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The Lrp and MarR families are two groups of transcriptional regulators widely distributed among prokaryotes. However, the hierarchical-regulatory relationship between the Lrp family and the MarR family remains unknown. Our previous study found that an Lrp (SACE_Lrp) from Saccharopolyspora erythraea indirectly repressed the biosynthesis of erythromycin. In this study, we characterized a novel MarR family protein (SACE_6745) from S. erythraea, which is controlled by SACE_Lrp and plays a direct regulatory role in erythromycin biosynthesis and export. SACE_Lrp directly regulated the expression of marR by specifically binding a precise site OM (5'-CTCCGGGAACCATT-3'). Gene disruption of marR increased the production of erythromycin by 45% in S. erythraea A226. We found that MarR has direct DNA-binding activity for the promoter regions of the erythromycin biosynthetic genes, as well as an ABC exporter SACE_2701-2702 which was genetically proved to be responsible for erythromycin efflux. Disruption of SACE_Lrp in industrial S. erythraea WB was an efficient strategy to enhance erythromycin production. Herein, we jointly engineered SACE_Lrp and its target MarR by deleting marR in WBΔSACE_Lrp, resulting in 20% increase in erythromycin yield in mutant WBΔLrpΔmarR compared to WBΔSACE_Lrp, and 39% to WB. Overall, our findings provide new insights into the hierarchical-regulatory relationship of Lrp and MarR proteins and new avenues for coordinating antibiotic biosynthesis and export by joint engineering regulators in actinomycetes. Key points: • The hierarchical-regulatory relationship between SACE_Lrp and MarR was identified. • MarR directly controlled the expression of erythromycin biosynthesis and export genes. • Joint engineering of SACE_Lrp-MarR regulatory element enhanced erythromycin production. [ABSTRACT FROM AUTHOR]

Victims of urban violence are at risk of developing Posttraumatic Stress Disorder (PTSD), one of the most debilitating consequences of violence. Considering that PTSD may be associated with inefficient selection of defensive responses, it is important to understand the relation between motor processing and PTSD. The present study aims to investigate the extent to which the severity of posttraumatic stress symptoms (PTSS) is related to motor preparation against visual threat cues in victims of urban violence. Participants performed a choice reaction time task while ignoring a picture that could be threating or neutral. The EEG indices extracted were the motor-related amplitude asymmetry (MRAA) in the alpha frequency range, and the lateralized readiness potential (LRP). We observed a linear relation between longer LRP latency and a slower reaction time, selectively during threat processing (compared to neutral) in low PTSS, but not in high PTSS participants. Alpha MRAA suppression and the PTSS were also linearly related: the smaller the alpha MRAA suppression in the threat condition relative to neutral, the greater the PTSS. These results provide evidence that threatening cues affect motor processing that is modulated by the severity of PTSS in victims of urban violence.

A critical issue in executive control is how the nervous system exerts flexibility to inhibit a prepotent response and adapt to sudden changes in the environment. In this study, force measurement was used to capture “partial” unsuccessful trials that are highly relevant in extending the current understanding of motor inhibition processing. Moreover, a modified version of the stop-signal task was used to control and eliminate potential attentional capture effects from the motor inhibition index. The results illustrate that the non-canceled force and force rate increased as a function of stop-signal delay (SSD), offering new objective indices for gauging the dynamic inhibitory process. Motor response (time and force) was a function of delay in the presentation of novel/infrequent stimuli. A larger lateralized readiness potential (LRP) amplitude in go and novel stimuli indicated an influence of the novel stimuli on central motor processing. Moreover, an early N1 component reflects an index of motor inhibition in addition to the N2 component reported in previous studies. Source analysis revealed that the activation of N2 originated from inhibitory control associated areas: the right inferior frontal gyrus (rIFG), pre-motor cortex, and primary motor cortex. Regarding partial responses, LRP and error-related negativity (ERNs) were associated with error correction processes, whereas the N2 component may indicate the functional overlap between inhibition and error correction. In sum, the present study has developed reliable and objective indices of motor inhibition by introducing force, force-rate and electrophysiological measures, further elucidating our understandings of dynamic motor inhibition and error correction.

Xenorhabdus hominickii ANU1 is known to be an entomopathogenic bacterium symbiotic to nematode Steinernema monticolum. Another bacterial strain X. hominickii DY1 was isolated from a local population of S. monticolum. This bacterial strain X. hominickii DY1 was found to exhibit high insecticidal activities against lepidopteran and coleopteran species after hemocoelic injection. However, these two X. hominickii strains exhibited significant variations in insecticidal activities, with ANU1 strain being more potent than DY1 strain. To clarify their virulence difference, bacterial culture broths of these two strains were compared for secondary metabolite compositions. GC-MS analysis revealed that these two strains had different compositions, including pyrrolopyrazines, piperazines, cyclopeptides, and indoles. Some of these compounds exhibited inhibitory activities against phospholipase A2 to block eicosanoid biosynthesis and induce significant immunosuppression. They also exhibited significant insecticidal activities after oral feeding, with indole derivatives being the most potent. More kinds of indole derivatives were detected in the culture broth of ANU1 strain. To investigate variations in regulation of secondary metabolite production, expression level of leucine-responsive regulatory protein (Lrp), a global transcription factor, was compared. ANU1 strain exhibited significantly lower Lrp expression level than DY1 strain. To assess genetic variations associated with secondary metabolite synthesis, bacterial loci encoding non-ribosomal protein synthase and polyketide synthase (NRPS-PKS) were compared. Three NRPS and four PKS loci were predicted from the genome of X. hominickii. The two bacterial strains exhibited genetic variations (0.12∼0.67%) in amino acid sequences of these NRPS-PKS. Most NRPS-PKS genes exhibited high expression peaks at stationary phase of bacterial growth. However, their expression levels were significantly different between the two strains. These results suggest that differential virulence of the two bacterial strains is caused by the difference in Lrp expression level, leading to difference in the production of indole compounds and other NRPS-PKS-associated secondary metabolites.

