireless holder. To examine the influence of the machining area of the tool bottom edge machining the pilot hole, three patterns of pilot hole diameters (φ0 mm, 7.0 mm, and 8.5 mm) were used. The results showed that when the pilot hole diameter is φ0 mm, the cutting force in the plane direction does not increase significantly compared to the case with a pilot hole due to the offset effect of the cutting forces generated by the facing cutting edges when machining the bottom edge. Therefore, if chip entrapment can be prevented, it is highly possible to omit the pilot hole, improve machining efficiency, and maintain machining accuracy. It was also found that tool vibration could be reduced. However, when the pilot hole diameter is Φ0, the load in the tool axis direction increases. Therefore, it was also found that machine rigidity and tool life must be considered due to the increased load during machining. These results are summarized in this paper.

Background and aims: Protein C is a plasma protein, and its active form regulates blood coagulation. The recommended unit of protein C activity is IU/mL; however, some laboratories use percentage. Some deficiencies cannot be detected owing to measurement principles. This study sought to quantify protein C activity levels and overcome the limitations of the current measurements. Materials and methods: Our protein C activity measurement method mimicked the blood coagulation cascade and used a thrombin-specific chromogenic reagent. The control was prepared by adding protein C to the protein C deficient plasma. The calibration curve was plotted as the increase in the absorbance per minute and the concentration of protein C in the control. Statistical tests were performed to compare our method with the current chromogenic method. Results: A calibration curve was constructed (y = −0.0132x + 0.14, R2 = 0.9987, n = 10). The statistical results of our method suggested non-inferiority when compared to the current chromogenic method (α = 0.05). Conclusion: The quantitative measurement was performed using plasma samples. Our method provides the possibility of expressing protein C activity quantitatively and detecting deficiencies that cannot be detected using the current chromogenic method.

In recent years, the development of technologies for "connected factories" based on the Internet of Things (IoT) has become important, and industrial robots are required to operate autonomously based on sensing technologies and their data. In this paper, we focused on the polishing work with a cooperative robot, and discussed a method to integrate the technology of the developed wireless vibration monitoring system with the internal information of the servo. In particular, we attempted to construct a novel finishing system that utilizes the internal information of the force sensor installed in the base of the cooperative robot not only to detect human contact but also to improve the accuracy of machining. We estimated the rough-endmilling state from the in-process monitoring the vibration with a wireless tool holder system and the internal information of the servo obtained during endmilling operation, and also integrated the finishing process using the newly developed system considering rough endmilling information with a cooperative robot.

Cos lettuce was grown under different spectral photon flux density distribution (SPFD) change patterns with blue- and/or red light-emitting diode (LED) irradiation with a 24-hour cycle. Twelve treatments were designed with a combination of four relative SPFD (RSPFD) change patterns and three photosynthetic photon flux density (PPFD) levels. The RSPFD change patterns were as follows: BR/BR, simultaneous blue- and red-light irradiation (BR) for 24 h; R/BR, red-light monochromatic irradiation (R) for 12 h followed by 12 hours of BR; B/BR, blue-light monochromatic irradiation (B) for 12 hours followed by 12 hours of BR; and B/R, 12 hours of B followed by 12 hours of R. Each RSPFD change pattern was conducted at three daily average photosynthetic photon flux densities (PPFDave) of 50, 100, and 200 µmol·m−2·s−1. The RSPFD change patterns that included B (B/BR and B/R) resulted in elongated leaves. A low ratio of active phytochrome to total phytochrome under B was considered the reason for leaf elongation. Shoot dry weight was significantly greater under the RSPFD change patterns that included B when the PPFDave was 200 µmol·m−2·s−1. The leaf elongation caused by B would have increased the amount of light received and thereby promoted growth. However, excessive leaf elongation caused the plants to fall, and growth was not promoted under the RSPFD change patterns that included B when the PPFDave was 50 µmol·m−2·s−1. Thus, 12-hour B promoted growth under conditions in which leaf elongation leads to increases in the amount of light received.

