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Abstract Flowering plant sexual reproduction relies on the communication between the pollen tube and synergid cells to induce pollen tube bursting. During this process, the MILDEW RESISTANCE LOCUS-O (MLO) protein NORTIA (NTA) is polarly trafficked from the Golgi, where it is inactive, to the filiform apparatus, where it is functional in synergids. MLOs were recently described as calcium channels and have been proposed to be negatively regulated through calmodulin (CaM) binding at a conserved C-terminal calmodulin-binding domain (CaMBD). To determine whether CaM binding is necessary for MLO function during pollen tube reception, C-terminal truncations and CaMBD point mutations were made in NTA. Point mutations were also generated in a constitutively filiform apparatus-localized chimeric NTA containing the MLO1 C-terminus. In this study, we demonstrate that mutating the MLO1 and NTA CaMBD reduces the ability for MLOs to function during pollen tube reception. This is in part due to altered subcellular localization of the CaMBD mutants in synergids. We showed that the CaMBD is not necessary for Golgi localization of MLOs, but is necessary for efficient trafficking and total protein accumulation at the filiform apparatus. Our results suggest an additional role for CaM binding as a regulator of MLO trafficking in addition to its previously proposed role as a negative regulator of MLO Ca2+ channel activity. 

Amputees’ preferences for prosthesis settings are critical not only for their psychological well-being but also for long-term adherence to device adoption and health. Although active lower-limb prostheses can provide enhanced functionality than passive devices, little is known about the mechanism of preferences for settings in active devices. Therefore, a think-aloud study was conducted on three amputees to unravel their preferences for a powered robotic knee prosthesis during user-guided auto-tuning. The inductive thematic analysis revealed that amputee patients were more likely to use their own passive device rather than the intact leg as the reference for the natural walking that they were looking for in the powered device. There were large individual differences in factors influencing naturalness. The mental optimization of preference decisions was mostly based on the noticeableness of the differences between knee profiles. The implications on future design and research in active prostheses were discussed. 

Prostheses help amputees to maintain physical health and quality of life. Prosthesis wearers’ satisfaction and adherence to the prosthesis are closely related to the preferences for prosthesis tuning settings. However, the underlying factors that contribute to the preferences were under-explored. In this study, two able-bodied participants were asked to change the robotic prosthesis settings to their preferred state and the think-aloud technique with a mixed-method approach was used to reveal the contributing factors of preferences. We found that physical perception (e.g., positions of the prosthetic foot, balance, and stability) and subjective feelings (e.g., comfortableness, satisfaction, confidence, and worries) were two major factors. Experiences with the intact leg and other profiles were used as anchors for their preference levels. Preferences may also differ with situational context such as walking speed. The saturation points were reached with no strong approach motivation. The implications for prosthesis design and research were discussed. 

Agrobacterium transfers T-DNA to plants where it may integrate into the genome. Non-homologous end-joining (NHEJ) has been invoked as the mechanism of T-DNA integration, but the role of various NHEJ proteins remains controversial. Genetic evidence for the role of NHEJ in T-DNA integration has yielded conflicting results. We propose to investigate the formation of T-circles as a proxy for understanding T-DNA integration. T-circles are circular double-strand T-DNA molecules, joined at their left (LB) and right (RB) border regions, formed in plants. We characterized LB-RB junction regions from hundreds of T-circles formed in Nicotiana benthamiana or Arabidopsis thaliana . These junctions resembled T-DNA/plant DNA junctions found in integrated T-DNA: Among complex T-circles composed of multiple T-DNA molecules, RB-RB/LB-LB junctions predominated over RB-LB junctions; deletions at the LB were more frequent and extensive than those at the RB; microhomology was frequently used at junction sites; and filler DNA, from the plant genome or various Agrobacterium replicons, was often present between the borders. Ku80 was not required for efficient T-circle formation, and a VirD2 ω mutation affected T-circle formation and T-DNA integration similarly. We suggest that investigating the formation of T-circles may serve as a surrogate for understanding T-DNA integration. 

