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This study aims to analyze time-series measurements encompassing rainstorm events with over a century of datasets to identify rainstorm evolution and dimensional transitions in non-stationarity. Rainstorm events are identified using partial duration series (PDS) to extract changes in rainstorm characteristics, namely maximum intensity (MAXI), duration (D), total rainfall (TR), and average rainfall intensity (ARI), in response to climate change. Ensemble empirical mode decomposition is used for trend filtering and non-stationary identification to explore spatiotemporal insight patterns. Trend models for the first-second-order moments of rainstorm characteristics are used to formulate the identified mean-variance trends using combined multi-dimensional linear-parabolic regression. Best-fitting combinations of various distributions (probability density functions) and trend models for multiple characteristic series are identified based on the Akaike information criterion. We analyze the dimensional transition in rainfall non-stationarity based on sensitivity analysis using PDS to determine its natural geophysical causes. The integrated methodology was applied to the data retrieved from nine weather stations in Taiwan. Our findings reveal rainstorm characteristics of "short D but high rainfall intensity" or "lower MAXI but high TR" across multiple stations. The parabolic trend of the first-order moment (i.e., mean) of ARI, D, and TR appears at the endpoint of the mountain ranges. Areas receiving monsoons and those on the windward plain show a rising parabolic trend in the first- and second-order moments (i.e., mean-variance) characterizing MAXI, implying that the occurrence frequency and magnitude of extreme MAXI increases. Non-stationary transitions in MAXI appear for mountain ranges exposed to the monsoon co-movement effect on both windward and leeward sides. Stations in the plains and rift valleys show upgraded and downgraded transitions in the non-stationary dimensions for D, respectively., (© 2024. The Author(s).)

Misfolded protein oligomers are of central importance in both the diagnosis and treatment of Alzheimer's and Parkinson's diseases. However, accurate high-throughput methods to detect and quantify oligomer populations are still needed. We present here a single-molecule approach for the detection and quantification of oligomeric species. The approach is based on the use of solid-state nanopores and multiplexed DNA barcoding to identify and characterize oligomers from multiple samples. We study α-synuclein oligomers in the presence of several small-molecule inhibitors of α-synuclein aggregation as an illustration of the potential applicability of this method to the development of diagnostic and therapeutic methods for Parkinson's disease.

To precisely identify multi-dimensional spatiotemporal rain-making parameters, generate an approximate Hessian matrix, and solve the nonlinear ill-posed problem, this study uses composite logical tangent hyperbolic functions to construct the rain-generating simulation model as nonlinear algebraic equations with designed key-lock quasi-Newton optimization for deriving multi-order objective functional derivatives for rainstorm causal decomposition into advanced functional, analytical solution (lock) and Newton's conditional constraints. Specifically, the rank-two approximate structure of the Levenberg-Marquardt and Broyden-Fletcher-Goldfarb-Shanno quasi-Newton algorithms are modified as the symmetric rank-four structure to efficiently calculate a positive definite stable Hessian and solve the constrained nonlinear rain-making threshold. The model projects various rain-making factors into multi-rank loading scores, characterizing rain-generating mechanisms and causal components as associated DNAs. To accelerate/modify directional convergence, avoid local minimum, and detect global optimum, the devised vectorized limited switchable step sizes are optimized using advanced double-bracketing approaches combined with candidate parameters' correction vectors (key) and referenced step-size distributions solved by Newton's constrained analytical solution to reduce heterogeneous differences and eliminate the conventional overestimated Hessian. The identified rain-making DNAs reveal that typhoons with similar DNAs move in similar directions. Specifically, rain-making DNAs in Taipei Category 1 were correlated with wind force/direction and cloud height along PCs 1, 3, 4, and 7, and those in Category 2 were correlated with cloud-cover distribution along PCs 1, 2, and 5. The identified rain-making thresholds of typhoons with constant direction/structure showed a weaker steady state, whereas the unsteady rest produced multi-peak rainfall hydrographs. Rain evolution analysis reveals that cloudy rainbands, carried by the wind field, move along the Tamsui River valley when traveling between northeast and south-southeast of Taipei; converge with gradient and geostrophic winds when traveling between east-northeast and southwest; merge with southwest monsoon when traveling between west-southwest and northeast of Kaohsiung., Competing Interests: The authors declare the following financial interests/personal relationships which may be considered as potential competing interests:Chien-Lin Huang reports financial support was provided by 10.13039/100020595National Science and Technology Council, Taiwan., (© 2023 Published by Elsevier Ltd.)