A critical issue in executive control is how the nervous system exerts flexibility to inhibit a prepotent response and adapt to sudden changes in the environment. In this study, force measurement was used to capture "partial" unsuccessful trials that are highly relevant in extending the current understanding of motor inhibition processing. Moreover, a modified version of the stop-signal task was used to control and eliminate potential attentional capture effects from the motor inhibition index. The results illustrate that the non-canceled force and force rate increased as a function of stop-signal delay (SSD), offering new objective indices for gauging the dynamic inhibitory process. Motor response (time and force) was a function of delay in the presentation of novel/infrequent stimuli. A larger lateralized readiness potential (LRP) amplitude in go and novel stimuli indicated an influence of the novel stimuli on central motor processing. Moreover, an early N1 component reflects an index of motor inhibition in addition to the N2 component reported in previous studies. Source analysis revealed that the activation of N2 originated from inhibitory control associated areas: the right inferior frontal gyrus (rIFG), pre-motor cortex, and primary motor cortex. Regarding partial responses, LRP and error-related negativity (ERNs) were associated with error correction processes, whereas the N2 component may indicate the functional overlap between inhibition and error correction. In sum, the present study has developed reliable and objective indices of motor inhibition by introducing force, force-rate and electrophysiological measures, further elucidating our understandings of dynamic motor inhibition and error correction. [ABSTRACT FROM AUTHOR]

Not only automation of manufacturing processes but also automation of automation procedures itself become increasingly relevant to automation research. In this context, automated capability assessment, mainly leveraged by deep learning systems driven from 3D CAD data, have been presented. Current assessment systems may be able to assess CAD data with regards to abstract features, e.g. the ability to automatically separate components from bulk goods, or the presence of gripping surfaces. Nevertheless, they suffer from the factor of black box systems, where an assessment can be learned and generated easily, but without any geometrical indicator about the reasons of the system's decision. By utilizing explainable AI (xAI) methods, we attempt to open up the black box. Explainable AI methods have been used in order to assess whether a neural network has successfully learned a given task or to analyze which features of an input might lead to an adversarial attack. These methods aim to derive additional insights into a neural network, by analyzing patterns from a given input and its impact to the network's output. Within the NeuroCAD Project, xAI methods are used to identify geometrical features which are associated with a certain abstract feature. Within this work, a sensitivity analysis (SA), the layer-wise relevance propagation (LRP), the Gradient-weighted Class Activation Mapping (Grad-CAM) method as well as the Local Interpretable Model-Agnostic Explanations (LIME) have been implemented in the NeuroCAD environment, allowing not only to assess CAD models but also to identify features which have been relevant for the network's decision. In the medium run, this might enable to identify regions of interest supporting product designers to optimize their models with regards to assembly processes. [ABSTRACT FROM AUTHOR]

Xenorhabdus hominickii ANU1 is known to be an entomopathogenic bacterium symbiotic to nematode Steinernema monticolum. Another bacterial strain X. hominickii DY1 was isolated from a local population of S. monticolum. This bacterial strain X. hominickii DY1 was found to exhibit high insecticidal activities against lepidopteran and coleopteran species after hemocoelic injection. However, these two X. hominickii strains exhibited significant variations in insecticidal activities, with ANU1 strain being more potent than DY1 strain. To clarify their virulence difference, bacterial culture broths of these two strains were compared for secondary metabolite compositions. GC-MS analysis revealed that these two strains had different compositions, including pyrrolopyrazines, piperazines, cyclopeptides, and indoles. Some of these compounds exhibited inhibitory activities against phospholipase A2 to block eicosanoid biosynthesis and induce significant immunosuppression. They also exhibited significant insecticidal activities after oral feeding, with indole derivatives being the most potent. More kinds of indole derivatives were detected in the culture broth of ANU1 strain. To investigate variations in regulation of secondary metabolite production, expression level of leucine-responsive regulatory protein (Lrp), a global transcription factor, was compared. ANU1 strain exhibited significantly lower Lrp expression level than DY1 strain. To assess genetic variations associated with secondary metabolite synthesis, bacterial loci encoding non-ribosomal protein synthase and polyketide synthase (NRPS-PKS) were compared. Three NRPS and four PKS loci were predicted from the genome of X. hominickii. The two bacterial strains exhibited genetic variations (0.12∼0.67%) in amino acid sequences of these NRPS-PKS. Most NRPS-PKS genes exhibited high expression peaks at stationary phase of bacterial growth. However, their expression levels were significantly different between the two strains. These results suggest that differential virulence of the two bacterial strains is caused by the difference in Lrp expression level, leading to difference in the production of indole compounds and other NRPS-PKS-associated secondary metabolites. [ABSTRACT FROM AUTHOR]

Chuchu Zhao,* Ermei Lu,* Xiaoli Hu, Huihui Cheng, Jian-An Zhang, Xueqiong Zhu Department of Obstetrics and Gynecology, The Second Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325027, China *These authors contributed equally to this work Objective: Our previous research has shown that the expression of S100 calcium-binding protein A9 (S100A9) in tumor cells was associated with neoadjuvant chemotherapy sensitivity in cervical squamous cell carcinoma. In the present study, we altered the expression of S100A9 through infecting lentivirus, investigated its effect on the chemosensitivity to cisplatin of cervical cancer cells and then made a primary exploration of the involved mechanism.Materials and methods: Lentivirus was employed to upregulate and downregulate S100A9 expression in SiHa cells. The protein expression level of apoptotic-related proteins Bcl-2 and Bax, drug resistance-related proteins multiple drug resistance protein 1 (MRP1), P glycoprotein (P-gp), glutathione-S-transferase-π (GST-π), lung resistance-related protein (LRP), and FOXO1 signaling pathway related proteins was detected by Western blot. The CCK-8 assay was used to examine chemosensitivity to cisplatin, and the proportion of apoptosis cells was analyzed by the flow cytometry.Results: S100A9 overexpression could obviously increase the IC50 value of SiHa cells to cisplatin and decrease the apoptosis rate induced by cisplatin. Downregulation of S100A9 led to the opposite results. In S100A9 overexpression SiHa cells, the expression level of Bcl-2, LRP, GST-π, p-AKT, p-ERK, p-FOXO1, and Nanog was significantly increased, while FOXO1 expression was decreased. The opposite results were observed in S100A9 knockdown SiHa cells.Conclusion: Downregulation of S100A9 could significantly increase apoptosis rate, resulting in enhancing sensitivity of SiHa cells to cisplatin, which may be related to Bcl-2, GST-π, and LRP protein and by altering the AKT/ERK-FOXO1-Nanog signaling pathway. Keywords: S100A9, cervical cancer, Bcl-2, GST-π, LRP, FOXO1, cisplatin, chemosensitivity

