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With the application of hyperspectral imaging systems on high-throughput plant phenotyping, multivariable spectral data modelling methods have been proposed to predict plant physiological features such as biomass. However, most presented modelling methods are linear models such as the regression model between projected leaf area and plant biomass, which cannot accurately reflect some non-linear relationships between hyperspectral imaging information and the phenomes to be predicted. Therefore, it is important to develop a prediction model for plant physiological features that fully utilises the information in plant hyperspectral images. In this paper, a non-linear modelling method known as kernel partial least squares (KPLS) was investigated to improve the prediction performance of maize (US corn) biomass. The main idea of KPLS is mapping the original input features into higher dimensional feature spaces before obtaining the principle components for a standard PLS model. In the new mapping spaces, the original, nonlinear inputs may exhibit linear patterns with different kernel functions. This method was tested in a greenhouse assay containing 102 maize plants, which were subjected to a combination of three water treatments and two nitrogen treatments at three different growth stages. KPLS methods were successfully applied on the collected hyperspectral images. The results showed that compared with conventional linear models, statistics including R2 of KPLS with radial basis function (RBF) kernel had improved accuracy (R2 = 0.924). We expect that besides plant biomass, this new non-linear KPLS model could improve the quality of estimation of other plant features as well.

A novel lipopeptide antibiotic, stalobacin I (1), was discovered from a culture broth of an unidentified Gram-negative bacterium. Stalobacin I (1) had a unique chemical architecture composed of an upper and a lower half peptide sequence, which were linked via a hemiaminal methylene moiety. The sequence of 1 contained an unusual amino acid, carnosadine, 3,4-dihydroxyariginine, 3-hydroxyisoleucine, and 3-hydroxyaspartic acid, and a novel cyclopropyl fatty acid. The antibacterial activity of 1 against a broad range of drug-resistant Gram-positive bacteria was much stronger than those of "last resort" antibiotics such as vancomycin, linezolid, and telavancin (MIC 0.004-0.016 μg/mL). Furthermore, compound 1 induced a characteristic morphological change in Gram-positive and Gram-negative strains by inflating the bacterial cell body. The absolute configuration of a cyclopropyl amino acid, carnosadine, was determined by the synthetic study of its stereoisomers, which was an essential component for the strong activity of 1.

Grammar acquisition by non-native learners (L2) is typically less successful and may produce fundamentally different grammatical systems than that by native speakers (L1). The neural representation of grammatical processing between L1 and L2 speakers remains controversial. We hypothesized that working memory is the primary source of L1/L2 differences, and operationalized working memory is an active inference within the predictive coding account, which models grammatical processes as higher-level neuronal representations of cortical hierarchies, generating predictions (forward model) of lower-level representations. A functional MRI study was conducted with L1 Japanese speakers and highly proficient Japanese learners requiring oral production of grammatically correct Japanese particles. Selecting proper particles requires forward model-dependent active inference as their functions are highly context-dependent. As a control, participants read out a visually designated mora indicated by underlining. Particle selection by L1/L2 groups commonly activated the bilateral inferior frontal gyrus/insula, pre-supplementary motor area, left caudate, middle temporal gyrus, and right cerebellum, which constituted the core linguistic production system. In contrast, the left inferior frontal sulcus, known as the neural substrate of verbal working memory, showed more prominent activation in L2 than in L1. Thus, the active inference process causes L1/L2 differences even in highly proficient L2 learners.

Invasive fungal infections (IFIs) are fatal infections, but treatment options are limited. The clinical efficacies of existing drugs are unsatisfactory because of side effects, drug-drug interaction, unfavorable pharmacokinetic profiles, and emerging drug-resistant fungi. Therefore, the development of antifungal drugs with a new mechanism is an urgent issue. Herein, we report novel aryl guanidine antifungal agents, which inhibit a novel target enzyme in the ergosterol biosynthesis pathway. Structure-activity relationship development and property optimization by reducing lipophilicity led to the discovery of 6h, which showed potent antifungal activity against Aspergillus fumigatus in the presence of serum, improved metabolic stability, and PK properties. In the murine systemic A. fumigatus infection model, 6h exhibited antifungal efficacy equivalent to voriconazole (1e). Furthermore, owing to the inhibition of a novel target in the ergosterol biosynthesis pathway, 6h showed antifungal activity against azole-resistant A. fumigatus.