Oxidized cysteine residues are highly reactive and can form functional covalent conjugates, of which the allosteric redox switch formed by the lysine-cysteine NOS bridge is an example. Here, we report a noncanonical FAD-dependent enzyme Orf1 that adds a glycine-derived N-formimidoyl group to glycinothricin to form the antibiotic BD-12. X-ray crystallography was used to investigate this complex enzymatic process, which showed Orf1 has two substrate-binding sites that sit 13.5 Å apart unlike canonical FAD-dependent oxidoreductases. One site could accommodate glycine and the other glycinothricin or glycylthricin. Moreover, an intermediate-enzyme adduct with a NOS-covalent linkage was observed in the later site, where it acts as a two-scissile-bond linkage facilitating nucleophilic addition and cofactor-free decarboxylation. The chain length of nucleophilic acceptors vies with bond cleavage sites at either N-O or O-S accounting for N-formimidoylation or N-iminoacetylation. The resultant product is no longer sensitive to aminoglycoside-modifying enzymes, a strategy that antibiotic-producing species employ to counter drug resistance in competing species., (© 2023. The Author(s).)

Targeted protein degradation has arisen as a powerful therapeutic modality for degrading disease targets. While proteolysis-targeting chimera (PROTAC) design is more modular, the discovery of molecular glue degraders has been more challenging. Here, we have coupled the phenotypic screening of a covalent ligand library with chemoproteomic approaches to rapidly discover a covalent molecular glue degrader and associated mechanisms. We have identified a cysteine-reactive covalent ligand EN450 that impairs leukemia cell viability in a NEDDylation and proteasome-dependent manner. Chemoproteomic profiling revealed covalent interaction of EN450 with an allosteric C111 in the E2 ubiquitin-conjugating enzyme UBE2D. Quantitative proteomic profiling revealed the degradation of the oncogenic transcription factor NFKB1 as a putative degradation target. Our study thus puts forth the discovery of a covalent molecular glue degrader that uniquely induced the proximity of an E2 with a transcription factor to induce its degradation in cancer cells., Competing Interests: Declaration of interests J.A.T., J.M.K., and D.D. are employees of Novartis Institutes for BioMedical Research. This study was funded by the Novartis Institutes for BioMedical Research and the Novartis-Berkeley Translational Chemical Biology Institute. D.K.N. is a co-founder, shareholder, and on the scientific advisory boards for Frontier Medicines and Vicinitas Therapeutics; a member of the board of directors for Vicinitas Therapeutics; on the scientific advisory boards of The Mark Foundation for Cancer Research, Photys Therapeutics, Apertor Pharmaceuticals, Ecto Therapeutics, Oerth Bio, and Chordia Therapeutics; and on the investment advisory board of Droia Ventures., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Glycidyl esters (GEs) and 2- and 3-monochloropropanediol esters (MCPDEs) are emerging process-generated food contaminants known as possible carcinogens. Herein, a direct method is developed and validated for the first time to simultaneously quantify seven GEs and twenty-four MCPDE congeners of processed foods using liquid chromatography-tandem mass spectrometry in a single sequence without ester cleavage or derivatisation, thereby allowing for the simultaneous analysis of numerous food matrices with high accuracy and precision. Our results show levels of GEs varying from