Recently conducted research demonstrated the potential use of mouse dynamics as a behavioral biometric for user authentication systems. However, the state-of-the-art methods in this field rely on classical machine learning methods that necessitate the design of hand crafted mouse features for feature extraction. To simplify the feature extraction process, we leverage various deep learning architectures for mouse movement sequences classification, including convolutional networks, recurrent networks, and a hybrid model which combines convolutional and recurrent layers. It is known that the training of these networks with random initialization of weights on small datasets will produce models that perform poorly. Therefore, we consider a two-dimensional convolutional neural network that allows transfer learning, which is a domain adaptation technique effective for learning on small datasets. Although employing such architecture may seem counterintuitive, since the temporal information is discarded from the input data, the architecture has outperformed all the other deep architectures investigated, as well as a classical machine learning method. In order to understand the features learned, we adopt the layer-wise relevance propagation (LRP) algorithm to compute relevance scores for each part of the mouse curves. In addition, the models are measured for their usability and effectiveness in realistic scenarios. [ABSTRACT FROM AUTHOR]

Green synthesis of silver chloride nanoparticles (AgCl NPs) using commercial yeast extract has been carried out. The physico–chemical characterizations of AgCl NPs were carried out by UV–visible spectroscopy, XRD, FTIR, HR-SEM equipped with EDAX and TEM. In vitro efficacy of anti-mycobacterial properties of AgCl NPs were determined by agar well diffusion and Luciferase Reporter Phage (LRP) assay were used against Mycobacteriumsmegmatis (MC2155) and Mycobacteriumtuberculosis H37Rv respectively. This shows that the AgCl NPs have potential anti-myobacterial activity against Mycobacterium tuberculosis H37Rv. [ABSTRACT FROM AUTHOR]

Wide application of gonadotropin-releasing hormone (GnRH) agonists and antagonists for clinical purposes determines their effects on ovarian signaling pathways. Our study aimed to determine the localization, expression levels of Wnt signaling members in the pubertal and adult mouse ovary and the impact of GnRH antagonist cetrorelix on these signaling members. 0.5 mg/kg of cetrorelix was injected to 3-and 6-week-old mice for 2 weeks. At the end of injection, ovaries from 5 (5Ce)- to 8-week (8Ce)-old mice were embedded in paraffin for immunohistochemistry and homogenized for western blot to compare with control (5C–8C) and sham groups (5S–8S). WNT2 and WNT4 showed higher expression in thecal and stromal cells in adult mouse ovaries and only WNT4 expression was affected by cetrorelix. FZD1 was localized mainly in oocytes of pubertal ovaries and granulosa cells and oocytes of adult ovaries. FZD1 was reduced by cetrorelix in pubertal ovaries. FZD4 was abundantly localized in thecal and stromal cells of all groups and protein level was not affected by cetrorelix. LRP-6 was expressed mainly in oocytes and stromal cells of pubertal, oocytes of adult ovaries and its expression was reduced by cetrorelix in adult ovaries. CTNNB1 intensity in granulosa cells was the lowest in pubertal and the highest in adult ovaries and its expression was decreased by cetrorelix in adult ovaries. Cetrorelix affected the expression of specific members of the Wnt signaling depending on the developmental stage of mice, pointing out its possible interaction with gonadotropins during pubertal and adult stages. [ABSTRACT FROM AUTHOR]

An innovative optimization strategy by means of hyper-heuristics is proposed. It consists of a parallel combination of three metaheuristics. In view of the need both to escape from local optima and to achieve high diversity, the algorithm cooperatively combines simulated annealing with genetic algorithms and ant colony optimization. A location routing problem (LRP), which aims at the design of transport networks, was adopted for the performance evaluation of the proposed algorithm. Information exchanges took place effectively between the metaheuristics and speeded up the search process. Moreover, the parallel implementation was useful since it allowed several metaheuristics to run simultaneously, thus achieving a significant reduction in the computational time. The algorithmic efficiency and effectiveness were ratified for a medium-sized city. The proposed optimization algorithm not only accelerated computations, but also helped to improve solution quality. [ABSTRACT FROM AUTHOR]

• Experimental study on Non-destructive characterization of cylindrical shell by probing. • Geometric imperfections of cylindrical shells is measured to aid the identification of appropriate probe locations. • Least resistance path and probing forces are used to single out a best probe location. • Peak load carrying capacity of the cylindrical shell under axial compression is predicted using probing procedure. In the present study, the prediction of the critical buckling load of an axially compressed cylindrical shell is investigated based on the novel non-destructive probing method with the aid of Measured Geometric Imperfections (MGI). A laser point scanner is used to obtain the imperfection data. The imperfection data is analyzed to find a sample space of points where the buckling is likely to initiate. The optimum location for probing is determined from the sample set by finding the Least Resistance Path (LRP) to probing using two methods, one based on the Energy barrier and the other from the reaction forces observed corresponding to deep probe advancements. It is found that MGI can aid the procedure by improving the selection of the probing points and hence the prediction accuracy; However, MGI does not eliminate the need for multiple probing. The study has shown the possibility of obtaining multiple probing locations that can predict the peak load within acceptable levels of accuracy. [ABSTRACT FROM AUTHOR]