Coadministration of β-lactam and β-lactamase inhibitor (BLI) is one of the well-established therapeutic measures for bacterial infections caused by β-lactam-resistant Gram-negative bacteria, whereas we have only two options for orally active BLI, clavulanic acid and sulbactam. Furthermore, these BLIs are losing their clinical usefulness because of the spread of new β-lactamases, including extended-spectrum β-lactamases (ESBLs) belonging to class A β-lactamases, class C and D β-lactamases, and carbapenemases, which are hardly or not inhibited by these classical BLIs. From the viewpoints of medical cost and burden of healthcare personnel, oral therapy offers many advantages. In our search for novel diazabicyclooctane (DBO) BLIs possessing a thio-functional group at the C2 position, we discovered a 2-sulfinyl-DBO derivative (2), which restores the antibacterial activities of an orally available third-generation cephalosporin, ceftibuten (CTB), against various serine β-lactamase-producing strains including carbapenem-resistant Enterobacteriaceae (CRE). It can be orally absorbed via the ester prodrug modification and exhibits in vivo efficacy in a combination with CTB.

Objective: The epidemiology and role of the anti-PcrV titer in non-cystic fibrosis patients with Pseudomonas aeruginosa airway tract infections is not fully understood. This study was performed to compare the anti-PcrV titers of patients with and without P. aeruginosa respiratory tract infections. Methods: This prospective cohort study was conducted at Hokkaido University Hospital in Japan. Participants had blood and sputum specimens collected on admission. They were divided into two groups based on their sputum culture results. Those with a P. aeruginosa infection were assigned to the P. aeruginosa (PA) group and those without a P. aeruginosa infection were assigned to the non-PA group. Serum anti-PcrV titers were measured using a validated ELISA. Results: Of the 44 participants, 15 were assigned to the PA group and 29 were assigned to the non-PA group. In the PA group, 10/15 participants (66.7%) had an anti-PcrV titer >1000 ng/ml compared to 3/29 participants (10.3%) in the non-PA group (p < 0.001). In the PA group, two of the five participants with an anti-PcrV titer

Greenhouse plant science assays have been impacted by microclimates which causes significant level of noise to plant growth measurement data. Researchers and scientist have been randomizing pots locations, which helps to re-distribute the noise, but does not remove the noise. The impacts from microclimates can be eliminated by shuffling plants, but there has been no study on the optimization of shuffling pattern, such as the frequency and moving distance for the pots. It is important to quantitatively study the microclimates in the greenhouse, so we can optimize the shuffling pattern accordingly. The aim of this study was to propose a computer modeling approach for simulating microclimate in the greenhouse, and then use the simulation result to optimize pot movement distance and frequency. A computational greenhouse model was developed using inputs from real design, materials and location of a Purdue Lily greenhouse in West Lafayette, Indiana. Microclimate variables, including ambient temperature and lighting radiation over 24 h and 7 days were predicted with the simulation model. Thermometers and lighting sensors were also distributed in the greenhouse for the ground-truth measurements over a seven-day period. Comparison of measured microclimate variables with predicted variables obtained from the computational model demonstrated that the simulation model could precisely predict temperatures and light radiation at any time, and at different positions in the greenhouse. Optimized pot movement frequency and distance were then determined with the simulation result. The new shuffling pattern can remove over 90% of the microclimate variance but could save more than 95% shuffling efforts compared with non-stop movement.

During recent years, hyperspectral imaging systems have been widely applied in the greenhouses for plant phenotyping purposes. Current systems are typically designed as either top view or side view imaging mode. Top view is an ideal imaging angle for top leaves with flat leaf surfaces. However, most bottom leaves are either blocked or shaded. From side view, the entire plant structure is viewable. However, most leaf surfaces are not facing the camera, which impacts measurement quality. Besides, there could be advantages with certain tilted angle(s) between top view and side view. It’s interesting to explore the impact of different imaging angles to the phenotyping quality. For this purpose, a swing hyperspectral imaging system capable of capturing images at any angle from side view (0°) to top view (90°) by rotating the camera and the lighting source was designed. Corn plants were grown and allocated into 3 different treatments: high nitrogen (N) and well-watered (control), high N and drought-stressed, and low N and well-watered. Each plant was imaged at 7 different angles from 0° to 90° with an interval of 15°. The soil plant analysis development (SPAD) values and relative water content (RWC) ground truth measurements were used to establish treatment effects. The results showed that averaged plant-level Normalized Difference Vegetation Index (NDVI) values of plants in different treatments changed at different imaging angles. The results also indicated that for pixel-level NDVI distributions, the titled imaging angle of 75° was optimal to distinguish different water treatments, whereas, the tilted imaging angle of 15° was optimal to distinguish different N treatments. For pixel-level RWC distributions, the distribution difference between different water treatments was larger at higher imaging angles.