Tunicamycin (TUN) is a nucleoside antibiotic with a complex structure comprising uracil, tunicamine sugar, N -acetylglucosamine (GlcNAc), and fatty acyl tail moieties. TUN, known as a canonical inhibitor, blocks vital functions of certain transmembrane protein families, for example, the insect enzyme dolichyl phosphate α- N -acetylglucosaminylphosphotransferase (DPAGT1) of Spodoptera frugiperda and the bacterial enzyme phospho- N -acetylmuramoylpentapeptide translocase (MraY CB ) of Clostridium bolteae . Accurate description of protein-drug interactions has an immense impact on structure-based drug design, while the main challenge is to create proper topology and parameter entries for TUN in modeling protein-TUN interactions given the structural complexity. Starting from DPAGT1-TUN and MraY CB -TUN crystal structures, we first sketched these structural complexes on the basis of the CHARMM36 force field and optimized each of them using quantum mechanics/molecular mechanics (QM/MM) calculations. By continuing calculations on the active site (QM region) of each optimized structure, we specified the characteristics of intermolecular interactions contributing to the binding of TUN to each active site by quantum theory of atoms in molecules (QTAIM) and natural bond orbital (NBO) analyses at the M06-2X/6-31G** level. The results outlined that TUN insertion into each active site requires multiple weak, moderate, and strong hydrogen bonds accompanying charge-dipole, dipole-dipole, and hydrophobic interactions among different TUN moieties and adjacent residues. The water-mediated interactions also play central roles in situating the uracil and tunicamine moieties of TUN within the DPAGT1 active site as well as in preserving the uracil-binding pocket in the MraY CB active site. The TUN binds more strongly to DPAGT1 than to MraY CB . The information garnered here is valuable particularly for better understanding mode of action at the molecular level, as it is conducive to developing next generations of nucleoside antibiotics., Competing Interests: The authors declare no competing financial interest., (© 2022 The Authors. Published by American Chemical Society.)

This study aimed to evaluate the impact factors and effectiveness of management policies on the presence of polybrominated diphenyl ethers (PBDEs) in sediment samples in Taiwan from the last 10 years. Twenty-four PBDE congeners were detected in 838 sediment samples collected from 4 stages (2006-2019) in 30 principal rivers, based on the national project for background monitoring of the environmental distribution of chemical substances. The ΣPBDE concentrations in the 4 stages ranged from 30.00 to 147.10 ng/g dw, 6.03-15.30 ng/g dw, 4.99-7.00 ng/g dw, and 1.20-2.10 ng/g dw in the northern, southern, central, and eastern areas, respectively. The concentrations of PBDEs (e.g., penta-BDE and octa-BDE) in sediment samples notably decreased (-6 to -73%) as the Taiwan Environmental Protection Administration implemented policies banning PBDEs (except deca-BDE). The PBDEs levels of the sediment samples collected in the dry season were higher than those collected in the wet season. The levels of ΣPBDEs in sediment samples were affected by season, the amount of general waste present, and nearby PBDE-related factories and e-waste recycling facilities. Reducing the release of PBDEs, especially deca-BDE, through sound waste management and recycling practices is still needed to improve environmental sustainability in Taiwan., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Caprazamycin is a nucleoside antibiotic that inhibits phospho-N-acetylmuramyl-pentapeptide translocase (MraY). The biosynthesis of nucleoside antibiotics has been studied but is still far from completion. The present study characterized enzymes Cpz10, Cpz15, Cpz27, Mur17, Mur23 out of caprazamycin/muraymycin biosynthetic gene cluster, particularly the nonheme αKG-dependent enzyme Cpz10. Cpz15 is a β-hydroxylase converting uridine mono-phosphate to uridine 5' aldehyde, then incorporating with threonine by Mur17 (Cpz14) to form 5'-C-glycyluridine. Cpz10 hydroxylates synthetic 11 to 12 in vitro. Major product 13 derived from mutant Δcpz10 is phosphorylated by Cpz27. β-Hydroxylation of 11 by Cpz10 permits the maturation of caprazamycin, but decarboxylation of 11 by Mur23 oriented to muraymycin formation. Cpz10 recruits two iron atoms to activate dioxygen with regio-/stereo-specificity and commit electron/charge transfer, respectively. The chemo-physical interrogations should greatly advance our understanding of caprazamycin biosynthesis, which is conducive to pathway/protein engineering for developing more effective nucleoside antibiotics., (© 2022. The Author(s).)