Post-processing methods that rely on fusion-grided forecast products can reduce systematic biases from Numerical Weather Prediction (NWP) precipitation forecasts. However, these methods also limit the capability to forecast precipitation accurately at local stations. We constructed a Station-based Precipitation Post-processing Model (SPPM) that utilizes deep-learning algorithms, predominantly convolutional layers and ResNet modules. Based on 390 meteorological stations in North China and European Centre for Medium-Range Weather Forecasts Highest-resolution (ECMWF-HRES) forecast data, the SPPM utilizes multi-level atmospheric forecast variables and geographic variables in a small area centered on a station as predictors. The results show that the SPPM improved the threat score (TS) by 4.29%, 3.66%, 15.63%, 61.08%, and 295.83% for precipitation thresholds of 0.1, 3.0, 10.0, 20.0, and 50.0 mm/3 h, respectively. We then examined the sensitivity of predictors using the interpretable deep-learning technique Layer-wise Relevance Propagation (LRP). The results indicate that the NWP total precipitation (TP) from ECMWF is the most sensitive and important factor, followed by the low-level (850 hPa) field, single-level field, and geographic variables. Notably, TP becomes increasingly important with larger forecast grades, while the importance of variables at other levels remains relatively constant. The majority of stations exhibit consistent importance rankings as mentioned above. Finally, possible causes of variables' insensitivity at medium-level (500 hPa) and high-level (200 hPa) were discussed. • A research idea based on deep learning is offered for short-term station precipitation forecast post-processing. • Training weights based on sample distribution is critical to improving forecasting skills. • Utilizing circulation variables and geographic information fully is vital. • Interpretable methods can demonstrate the reasonable validity of a model. [ABSTRACT FROM AUTHOR]

Abstract Background An attenuated mutant (designated NY303) of Vibrio vulnificus, which causes serious wound infection and septicemia in humans, was isolated fortuitously from a clinical strain YJ016. This mutant was defective in cytotoxicity, migration on soft agar and virulence in the mouse. The purpose of this study was to map the mutation in this attenuated mutant and further explore how the gene thus identified is involved in virulence. Methods The whole genome sequence of mutant NY303 determined by next-generation sequencing was compared with that of strain YJ016 to map the mutations. By isolating and characterizing the specific gene-knockout mutants, the gene associated with the phenotype of mutant NY303 was identified. This gene encodes a global regulator, Lrp. A mutant, YH01, deficient in Lrp was isolated and examined in vitro, in vivo and ex vivo to find the affected virulence mechanisms. The target genes of Lrp were further identified by comparing the transcriptomes, which were determined by RNA-seq, of strain YJ016 and mutant YH01. The promoters bound by Lrp were identified by genome footprinting-sequencing, and those related with virulence were further examined by electrophoretic mobility shift assay. Results A mutation in lrp was shown to be associated with the reduced cytotoxicity, chemotaxis and virulence of mutant NY303. Mutant YH01 exhibited a phenotype resembling that of mutant NY303, and was defective in colonization in the mouse and growth in mouse serum, but not the antiphagocytosis ability. 596 and 95 genes were down- and up-regulated, respectively, in mutant YH01. Many of the genes involved in secretion of the MARTX cytotoxin, chemotaxis and iron-acquisition were down-regulated in mutant YH01. The lrp gene, which was shown to be negatively autoregulated, and 7 down-regulated virulence-associated genes were bound by Lrp in their promoters. A 14-bp consensus sequence, mkCrTTkwAyTsTG, putatively recognized by Lrp was identified in the promoters of these genes. Conclusions Lrp is a global regulator involved in regulation of cytotoxicity, chemotaxis and iron-acquisition in V. vulnificus. Down-regulation of many of the genes associated with these properties may be responsible, at least partly, for loss of virulence in mutant NY303.

Two out of the three major uptake systems for arginine in Escherichia coli are encoded by the artJ-artPIQM gene cluster. ArtJ is the high-affinity periplasmic arginine-specific binding protein (ArgBP-I), whereas artI encodes the arginine and ornithine periplasmic binding protein (AO). Both ArtJ and ArtI are supposed to combine with the inner membrane-associated ArtQMP2 transport complex of the ATP-binding cassette-type (ABC). Transcription of artJ is repressed by arginine repressor (ArgR) and the artPIQM operon is regulated by the transcriptional regulators ArgR and Leucine-responsive regulatory protein (Lrp). Whereas repression by ArgR requires arginine as corepressor, repression of PartP by Lrp is partially counteracted by leucine, its major effector molecule. We demonstrate that binding of dimeric Lrp to the artP control region generates four complexes with a distinct migration velocity, and that leucine has an effect on both global binding affinity and cooperativity in the binding. We identify the binding sites for Lrp in the artP control region, reveal interferences in the binding of ArgR and Lrp in vitro and demonstrate that the two transcription factors act as competitive repressors in vivo, each one being a more potent regulator in the absence of the other. This competitive behavior may be explained by the partial steric overlap of their respective binding sites. Furthermore, we demonstrate ArgR binding to an unusual position in the control region of the lrp gene, downstream of the transcription initiation site. From this unusual position for an ArgR-specific operator, ArgR has little direct effect on lrp expression, but interferes with the negative leucine-sensitive autoregulation exerted by Lrp. Direct arginine and ArgR-dependent repression of lrp could be observed with a 25-bp deletion mutant, in which the ArgR binding site was artificially moved to a position immediately downstream of the lrp transcription initiation site. This finding is reminiscent of a previous observation made for the carAB operon encoding carbamoylphosphate synthase, where ArgR bound in overlap with the downstream promoter P2 does not block transcription initiated 67 bp upstream at the P1 promoter, and further supports the hypothesis that ArgR does not act as an efficient roadblock.