Background: The number of bacterial infections that—for various reasons—are challenging to cure continues to increase. One such reason is persister cell infection. To investigate persister formation and persister infections, viable bacteria must be evaluated in the same animal over time. In this study, the feasibility of monitoring viable bacteria by SPECT using two labeled peptides was evaluated. Results: Two types of ubiquicidin (UBI) 29-41 labeled with technetium-99m, 99mTc-HYNIC(GH)2-UBI 29-41 and 99mTc-HYNIC(Tricine)2-UBI 29-41, were synthesized. The in vitro binding of these labeled peptides to Staphylococcus aureus was measured. For the in vivo study, each labeled peptide was injected into S. aureus infected mouse thigh after treatment with various doses of ciprofloxacin (CPFX). Two hours after injection, the accumulation of each labeled peptide at the infection site was assessed by SPECT, and then the number of viable bacteria was determined from the accumulation detected. The peptide labeling was successful, and the radiochemical purity was 91±9% (GH, n=8) and 100% (Tricine, n=8). The in vitro binding of the labeled peptides to S. aureus (5×108 cfu) without serum was 78.9% (GH) and 85.5% (Tricine) of the total 99mTc activity. With serum, the binding rate was 67.5% (GH) and 13.3% (Tricine). The accumulation of labeled peptide was calculated from the SPECT images, and that in the bacterial infection site (left thigh) was higher than that in the non-infection site (right thigh) for both peptides. Good correlation was found between the target-to-non-target (T/NT) ratios of each labeled peptide and the viable bacterial count at the infection site, and 99mTc-HYNIC(Tricine)2-UBI 29-41 had a wider range than 99mTc-HYNIC(GH)2-UBI 29-41.Conclusion: Using the SPECT/labeled peptide method, it was possible to monitor viable bacterial count in the range 103–108 cfu, which is appropriate for tracking viable bacterial counts in the same animal over time.

Potent fungicidal activity is one of the key factors of antifungals to overcome invasive pulmonary aspergillosis (IPA). To date, quantification of Aspergillus DNA in the lungs and galactomannan (GM) in serum or bronchoalveolar lavage fluid have been developed as general methods for measuring fungal burden in IPA animal models. However, GM quantification is not supposed to be a suitable method for precise evaluation of the fungicidal effects of antifungals, because killed Aspergillus hyphae can release GM for a certain period until they are eliminated by the host. Therefore, in terms of detecting viable fungal burden of Aspergillus, quantification of Aspergillus DNA has been thought to be a suitable method. Here, to obtain a method with much higher sensitivity, we applied reverse transcription quantitative PCR (RT-qPCR) for A. fumigatus 18S ribosomal RNA to measure the viable fungal burden in murine IPA models. Prior to in vivo tests, we confirmed that the sensitivity of 18S rRNA was nearly 50-fold higher than that of 18S ribosomal DNA in vitro. This highly sensitive method made it possible to evaluate the fungicidal effects of antifungals in a low-inoculation murine IPA model. In this model, single administrations of higher doses of voriconazole and posaconazole, which have fungicidal activity, were able to display fungicidal effects with ≥1 log10 reductions by 18S rRNA quantification, whereas significant reductions in serum GM were not observed. These results suggest that 18S rRNA quantification is a powerful tool for screening novel antifungals with potent fungicidal activity only after a single administration.

Objective To investigate possible mechanistic factors to explain cefiderocol (CFDC) non-susceptibility, we characterized 38 clinical isolates with a CFDC minimum inhibitory concentration (MIC) of >4 μg/mL from a multi-national surveillance study. Methods The MIC measurement in the presence of β-lactamase inhibitors and whole genome sequencing were performed. Results The MIC decrease of CFDC by β-lactamase inhibitors was observed against all of the test isolates. Among the 38 isolates, NDM and PER genes were observed in 5 and 25 isolates, respectively. No other β-lactamases responsible for high MIC were identified in the other eight isolates. The MIC of CDFC against Escherichia coli isogenic strains introduced with NDM and PER β-lactamase increased by ≥16-fold, suggesting the contribution of NDM and PER to the non-susceptibility to CFDC. Against NDM producers, a ≥8-fold MIC increase was observed only when both serine- and metallo-type β-lactamase inhibitors were added. In addition, many of the PER or NDM producers remained susceptible to CFDC. These results suggested that the presence of only NDM or PER would not lead to non-susceptibility to CFDC and that multiple factors would be related to CFDC resistance. Conclusion Multiple factors including NDM and PER could be related to reduced susceptibility to CFDC.