Leaves acclimate to day-to-day fluctuating levels of photosynthetic photon flux density (PPFD) by adjusting their morphological and physiological parameters. Accurate estimation of these parameters under day-to-day fluctuating PPFD conditions benefits crop growth modeling and light environment management in greenhouses, although it remains challenging. We quantified the relationships between day-to-day PPFD changes over 6 days and light acclimation parameters for cucumber seedling leaves, including leaf mass per area (LMA), chlorophyll (Chl) a/b ratio, maximum net photosynthetic rate (Pnmax), maximum rate of ribulose-1,5-bisphosphate (RuBP) carboxylase/oxygenase (Vcmax), and maximum rate of electron transport (Jmax). The last two parameters reflect the capacity of the photosynthetic partial reactions. We built linear regression models of these parameters based on average or time-weighted averages of daily PPFDs. For time-weighted averages of daily PPFDs, the influence of daily PPFD was given a specific weight. We employed three types of functions to calculate this weight, including linear, quadratic, and sigmoid derivative types. We then determined the trend of weights that estimated each parameter most accurately. Moreover, we introduced saturating functions to calibrate the average or time-weighted averages of daily PPFDs, considering that light acclimation parameters are usually saturated under high PPFDs. We found that time-weighted average PPFDs, in which recent PPFD levels had larger weights than earlier levels, better estimated LMA than average PPFDs. This suggests that recent PPFDs contribute more to LMA than earlier PPFDs. Except for the Chl a/b ratio, the average PPFDs estimated Pnmax, Vcmax, and Jmax with acceptable accuracy. In contrast, time-weighted averages of daily PPFDs did not improve the estimation accuracy of these four parameters, possibly due to their low response rates and plasticity. Calibrating functions generally improved estimation of Chl a/b ratio, Vcmax, and Jmax because of their saturating tendencies under high PPFDs. Our findings provide a reasonable approach to quantifying the extent to which the leaves acclimate to day-to-day fluctuating PPFDs, especially the extent of LMA.

The morphology of plants growing under combined blue- and red-light irradiation is affected by the presence or absence of time slots of blue- and red-light mono-irradiation. The purposes of this study were to investigate the morphology and growth of cos lettuce grown under light irradiation combining several durations of blue and red light simultaneously and independent mono-irradiations of blue and red light during the day, and to clarify the effects of the durations of blue-light mono-irradiation and blue-light irradiation. Young cos lettuce seedlings were grown under 24-h blue-light irradiation with a photosynthetic photon flux density (PPFD) of 110μmol m−2 s−1 (B+0R) or under 24-h blue-light irradiation with a PPFD of 100μmol m−2 s−1 supplemented with 8 (B+8R), 16 (B+16R), and 24-h (B+24R) red-light irradiation with PPFDs of 30, 15, and 10μmol m−2 s−1, respectively (Experiment 1). The daily light integral was 9.50mol m−2 in all treatments. In Experiment 1, leaf elongation was promoted as the duration of red-light irradiation decreased and the duration of blue-light mono-irradiation increased. The maximum shoot dry weight was observed under the B+8R treatment. Growth was likely promoted by the expansion of the light-receptive area caused by moderate leaf elongation without tilting. In Experiment 2, young cos lettuce seedlings were grown as for Experiment 1, but blue- and red-light irradiation intensities were reversed (R+0B, R+8B, R+16B, and R+24B). Leaf elongation was promoted by the absence of blue-light irradiation (R+0B). The leaf surface was increasingly flattened, and the shoot dry weight was enhanced, as the duration of blue-light irradiation increased. Thus, cos lettuce leaf morphology may be manipulated by adjusting each duration of blue-light mono-irradiation, red-light mono-irradiation, and blue- and red-light simultaneous irradiation, which can, in turn, promote cos lettuce growth.

The regulation and coordination of F-actin networks are not well understood. Here, the authors show an antagonistic relationship between endosomal F-actin assembly and cortical bundle integrity during epithelial tube maturation that is regulated by kinase and phosphatase activity modifying WASH.