Stimuli of the environment, like objects, systematically activate the actions they are associated to. These activations occur extremely fast. Nevertheless, behavioral data reveal that, in most cases, these activations are then automatically inhibited, around 100 ms after the occurrence of the stimulus. We thus tested whether this early inhibition could be indexed by a central component of the N1 event-related brain potential (ERP). To achieve that goal, we looked at whether this ERP component is larger in tasks that could increase the inhibition and in trials where reaction times (RTs) happen to be long. The illumination of a real space bar of a keyboard out of the dark was used as a stimulus. To maximize the modulation of the inhibition, the task participants had to perform was manipulated across blocks. A look-only task and a count task were used to increase inhibition and an immediate press task was used to decrease it. ERPs of the two block-conditions where presses had to be prevented and where the largest central N1s were predicted were compared to those elicited in the press task, differentiating the ERPs to the third of the trials where presses were the slowest from the ERPs to the third of the trials with the fastest presses. Despite larger negativities due to lateralized readiness potentials (LRPs) and despite greater attention likely in immediate press-trials, central N1s were found to be minimal for the fastest presses, intermediate for the slowest ones and maximal for the two no-press conditions. These results thus provide a strong support for the idea that the central N1 indexes an early and short lasting automatic inhibition of the actions systematically activated by objects. They also confirm that the strength of this automatic inhibition spontaneously fluctuates across trials and tasks. On the other hand, just before N1s, parietal P1s were found larger for fastest presses. They might thus index the initial activation of these actions. Finally, consistent with the idea that N300s index late inhibition processes, that occur preferentially when the task requires them, these ERPs were quasi absent for fast presses trials and much larger in the three other conditions.

Monoclonal antibodies have been explored in a broad range of applications including receptor agonism. Given the importance of receptor conformation in signaling, the agonistic activity of antibodies that engage these receptors are influenced by many parameters. Tetravalent bispecific antibodies that target the frizzled and lipoprotein receptor-related protein receptors and subsequently activate WNT ("Wingless-related integration site" or "Wingless and Int-1" or "Wingless-Int") signaling have been constructed. Because WNT activation stimulates stem cell proliferation and tissue regeneration, immune effector functions should be eliminated from therapeutic antibodies targeting this pathway. Here, we report an unexpected effect of Fc glycosylation on the agonistic activity of WNT mimetic antibodies. Our findings underscore the importance of antibody format, geometry and epitope in agonistic antibody design, and highlight the need to establish appropriate early discovery screening strategies to identify hits for further optimization., Competing Interests: All authors are current or former full-time employees and shareholders of Surrozen, Inc. YL is Executive Vice President of Research at Surrozen, Inc. Patent applications are pending for the work described herein., (© The Author(s) 2024. Published by Oxford University Press on behalf of Antibody Therapeutics.)

Despite the prevalence of visuomotor transformations in our motor skills, their mechanisms remain incompletely understood, especially when imagery actions are considered such as mentally picking up a cup or pressing a button. Here, we used a stimulus-response task to directly compare the visuomotor transformation underlying overt and imagined button presses. Electroencephalographic activity was recorded while participants responded to highlights of the target button while ignoring the second, non-target button. Movement-related potentials (MRPs) and event-related desynchronization occurred for both overt movements and motor imagery (MI), with responses present even for non-target stimuli. Consistent with the activity accumulation model where visual stimuli are evaluated and transformed into the eventual motor response, the timing of MRPs matched the response time on individual trials. Activity-accumulation patterns were observed for MI, as well. Yet, unlike overt movements, MI-related MRPs were not lateralized, which appears to be a neural marker for the distinction between generating a mental image and transforming it into an overt action. Top-down response strategies governing this hemispheric specificity should be accounted for in future research on MI, including basic studies and medical practice., (© The Author(s) 2023. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Systemic toxicity is a major challenge in the development of therapeutics. Consequently, cell-type-specific targeting is needed to improve on-target efficacy while reducing off-target toxicity. Here, we describe a cell-targeting system we have termed BRAID ( BR idged A ctivation by I ntra/intermolecular D ivision) whereby an active molecule is divided into two inactive or less active parts that are subsequently brought together via a so-called 'bridging receptor' on the target cell. This concept was validated using the WNT/β-catenin signaling system, demonstrating that a multivalent WNT agonist molecule divided into two inactive components assembled from different epitopes via the hepatocyte receptor βKlotho induces signaling specifically on hepatocytes. These data provide proof of concept for this cell-specific targeting strategy, and in principle, this may also allow activation of multiple signaling pathways where desirable. This approach has broad application potential for other receptor systems., Competing Interests: HC, SL, NS, TS, CL, YP The authors are current full-time employees and shareholders of Surrozen, Inc, RL, AS, AD, YP, MV The authors were former full-time employees and shareholders of Surrozen, Inc, YL The author is a current full-time employee and shareholder of Surrozen, Inc. YL is Executive Vice President of Research at Surrozen, Inc, (© 2023, Chen, Lee et al.)

Previous studies have analyzed the efficacy of near-infrared spectroscopy-derived lipid core burden index (LCBI) in quantifying and identifying high-risk plaques and patients at increased risk of future major adverse cardiac outcomes/major adverse cardiovascular and cerebrovascular events. A maxLCBI 4mm of 400 or greater seems to be an effective threshold for classifying at-risk plaques. This meta-analysis provides a more precise odds ratio with a narrow standard deviation that can be used to guide future studies., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Munc18-interacting proteins (Mints) are multidomain adaptors that regulate neuronal membrane trafficking, signaling, and neurotransmission. Mint1 and Mint2 are highly expressed in the brain with overlapping roles in the regulation of synaptic vesicle fusion required for neurotransmitter release by interacting with the essential synaptic protein Munc18-1. Here, we have used AlphaFold2 to identify and then validate the mechanisms that underpin both the specific interactions of neuronal Mint proteins with Munc18-1 as well as their wider interactome. We found that a short acidic α-helical motif within Mint1 and Mint2 is necessary and sufficient for specific binding to Munc18-1 and binds a conserved surface on Munc18-1 domain3b. In Munc18-1/2 double knockout neurosecretory cells, mutation of the Mint-binding site reduces the ability of Munc18-1 to rescue exocytosis, and although Munc18-1 can interact with Mint and Sx1a (Syntaxin1a) proteins simultaneously in vitro, we find that they have mutually reduced affinities, suggesting an allosteric coupling between the proteins. Using AlphaFold2 to then examine the entire cellular network of putative Mint interactors provides a structural model for their assembly with a variety of known and novel regulatory and cargo proteins including ADP-ribosylation factor (ARF3/ARF4) small GTPases and the AP3 clathrin adaptor complex. Validation of Mint1 interaction with a new predicted binder TJAP1 (tight junction-associated protein 1) provides experimental support that AlphaFold2 can correctly predict interactions across such large-scale datasets. Overall, our data provide insights into the diversity of interactions mediated by the Mint family and show that Mints may help facilitate a key trigger point in SNARE (soluble N-ethylmaleimide-sensitive factor attachment receptor) complex assembly and vesicle fusion., Competing Interests: Conflict of interest The authors declare that they have no conflicts of interest with the contents of this article., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