Aerial imaging technologies have been widely applied in agricultural plant remote sensing. However, an as yet unexplored challenge with field imaging is that the environmental conditions, such as sun angle, cloud coverage, temperature, and so on, can significantly alter plant appearance and thus affect the imaging sensor’s accuracy toward extracting plant feature measurements. These image alterations result from the complicated interaction between the real-time environments and plants. Analysis of these impacts requires continuous monitoring of the changes through various environmental conditions, which has been difficult with current aerial remote sensing systems. This paper aimed to propose a modeling method to comprehensively understand and model the environmental influences on hyperspectral imaging data. In 2019, a fixed hyperspectral imaging gantry was constructed in Purdue University’s research farm, and over 8000 repetitive images of the same corn field were taken with a 2.5 min interval for 31 days. Time-tagged local environment data, including solar zenith angle, solar irradiation, temperature, wind speed, and so on, were also recorded during the imaging time. The images were processed for phenotyping data, and the time series decomposition method was applied to extract the phenotyping data variation caused by the changing environments. An artificial neural network (ANN) was then built to model the relationship between the phenotyping data variation and environmental changes. The ANN model was able to accurately predict the environmental effects in remote sensing results, and thus could be used to effectively eliminate the environment-induced variation in the phenotyping features. The test of the normalized difference vegetation index (NDVI) calculated from the hyperspectral images showed that variance in NDVI was reduced by 79%. A similar performance was confirmed with the relative water content (RWC) predictions. Therefore, this modeling method shows great potential for application in aerial remote sensing applications in agriculture, to significantly improve the imaging quality by effectively eliminating the effects from the changing environmental conditions.

Pseudomonas aeruginosa is one of the most common and clinically important pathogens because of its resistance to a wide variety of antibiotics. A number of treatments of P. aeruginosa have been developed, but there is still no definitive one. Antisense drugs have a great potential to treat multidrug-resistant P. aeruginosa because this technology, in principle, can inhibit the expression of any essential genes. Nucleic Acid Ther.2012, 22, 323 reported that peptide nucleic acid (PNA) antisenses conjugated to the carrier peptide (RXR)4 and targeted to ftsZ and acpP (essential genes) had antibacterial activity in P. aeruginosa. However, growth inhibition was also found with peptide-PNA antisense conjugates of mismatched sequences (negative controls), and hence there remains a possibility for considerable enhancement of basal level activity due to the general toxicity. To assess the true potential of peptide-PNA conjugates, we measured sequence-dependent knockdown of the (RXR)4-PNA conjugates by using a scrambled sequence as a negative control. In addition, we evaluated (RXR)4-PNA antisenses against three other essential genes (lepB, lptD and mraY) and a non-essential gene (PA1303), and confirmed that multiple sequences targeting only the essential genes showed antimicrobial activity in P. aeruginosa PAO1 cells. We also conducted a rescue experiment and confirmed that the antimicrobial activity of anti-mraY antisenses was an on-target effect, not due to general toxicity. These findings indicate that the (RXR)4–PNA antisense should be a useful tool for target validation of a specific gene and could be a therapeutic platform capable of targeting a variety of genes in P. aeruginosa.

The analysis of the rhythm of leaf movement is a simple yet effective method to quantify the impacts of external (e.g. abiotic stress) and/or internal (e.g. gene mutations) perturbations on plant growth. We developed an automated monitoring system to quantify leaf movement using time-lapse imaging and a subsequent leaf-tracking algorithm. The leaf-tracking algorithm was based on dense optical flow algorithm to directly record temporal motion events. The algorithm measures motion directly, rather than detecting leaf or cotyledon tip in every image, so multiple leaves, including occluded leaves, can be measured simultaneously. To test the monitoring system, wild-type and drought-tolerant mutant genotypes of Arabidopsis (Arabidopsis thaliana) were subjected to a combinatorial two water and two nitrogen levels. High-frequency time-lapse images were acquired from top view for little over 6 consecutive days at a frequency of 4 min. Results showed that nitrogen and water treatments elicited differences in mean plant displacement in both genotypes. It also showed significant differences among the two different genotypes in the mean displacement when plants were under water or nitrogen stress. These results confirmed the new monitoring system’s ability to discern environmental and genotypic differences in plant response.