The effects of photosynthetic photon flux density (PPFD) fluctuations in sunlight have already been investigated; however, the spectral photon flux density distribution (SPD) has hardly been considered. Here, sunlight SPD fluctuations recorded for 200 min in October in Tokyo, Japan were artificially reproduced using an LED-artificial sunlight source system. The net photosynthetic rate (Pn) of cucumber leaves under reproduced sunlight was measured and compared with the Pn estimated from a steady-state PPFD–Pn curve for the same leaves. The measured and estimated Pn agreed except when the PPFD was low, where the measured Pn was lower than the estimated Pn. The ratio of measured Pn to estimated Pn was 0.94–0.95 for PPFD ranges of 300–700 μmol m–2 s–1, while the value was 0.98–0.99 for 900–1,300 μmol m–2 s–1, and the overall ratio was 0.97. This 3% reduction in the measured Pn compared with the Pn estimated from a steady-state PPFD–Pn curve was significantly smaller than the approximately 20–30% reduction reported in previous experimental and simulation studies. This result suggests that the loss of integral net photosynthetic gain under fluctuating sunlight can vary among days with different fluctuation patterns or may be non-significant when fluctuations in both PPFD and relative SPD of sunlight are taken into consideration.

To drive LEDs for plant cultivation using an alternating-current (AC) power supply, full-wave rectification (FWR) is a reasonable method to supply a unidirectional forward current to LEDs because of the simple configuration and low energy loss of the rectification circuit. We grew cos lettuce hydroponically using a white LED light source that emitted continuous light, 100 Hz square-wave (SW) pulsed light, or pulsed light generated with FWR of 50 Hz sine-wave AC with the same averaged photosynthetic photon flux density of 150 µmol m-2 s-1. The results showed that shoot fresh weight, shoot dry weight, leaf area, and number of leaves did not differ significantly among the treatments. Plants grown under FWR pulsed light showed similar net photosynthetic rates under continuous light and SW pulsed light. Shoot fresh weight per power consumption was estimated to be significantly greater with FWR pulsed light than with continuous light, and we concluded that the use of FWR pulsed light without elaborated transformation to a flat waveform direct current is a promising lighting method to reduce the lighting cost. [ABSTRACT FROM AUTHOR]

A custom-made fluorescence measurement system was constructed for non-destructive and quantitative estimation of green fluorescent protein (GFP) transiently expressing in a Nicotiana benthamiana leaf with a viral vector. The system comprised a digital camera, a blue LED lamp, and a long wavelength-pass filter to acquire the brightness intensity of the green channel (G value) of a leaf surface image as a measure of fluorescence intensity. The genes encoding GFP and tobamoviral vector were transferred to N. benthamiana leaves 42 days post seeding by syringe infiltration with Agrobacterium for transient expression. The destructively measured GFP content in leaves as the ground-truth data started to increase at 3-4 days post infiltration (DPI), once showed a plateau at 6-8 DPI, and then increased again until 11 DPI. Regression analysis demonstrated that the r² of the linear regression between the ground-truth GFP content and the G value was 0.86-0.87 when the G value at a given DPI was calibrated with the initial leaf color at 0 DPI for the same leaf. These results indicate that the constructed system and method could potentially be used to quantitatively and non-destructively estimate the amount of GFP transiently expressing in intact N. benthamiana leaves. [ABSTRACT FROM AUTHOR]

Nowadays, a monitoring technology has attracted attention in the factory automation fields regarding IoT (Internet of Things). However, it is difficult to monitor the process information from a round tool during rotating operation in machine tools. We therefore develop a novel tool holder equipped with a wireless communication function to monitor tool temperature and vibration. In the present report, we attempt to measure the inner temperature in drill tool and investigate the influence of feed rate and cutting speed on it. Moreover, we attempt to measure the tool vibrations in the rotational and radial direction in countersinking process. As a result, we demonstrated that the developed method with a wireless system is effective to estimate the tool temperature in drilling processes and the tool vibration in countersinking processes.