Abstract California experienced a historic run of nine consecutive landfalling atmospheric rivers (ARs) in three weeks’ time during winter 2022/23. Following three years of drought from 2020 to 2022, intense landfalling ARs across California in December 2022–January 2023 were responsible for bringing reservoirs back to historical averages and producing damaging floods and debris flows. In recent years, the Center for Western Weather and Water Extremes and collaborating institutions have developed and routinely provided to end users peer-reviewed experimental seasonal (1–6 month lead time) and subseasonal (2–6 week lead time) prediction tools for western U.S. ARs, circulation regimes, and precipitation. Here, we evaluate the performance of experimental seasonal precipitation forecasts for winter 2022/23, along with experimental subseasonal AR activity and circulation forecasts during the December 2022 regime shift from dry conditions to persistent troughing and record AR-driven wetness over the western United States. Experimental seasonal precipitation forecasts were too dry across Southern California (likely due to their overreliance on La Niña), and the observed above-normal precipitation across Northern and Central California was underpredicted. However, experimental subseasonal forecasts skillfully captured the regime shift from dry to wet conditions in late December 2022 at 2–3 week lead time. During this time, an active MJO shift from phases 4 and 5 to 6 and 7 occurred, which historically tilts the odds toward increased AR activity over California. New experimental seasonal and subseasonal synthesis forecast products, designed to aggregate information across institutions and methods, are introduced in the context of this historic winter to provide situational awareness guidance to western U.S. water managers. 

Land surface albedo is a significant regulator of climate. Changes in land use worldwide have greatly reshaped landscapes in the recent decades. Deforestation, agricultural development, and urban expansion alter land surface albedo, each with unique influences on shortwave radiative forcing and global warming impact (GWI). Here, we characterize the changes in landscape albedo-induced GWI (GWIΔα) at multiple temporal scales, with a special focus on the seasonal and monthly GWIΔα over a 19-year period for different land cover types in five ecoregions within a watershed in the upper Midwest USA. The results show that land cover changes from the original forest exhibited a net cooling effect, with contributions of annual GWIΔα varying by cover type and ecoregion. Seasonal and monthly variations of the GWIΔα showed unique trends over the 19-year period and contributed differently to the total GWIΔα. Cropland contributed most to cooling the local climate, with seasonal and monthly offsets of 18% and 83%, respectively, of the annual greenhouse gas emissions of maize fields in the same area. Urban areas exhibited both cooling and warming effects. Cropland and urban areas showed significantly different seasonal GWIΔα at some ecoregions. The landscape composition of the five ecoregions could cause different net landscape GWIΔα. 

Introduction Tuning of lower-limb (LL) robotic prosthesis control is necessary to provide personalised assistance to each human wearer during walking. Prostheses wearers’ adaptation processes are subjective and the efficiency largely depends on one’s mental processes. Therefore, beyond physical motor performance, prosthesis personalisation should consider the wearer’s preference and cognitive performance during walking. As a first step, it is necessary to examine the current measures of cognitive performance when a wearer walks with an LL prosthesis, identify the gaps and methodological considerations, and explore additional measures in a walking setting. In this protocol, we outlined a scoping review that will systematically summarise and evaluate the measures of cognitive performance during walking with and without LL prosthesis. Methods and analysis The review process will be guided and documented by CADIMA, an open-access online data management portal for evidence synthesis. Keyword searches will be conducted in seven databases (Web of Science, MEDLINE, BIOSIS, SciELO Citation Index, ProQuest, CINAHL and PsycINFO) up to 2020 supplemented with grey literature searches. Retrieved records will be screened by at least two independent reviewers on the title-and-abstract level and then the full-text level. Selected studies will be evaluated for reporting bias. Data on sample characteristics, type of cognitive function, characteristics of cognitive measures, task prioritisation, experimental design and walking setting will be extracted. Ethics and dissemination This scoping review will evaluate the measures used in previously published studies thus does not require ethical approval. The results will contribute to the advancement of prosthesis tuning processes by reviewing the application status of cognitive measures during walking with and without prosthesis and laying the foundation for developing needed measures for cognitive assessment during walking. The results will be disseminated through conferences and journals.