Decision making often requires making arbitrary choices ("picking") between alternatives that make no difference to the agent, that are equally desirable, or when the potential reward is unknown. Using event-related potentials we tested the effect of age on this common type of decision making. We compared two age groups: ages 18–25, and ages 41–67 on a masked-priming paradigm while recording EEG and EMG. Participants pressed a right or left button following either an instructive arrow cue or a neutral free-choice picking cue, both preceded by a masked arrow or neutral prime. The prime affected the behavior on the Instructed and the Free-choice picking conditions both in the younger and older groups. Moreover, electrophysiological "Change of Intention" (ChoI) was observed via lateralized readiness potential (LRP) in both age groups – the polarity of the LRP indicated first preparation to move the primed hand and then preparation to move the other hand. However, the older participants were more conservative in responding to the instructive cue, exhibiting a speed-accuracy trade-off, with slower response times, less errors in incongruent trials, and reduced probability of EMG activity in the non-responding hand. Additionally, "Change of Intention" was observed in both age groups in slow RT trials with a neutral prime as a result of an endogenous early intention to respond in a direction opposite the eventual instructing arrow cue. We conclude that the basic behavioral and electrophysiological signatures of implicit ChoI are common to a wide range of ages. However, older subjects, despite showing a similar dynamic decision trajectory as younger adults, are slower, more prudent and finalize the decision making process before letting the information affect the peripheral motor system. In contrast, the flow of information in younger subjects occurs in parallel to the decision process. [ABSTRACT FROM AUTHOR]

Two out of the three major uptake systems for arginine in Escherichia coli are encoded by the artJ - artPIQM gene cluster. ArtJ is the high-affinity periplasmic arginine-specific binding protein (ArgBP-I), whereas artI encodes the arginine and ornithine periplasmic binding protein (AO). Both ArtJ and ArtI are supposed to combine with the inner membrane-associated ArtQMP2 transport complex of the ATP-binding cassette-type (ABC). Transcription of artJ is repressed by arginine repressor (ArgR) and the artPIQM operon is regulated by the transcriptional regulators ArgR and Leucine-responsive regulatory protein (Lrp). Whereas repression by ArgR requires arginine as corepressor, repression of P artP by Lrp is partially counteracted by leucine, its major effector molecule. We demonstrate that binding of dimeric Lrp to the artP control region generates four complexes with a distinct migration velocity, and that leucine has an effect on both global binding affinity and cooperativity in the binding. We identify the binding sites for Lrp in the artP control region, reveal interferences in the binding of ArgR and Lrp in vitro and demonstrate that the two transcription factors act as competitive repressors in vivo , each one being a more potent regulator in the absence of the other. This competitive behavior may be explained by the partial steric overlap of their respective binding sites. Furthermore, we demonstrate ArgR binding to an unusual position in the control region of the lrp gene, downstream of the transcription initiation site. From this unusual position for an ArgR-specific operator, ArgR has little direct effect on lrp expression, but interferes with the negative leucine-sensitive autoregulation exerted by Lrp. Direct arginine and ArgR-dependent repression of lrp could be observed with a 25-bp deletion mutant, in which the ArgR binding site was artificially moved to a position immediately downstream of the lrp transcription initiation site. This finding is reminiscent of a previous observation made for the carAB operon encoding carbamoylphosphate synthase, where ArgR bound in overlap with the downstream promoter P2 does not block transcription initiated 67 bp upstream at the P1 promoter, and further supports the hypothesis that ArgR does not act as an efficient roadblock. [ABSTRACT FROM AUTHOR]

Stimuli of the environment, like objects, systematically activate the actions they are associated to. These activations occur extremely fast. Nevertheless, behavioral data reveal that, in most cases, these activations are then automatically inhibited, around 100 ms after the occurrence of the stimulus. We thus tested whether this early inhibition could be indexed by a central component of the N1 event-related brain potential (ERP). To achieve that goal, we looked at whether this ERP component is larger in tasks that could increase the inhibition and in trials where reaction times (RTs) happen to be long. The illumination of a real space bar of a keyboard out of the dark was used as a stimulus. To maximize the modulation of the inhibition, the task participants had to perform was manipulated across blocks. A look-only task and a count task were used to increase inhibition and an immediate press task was used to decrease it. ERPs of the two block-conditions where presses had to be prevented and where the largest central N1s were predicted were compared to those elicited in the press task, differentiating the ERPs to the third of the trials where presses were the slowest from the ERPs to the third of the trials with the fastest presses. Despite larger negativities due to lateralized readiness potentials (LRPs) and despite greater attention likely in immediate press-trials, central N1s were found to be minimal for the fastest presses, intermediate for the slowest ones and maximal for the two no-press conditions. These results thus provide a strong support for the idea that the central N1 indexes an early and short lasting automatic inhibition of the actions systematically activated by objects. They also confirm that the strength of this automatic inhibition spontaneously fluctuates across trials and tasks. On the other hand, just before N1s, parietal P1s were found larger for fastest presses. They might thus index the initial activation of these actions. Finally, consistent with the idea that N300s index late inhibition processes, that occur preferentially when the task requires them, these ERPs were quasi absent for fast presses trials and much larger in the three other conditions. [ABSTRACT FROM AUTHOR]