In this study, gas exchange characteristics and temperature of Nicotiana benthamiana leaves transiently overexpressing hemagglutinin (HA), an influenza vaccine antigen, with an Agrobacterium tumefaciens-assisted viral vector were investigated. Inoculation of leaves with an empty viral vector not containing the HA gene decreased the net photosynthetic rate (Pn) and transpiration rate (T) from 2 to 3 days post-infiltration (DPI) in the A. tumefaciens suspension. Expression of HA with the vector decreased Pn and T to much lower levels until 4 DPI. Such significant decreases were not observed in leaves infiltrated with suspension of A. tumefaciens not carrying the viral vector or in uninfiltrated leaves. Thus, viral vector inoculation itself decreased Pn and T to a certain extent and the HA expression further decreased them. The decreases in Pn and T in empty vector-inoculated and HA expression vector-inoculated leaves were associated with decreases in stomatal conductance, suggesting that the reduction of gas exchange rates was caused at least in part by stomatal closure. More detailed gas exchange and chlorophyll fluorescence analyses revealed that in HA vector-inoculated leaves, the capacity of ribulose-1,5-bisphosphate carboxylase/oxygenase to assimilate CO2 and the capacity of photosynthetic electron transport in planta were downregulated, which contributed also to the decrease in Pn. Leaf temperature (LT) increased in viral vector-inoculated leaves, which was associated with the decrease in T. When HA vector-inoculated leaves were grown at air temperatures (ATs) of 21, 23, and 26°C post-infiltration, HA accumulated earlier in leaves and the days required for HA content to attain its peak became shorter, as AT was higher. The highest LT was found 1–2 days earlier than the highest leaf HA content under all post-infiltration AT conditions. This phenomenon could be applicable in a non-destructive technique to detect the optimum harvesting date for individual plants to determine the day when leaf HA content reaches its maximum level, irrespective of spatiotemporal variation of AT, in a plant growth facility.

􀉹We examined whether the method that we have recently proposed for quantitative estimation of leaf mass per area 􀀉LMA􀀊 in leaves grown under day-to-day changing photosynthetic photon flux density 􀀉PPFD􀀊 could be applied to conditions where PPFD also fluctuated during the daytime. The method utilizes the time-weighted average PPFD 􀀉Qwl􀀊 in which a greater weight was assigned to PPFD levels in recent days than in past days. Cucumber seedlings were treated for six days with seven light treatment patterns of day-to-day PPFD changes, each combined with two diurnal PPFD conditions: constant- and fluctuating-PPFD light during a 16-h d-1 photoperiod. The LMA tended to increase with increasing Qwl and was greater under diurnal constant-PPFD light than fluctuating-PPFD light in each of the seven day-to-day light treatment patterns. Regression analysis revealed a significant and highly linear relationship between LMA and Qwl in each of constant- and fluctuating-PPFD light conditions. Our result demonstrates that the concept of Qwl for quantitative estimation of LMA can be applied to light conditions where the PPFD level fluctuates not only day to day but also during the daytime, although some adjustments would be required to incorporate the effect of daytime PPFD fluctuations properly. [ABSTRACT FROM AUTHOR]

Cucumber seedling growth under constant and fluctuating photosynthetic photon flux density 􀀉PPFD􀀊 light 􀀉CPL and FPL, respectively􀀊 was examined to discuss whether plants can acclimate to CPL versus FPL. The FPL was created in a growth chamber by artificially reproducing fluctuations in ground-level sunlight PPFD using phosphor-converted white LEDs. The daily mean PPFD at the canopy surface and photoperiod were the same for the CPL and FPL. The plants were grown for two weeks, with the former and latter half periods under either CPL or FPL, respectively; there were two treatments for the first week and four with the 2 􀂺 2 combinations for the last week. According to the growth analysis for the first week, the net assimilation rate 􀀉NAR􀀊 tended to be lower under FPL than under CPL, while the leaf area ratio and specific leaf area 􀀉SLA􀀊 were greater under FPL than under CPL. The lower NAR may be due to a delayed response of photosynthesis to a PPFD increase, i.e., photosynthetic induction, and the non-linearity of the PPFD-response curve of photosynthetic rate. The greater SLA was consistent with previous reports, although the mechanism underlying the response was unclear. For the last week, there was no significant difference in those parameters among treatments, possibly because of a change in canopy structure accompanying self and mutual leaf shading. The relative growth rate and shoot dry mass were also not significantly different among treatments. Thus, although cucumber seedlings initially showed physiological and morphological responses to CPL versus FPL distinctively, the responses did not improve growth under continued respective light conditions and may not necessarily be acclimative at the whole-plant growth level. [ABSTRACT FROM AUTHOR]