Aspergillus flavus is the second most significant pathogenic cause of invasive aspergillosis; however, its emergence risks and mechanisms of voriconazole (VRC) resistance have not yet been elucidated in detail. Here, we demonstrate that repeated exposure of A. flavus to subinhibitory concentrations of VRC in vitro causes the emergence of a VRC-resistant mutant with a novel resistance mechanism. The VRC-resistant mutant shows a MIC of 16 μg/ml for VRC and of 0.5 μg/ml for itraconazole (ITC). Whole-genome sequencing analysis showed that the mutant possesses a point mutation in yap1, which encodes a bZIP transcription factor working as the master regulator of the oxidative stress response, but no mutations in the cyp51 genes. This point mutation in yap1 caused alteration of Leu558 to Trp (Yap1(Leu558Trp)) in the putative nuclear export sequence in the carboxy-terminal cysteine-rich domain of Yap1. This Yap1(Leu558Trp) substitution was confirmed as being responsible for the VRC-resistant phenotype, but not for that of ITC, by the revertant to Yap1(wild type) with homologous gene replacement. Furthermore, Yap1(Leu558Trp) caused marked upregulation of the atrF ATP-binding cassette transporter, and the deletion of atrF restored susceptibility to VRC in A. flavus. These findings provide new insights into VRC resistance mechanisms via a transcriptional factor mutation that is independent of the cyp51 gene mutation in A. flavus.

To compare the risk of acquiring in vitro resistance between doripenem and tazobactam/piperacillin by CTX-M-15-producing Escherichia coli, the in vitro frequency of resistance was determined. Four strains carrying multiple β-lactamases such as blaOXA-1 or blaCTX-M-27 as well as blaCTX-M-15 and blaTEM-1 were used. No resistant colonies appeared on doripenem-containing plates, whereas resistant colonies were obtained from three of four test strains against tazobactam/piperacillin using agar plate containing 8- to 16-fold MIC of each drug. These three acquired tazobactam/piperacillin-resistant strains were not cross-resistant to doripenem, and they showed 1.9- to 3.1-fold higher piperacillin-hydrolysis activity compared to those of each parent strain. The change of each β-lactamase mRNA expression measured by real-time PCR varied among three resistant strains. One of three tazobactam/piperacillin-resistant strains with less susceptibility to ceftazidime overexpressed both blaCTX-M-15 and blaTEM-1, and the other two strains showed higher mRNA expression of either blaTEM-1 or blaOXA-1. These results demonstrate that multiple β-lactamases carried by CTX-M-15-producing E. coli contributed to the resistance to tazobactam/piperacillin. On the other hand, these resistant strains maintained susceptibility to doripenem. The risk of acquiring in vitro resistance to doripenem by CTX-M-15-producing E. coli seems to be lower than that to tazobactam/piperacillin.

A 56-year-old man experienced a sudden onset of left hemiparesis. The computed tomography(CT)scan revealed a lobar hemorrhage in the right fronto-parietal lobe. After his admission, deep vein thrombosis was detected in his left lower limb, and angiograms taken on the 36th day of hospitalization revealed cerebral venous sinus thrombosis. Anticoagulant treatment was induced. After 21 months, he experienced a sudden onset of left hemiparesis again. The CT scan revealed a new lobar hemorrhage in the right frontal lobe, and angiograms revealed that two dural arteriovenous fistulas(dAVF)developed in the superior sagittal sinus(SSS)and the left transverse-sigmoid sinus. The one in the SSS had retrograde drainage from the bilateral middle meningeal artery, and we performed transarterial embolization with 50% n-butyl-cyanoacrylate. Postoperative course was uneventful and no further stroke occurred. Intracranial dAVF is known to be an acquired disease caused by venous hypertension, but we rarely find new development of dAVFs after venous diseases. This patient's case will help to elucidate the pathophysiology of dAVF.

We developed a surface plasmon resonance (SPR) assay to estimate the interactions of antimicrobial agents with the dipeptide terminal of lipid II ( d -alanyl- d -alanine) and its analogous dipeptides ( l -alanyl- l -alanine and d -alanyl- d -lactate) as ligands. The established SPR method showed the reproducible immobilization of ligands on sensor chip and analysis of binding kinetics of antimicrobial agents to ligands. The ligand-immobilized chip could be used repeatedly for at least 200 times for the binding assay of antimicrobial agents, indicating that the ligand-immobilized chip is sufficiently robust for the analysis of binding kinetics. In this SPR system, the selective and specific binding characteristics of vancomycin and its analogs to the ligands were estimated and the kinetic parameters were calculated. The kinetic parameters revealed that one of the remarkable binding characteristics was the specific interaction of vancomycin to only the d -alanyl- d -alanine ligand. In addition, the kinetic binding data of SPR showed close correlation with the antimicrobial activity. The SPR data of other antimicrobial agents (e.g., teicoplanin) to the ligands showed correlation with the antimicrobial activity on the basis of the therapeutic mechanism. Our SPR method could be a valuable tool for predicting the binding characteristics of antimicrobial agents to the dipeptide terminal of lipid II.