No consensus has been attained regarding the utility of open retropubic radical prostatectomy (RRP), laparoscopic radical prostatectomy (LRP) and robot-assisted laparoscopic radical prostatectomy (RALRP) for localized prostate cancer (PCa). We carried out a network meta-analysis and cumulative meta-analysis comparing RRP, LRP and RALRP on peri-operative and functional outcome measures. Electronic databases were searched for either randomized clinical trials or cohort studies comparing RALRP either with LRP or RRP in patients with localized PCa. Outcome measures were as follows: overall, pT2 and pT3-positive surgical margins (PSMs); biochemical recurrence (BCR); complication rates; estimated blood loss; blood transfusion rate; continence and potency rates; duration of catheterization and hospital stay. Publication bias, risk of bias and inconsistency were assessed. Inverse heterogeneity model was used for analysis. A total of 45 studies were included for the final analysis. We observed that RALRP and LRP did not differ significantly from RRP with regard to the following outcomes: overall PSM; pT2 and pT3 PSMs; OT; complication rate; continence and potency rates; total blood loss and hospital stay. Duration of catheterization was significantly shorter in RALRP than LRP and RRP while significant reductions in the need for blood transfusion and BCR were observed for both RALRP and LRP in comparison with RRP. To conclude, similar functional, operative and oncologic outcomes were observed for both RALRP and LRP compared to RRP. [ABSTRACT FROM AUTHOR]

Highlights • Facial gender and attractiveness: Key features to study face processes. • Processing onsets estimated by LRP- and N200-effects. • Attractiveness follows gender in terms of underlying neural processes. • Early face attractiveness assessment seems to rely on gender-stereotypes. • Used paradigm is promising to further uncover the time course of face perception. Abstract We followed an ERP-based approach to gain knowledge on the dependence and temporal order of two essential processes of face perception: attractiveness and gender. By combining a dual-choice task with a go/nogo-paradigm focusing on the LRP and N200-effect, we could estimate the processing times and onsets of both types of face processing. The analyses of the LRP revealed that gender aspects were processed much earlier than attractiveness. Whereas gender was already analysed 243.9 ms post-stimulus onset, attractiveness came into play 58.6 ms later, i.e. after a post-stimulus onset delay of 302.5 ms. This resulting pattern was mirrored by the analyses of the N200-effect, an effect available mainly frontally which is supposed to correlate with the inhibition of inappropriate responses. Taking the onset of the N200 effect as an estimator for the moment at which information has been processed sufficiently for task decision, we could trace the N200 effect at 152.0 ms for go/nogo-decision on gender, while not as early as 206.7 ms on attractiveness. In sum, processing of facial attractiveness seems to be based on gender-specific aesthetic pre-processing, for instance via activating gender-specific attractiveness prototypes which show focused processing of certain facial aspects. [ABSTRACT FROM AUTHOR]

The human pathogen, Vibrio cholerae, belongs to the 10% of bacteria in which the genome is divided. Each of its two chromosomes, like bacterial chromosomes in general, replicates from a unique origin at fixed times in the cell cycle. Chr1 initiates first, and upon duplication of a site in Chr1, crtS, Chr2 replication initiates. Recent in vivo experiments demonstrate that crtS binds the Chr2-specific initiator RctB and promotes its initiator activity by remodeling it. Compared to the well-defined RctB binding sites in the Chr2 origin, crtS is an order of magnitude longer, suggesting that other factors can bind to it. We developed an in vivo screen to identify additional crtS-binding proteins and identified the global transcription factor, Lrp, as one such protein. Studies in vivo and in vitro indicate that Lrp binds to crtS and facilitates RctB binding to crtS. Chr2 replication is severely defective in the absence of Lrp, indicative of a critical role of the transcription factor in licensing Chr2 replication. Since Lrp responds to stresses such as nutrient limitation, its interaction with RctB presumably sensitizes Chr2 replication to the physiological state of the cell.

The most important causes of treatment failure in canine lymphoma include intrinsic or acquired drug resistance. Thus, elucidation of molecular mechanisms of drug resistance is essential for the establishment of better treatment alternatives for lymphoma patients. The overexpression of drug transporters is one of the most intensively studied mechanisms of drug resistance in many tumors. In canine lymphoma, it has also been shown that the overexpression of drug efflux pumps such as P-glycoprotein is associated with drug-resistant phenotypes. Canine lymphoma has many pathological similarities to human non-Hodgkin’s lymphoma, and they also share similar molecular mechanisms of drug resistance. We have previously demonstrated the association of the overexpression of drug transporters with drug resistance and indicated some molecular mechanisms of the regulation of these transporters’ expressions in canine and human lymphoid tumor cells. However, it has also been indicated that other known or novel drug resistance factors should be explored to overcome drug resistance in lymphoma. In this review, we summarize the recent findings on the molecular mechanisms of drug resistance and possible strategies to develop better treatment modalities for canine lymphoma from the comparative aspects with human lymphoid tumors.