COG-UK Mutation Explorer (COG-UK-ME, https://sars2.cvr.gla.ac.uk/cog-uk/-last accessed date 16 March 2022) is a web resource that displays knowledge and analyses on SARS-CoV-2 virus genome mutations and variants circulating in the UK, with a focus on the observed amino acid replacements that have an antigenic role in the context of the human humoral and cellular immune response. This analysis is based on more than 2 million genome sequences (as of March 2022) for UK SARS-CoV-2 data held in the CLIMB-COVID centralised data environment. COG-UK-ME curates these data and displays analyses that are cross-referenced to experimental data collated from the primary literature. The aim is to track mutations of immunological importance that are accumulating in current variants of concern and variants of interest that could alter the neutralising activity of monoclonal antibodies (mAbs), convalescent sera, and vaccines. Changes in epitopes recognised by T cells, including those where reduced T cell binding has been demonstrated, are reported. Mutations that have been shown to confer SARS-CoV-2 resistance to antiviral drugs are also included. Using visualisation tools, COG-UK-ME also allows users to identify the emergence of variants carrying mutations that could decrease the neutralising activity of both mAbs present in therapeutic cocktails, e.g. Ronapreve. COG-UK-ME tracks changes in the frequency of combinations of mutations and brings together the curated literature on the impact of those mutations on various functional aspects of the virus and therapeutics. Given the unpredictable nature of SARS-CoV-2 as exemplified by yet another variant of concern, Omicron, continued surveillance of SARS-CoV-2 remains imperative to monitor virus evolution linked to the efficacy of therapeutics., (© The Author(s) 2022. Published by Oxford University Press.)

Capreomycin (CMN) is an important second-line antituberculosis antibiotic isolated from Saccharothrix mutabilis subspecies capreolus . The gene cluster for CMN biosynthesis has been identified and sequenced, wherein the cph gene was annotated as a phosphotransferase likely engaging in self-resistance. Previous studies reported that Cph inactivates two CMNs, CMN IA and IIA, by phosphorylation. We, herein, report that (1) Escherichia coli harboring the cph gene becomes resistant to both CMN IIA and IIB, (2) phylogenetic analysis regroups Cph to a new clade in the phosphotransferase protein family, (3) Cph shares a three-dimensional structure akin to the aminoglycoside phosphotransferases with a high binding affinity ( K D ) to both CMN IIA and IIB at micromolar levels, and (4) Cph utilizes either ATP or GTP as a phosphate group donor transferring its γ-phosphate to the hydroxyl group of CMN IIA. Until now, Cph and Vph (viomycin phosphotransferase) are the only two known enzymes inactivating peptide-based antibiotics through phosphorylation. Our biochemical characterization and structural determination conclude that Cph confers the gene-carrying species resistance to CMN by means of either chemical modification or physical sequestration, a naturally manifested belt and braces strategy. These findings add a new chapter into the self-resistance of bioactive natural products, which is often overlooked while designing new bioactive molecules.

Kasugamycin (KSM), an aminoglycoside antibiotic, is composed of three chemical moieties: D- chiro -inositol, kasugamine and glycine imine. Despite being discovered more than 50 years ago, the biosynthetic pathway of KSM remains an unresolved puzzle. Here we report a structural and functional analysis for an epimerase, KasQ, that primes KSM biosynthesis rather than the previously proposed KasF/H, which instead acts as an acetyltransferase, inactivating KSM. Our biochemical and biophysical analysis determined that KasQ converts UDP-GlcNAc to UDP-ManNAc as the initial step in the biosynthetic pathway. The isotope-feeding study further confirmed that 13 C, 15 N-glucosamine/UDP-GlcNH 2 rather than glucose/UDP-Glc serves as the direct precursor for the formation of KSM. Both KasF and KasH were proposed, respectively, converting UDP-GlcNH 2 and KSM to UDP-GlcNAc and 2-N'-acetyl KSM. Experimentally, KasF is unable to do so; both KasF and KasH are instead KSM-modifying enzymes, while the latter is more specific and reactive than the former in terms of the extent of resistance. The information gained here lays the foundation for mapping out the complete KSM biosynthetic pathway.