Developmental potentials of cells are tightly controlled at multiple levels. The embryonic Drosophila airway tree is roughly subdivided into two types of cells with distinct developmental potentials: a proximally located group of multipotent adult precursor cells (P-fate) and a distally located population of more differentiated cells (D-fate). We show that the GATA-family transcription factor (TF) Grain promotes the P-fate and the POU-homeobox TF Ventral veinless (Vvl/Drifter/U-turned) stimulates the D-fate. Hedgehog and receptor tyrosine kinase (RTK) signaling cooperate with Vvl to drive the D-fate at the expense of the P-fate while negative regulators of either of these signaling pathways ensure P-fate specification. Local concentrations of Decapentaplegic/BMP, Wingless/Wnt, and Hedgehog signals differentially regulate the expression of D-factors and P-factors to transform an equipotent primordial field into a concentric pattern of radially different morphogenetic potentials, which gradually gives rise to the distal-proximal organization of distinct cell types in the mature airway.

Nowadays, infrared thermographic technology has been attracting attention in various industrial fields. We therefore focus on it as a novel method for monitoring surface tool temperature to improve end-milling conditions for difficult-to-cut materials. However, a problem has emerged; it is difficult to measure the tool surface temperature when there is a coolant because the coolant prevents monitoring of the surface of the end-mill tool. Thus, we developed a wireless tool holder system equipped with a thermocouple in the end mill to monitor the tool internal temperature under coolant conditions. In the present report, we compared the temperature measured by infrared thermographic imagery with that measured by a wireless tool holder system when end milling the stainless steel under dry coolant conditions. The thermocouple, which has a diameter of 1.0 mm, was used to ensure stable measurement in the proposed wireless tool holder. We obtained the tool temperatures by infrared thermographic imagery and by wireless tool holder equipped with a thermocouple at a sampling time of 1/30 of a second. We confirmed that the tool internal temperature measured by the wireless tool holder agrees with one calculated by FEM model based on infrared thermographic imagery. As a result, we demonstrated that the developed method with a wireless system is effective to estimate the tool temperature in end-milling processes and makes it feasible to measure it under coolant conditions.

The tubular networks of the Drosophila respiratory system and our vasculature show distinct branching patterns and tube shapes in different body regions. These local variations are crucial for organ function and organismal fitness. Organotypic patterns and tube geometries in branched networks are typically controlled by variations of extrinsic signaling but the impact of intrinsic factors on branch patterns and shapes is not well explored. Here, we show that the intersection of extrinsic hedgehog(hh) and WNT/wingless (wg) signaling with the tube-intrinsic Hox code of distinct segments specifies the tube pattern and shape of the Drosophila airways. In the cephalic part of the airways, hh signaling induces expression of the transcription factor (TF) knirps (kni) in the anterior dorsal trunk (DTa1). kni represses the expression of another TF spalt major (salm), making DTa1 a narrow and long tube. In DTa branches of more posterior metameres, Bithorax Complex (BX-C) Hox genes autonomously divert hh signaling from inducing kni, thereby allowing DTa branches to develop as salm-dependent thick and short tubes. Moreover, the differential expression of BX-C genes is partly responsible for the anterior-to-posterior gradual increase of the DT tube diameter through regulating the expression level of Salm, a transcriptional target of WNT/wg signaling. Thus, our results highlight how tube intrinsic differential competence can diversify tube morphology without changing availabilities of extrinsic factors.