Abstract Why do individuals fail to exercise regularly despite knowledge of the risks associated with physical inactivity? Automatic processes regulating exercise behaviors may partly explain this paradox. Yet, these processes have only been investigated with behavioral outcomes (i.e., based on reaction times). Here, using electroencephalography, we investigated the cortical activity underlying automatic approach and avoidance tendencies toward stimuli depicting physical activity and sedentary behaviors in 29 young adults who were physically active or physically inactive but with the intention of becoming physically active. Behavioral results showed faster reactions when approaching physical activity compared to sedentary behaviors and when avoiding sedentary behaviors compared to physical activity. These faster reactions were more pronounced in physically active individuals and were associated with changes during sensory integration (earlier onset latency and larger positive deflection of the stimulus-locked lateralized readiness potentials) but not during motor preparation (no effect on the response-locked lateralized readiness potentials). Faster reactions when avoiding sedentary behaviors compared to physical activity were also associated with higher conflict monitoring (larger early and late N1 event-related potentials) and higher inhibition (larger N2 event-related potentials), irrespective of the usual level of physical activity. These results suggest that additional cortical resources were required to counteract an attraction to sedentary behaviors. Data and Materials [ https://doi.org/10.5281/zenodo.1169140 ]. Preprint [ https://doi.org/10.1101/277988 ]. Highlights • Individuals are slower at approaching sedentary than physical activity stimuli. • Individuals are quicker at avoiding sedentary than physical activity stimuli. • These effects are particularly pronounced in physically active individuals. • Avoiding sedentary behaviors is associated with high levels of conflict monitoring and inhibition. • Additional brain resources are required to escape a general attraction toward sedentary behaviors. [ABSTRACT FROM AUTHOR]

The human pathogen, Vibrio cholerae , belongs to the 10% of bacteria in which the genome is divided. Each of its two chromosomes, like bacterial chromosomes in general, replicates from a unique origin at fixed times in the cell cycle. Chr1 initiates first, and upon duplication of a site in Chr1, crtS , Chr2 replication initiates. Recent in vivo experiments demonstrate that crtS binds the Chr2-specific initiator RctB and promotes its initiator activity by remodeling it. Compared to the well-defined RctB binding sites in the Chr2 origin, crtS is an order of magnitude longer, suggesting that other factors can bind to it. We developed an in vivo screen to identify additional crtS -binding proteins and identified the global transcription factor, Lrp, as one such protein. Studies in vivo and in vitro indicate that Lrp binds to crtS and facilitates RctB binding to crtS. Chr2 replication is severely defective in the absence of Lrp, indicative of a critical role of the transcription factor in licensing Chr2 replication. Since Lrp responds to stresses such as nutrient limitation, its interaction with RctB presumably sensitizes Chr2 replication to the physiological state of the cell. [ABSTRACT FROM AUTHOR]

Poor Mg status is a risk factor for Alzheimer’s disease (AD), and the underlying mechanisms remain elusive. Here, we provided the first evidence that elevated Mg levels significantly reduced the blood-brain barrier (BBB) permeability and regulated its function in vitro. Transient receptor potential melastatin 7 (TRPM7) and magnesium transporter subtype 1 (MagT1) were two major cellular receptors mediating entry of extracellular Mg2+ into the cells. Elevated Mg levels also induced an accelerated clearance of amyloid-β peptide (Aβ) from the brain to the blood side via BBB transcytosis through low-density lipoprotein receptor-related protein (LRP) and phosphatidylinositol binding clathrin assembly protein (PICALM), while reduced the influx of Aβ from the blood to the brain side involving receptor for advanced glycation end products (RAGE) and caveolae. Mg enhanced BBB barrier properties and overall expression of LRP1 and PICALM whereas reduced that of RAGE and caveolin-1. Apical-to-basolateral and vice versa steady-state Aβ flux achieved an equilibrium of 18 and 0.27 fmol/min/cm2, respectively, about 30 min after the initial addition of physiological levels of free Aβ. Knockdown of caveolin-1 or disruption of caveolae membrane microdomains reduced RAGE-mediated influx significantly, but not LRP1-mediated efflux of Aβ. Stimulating endothelial cells with vascular endothelial growth factor (VEGF) enhanced caveolin-1 phosphorylation and RAGE expression. Co-immunoprecipitation demonstrated that RAGE, but not LRP1, was physically associated with caveolin-1. Thus, Mg can reduce BBB permeability and promote BBB clearance of Aβ from the brain by increasing the expression of LRP1 and PICALM while reducing the level of RAGE and caveolin-1. [ABSTRACT FROM AUTHOR]

In collaborative logistics network, all task demands are centralized by logistics platform for resource allocation, and then resource owners execute discretely under final plan. The key point of this study is to effectively balance the different appeals of stakeholders. In this paper, a bi-level programming model is presented to solve multi-deport LRP considered the constraint of hard time window, vehicle capacity and vehicle backhaul cost, which aims at achieving the effective interest coordination between upper layer platform and lower layer vehicle owners. To solve this bi-level model, genetic algorithm is redesigned according to this specific situation. Optimal results of simulation sample show that the satisfied solution for both platform and vehicle owners could be obtained using this method. Feasibility and validity of this model and adaptive algorithm have been verified in the paper. The bi-level model provides a practical and effective method to solve the profit distribution problem between platform and vehicle owners. [ABSTRACT FROM AUTHOR]

The cellular mechanisms that induce calcific aortic stenosis are yet to be unraveled. Wnt signaling is increasingly being considered as a major player in the disease process. However, the presence of Wnt Frizzled (Fzd) receptors and co-receptors LRP5 and 6 in normal and diseased human aortic valves remains to be elucidated. Immunohistochemistry and quantitative polymerase chain reaction were used to determine Fzd receptor expression in normal and calcified human aortic valve tissue, as well as human aortic valve interstitial cells (HAVICs) isolated from calcified and normal human aortic valves. There was significantly higher mRNA expression of 4 out of the 10 Fzd receptors in calcified aortic valve tissues and 8 out of the 10 in HAVICs, and both LRP5/6 co-receptors in calcified aortic valves ( P < 0.05). These results were confirmed by immunohistochemistry, which revealed abundant increase in immunoreactivity for Fzd3, 7, and 8, mainly in areas of lipid core and calcified nodules of diseased aortic valves. The findings of abundant expression of Fzd and LRP5/6 receptors in diseased aortic valves suggests a potential role for both canonical and noncanonical Wnt signaling in the pathogenesis of human aortic valve calcification. Future investigations aimed at targeting these molecules may provide potential therapies for aortic valve stenosis. [ABSTRACT FROM AUTHOR]