We identify amino acid variants within dominant SARS-CoV-2 T cell epitopes by interrogating global sequence data. Several variants within nucleocapsid and ORF3a epitopes have arisen independently in multiple lineages and result in loss of recognition by epitope-specific T cells assessed by IFN-γ and cytotoxic killing assays. Complete loss of T cell responsiveness was seen due to Q213K in the A∗01:01-restricted CD8+ ORF3a epitope FTSDYYQLY 207-215 ; due to P13L, P13S, and P13T in the B∗27:05-restricted CD8+ nucleocapsid epitope QRNAPRITF 9-17 ; and due to T362I and P365S in the A∗03:01/A∗11:01-restricted CD8+ nucleocapsid epitope KTFPPTEPK 361-369 . CD8+ T cell lines unable to recognize variant epitopes have diverse T cell receptor repertoires. These data demonstrate the potential for T cell evasion and highlight the need for ongoing surveillance for variants capable of escaping T cell as well as humoral immunity., Competing Interests: The authors have no competing interests to declare., (© 2021 The Author(s).)

Background: In order to understand the molecular epidemiology of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) in Sri Lanka, since March 2020, we carried out genomic sequencing overlaid on available epidemiological data until April 2021. Methods: Whole genome sequencing was carried out on diagnostic sputum or nasopharyngeal swabs from 373 patients with COVID-19. Molecular clock phylogenetic analysis was undertaken to further explore dominant lineages. Results: The B.1.411 lineage was most prevalent, which was established in Sri Lanka and caused outbreaks throughout the country until March 2021. The estimated time of the most recent common ancestor (tMRCA) of this lineage was June 1, 2020 (with 95% lower and upper bounds March 30 to July 27) suggesting cryptic transmission may have occurred, prior to a large epidemic starting in October 2020. Returning travellers were identified with infections caused by lineage B.1.258, as well as the more transmissible B.1.1.7 lineage, which has replaced B.1.411 to fuel the ongoing large outbreak in the country. Conclusions: The large outbreak that started in early October, is due to spread of a single virus lineage, B.1.411 until the end of March 2021, when B.1.1.7 emerged and became the dominant lineage., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Jeewandara, Jayathilaka, Ranasinghe, Hsu, Ariyaratne, Jayadas, Panambara Arachchige, Lindsey, Gomes, Parker, Wijewickrama, Karunaratne, Ogg, de Silva and Malavige.)

We at the National Institute of Neurological Disorders and Stroke routinely receive questions and statements from trainees and faculty that suggest widespread beliefs about the necessity of a National Institutes of Health K99/R00 award, other prior funding, and/or specific types of publications for obtaining one's first tenure-track position in neuroscience. To address these beliefs, we examined the funding and publication history of a cohort of investigators who began their first academic faculty position between 2009 and 2019, and we interviewed several senior academic leaders with extensive experience in hiring new faculty. Our data show that <11% of newly hired faculty had a K99/R00 award and that neither prior funding nor papers in prestigious journals were necessary to obtain a tenure-track faculty position. Interviews with academic leaders almost uniformly referred to critically important factors that were considered to be more important in the hiring process than funding or publishing in high-profile journals., (Copyright © 2021 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC).)

In this article, we demonstrate that specifically engineered oxide nanoparticles (NPs) have the potential to act as theranostic materials that are able to generate or prevent oxidative stress through their oxi-redox activity in various types of malignant and nonmalignant cells. The oxi-redox activity is related to the type and presence of surface defects, which is modified with appropriate synthesis conditions. In the present work, we used MDA-MB-231 and MCF-7 human breast cancer cells and nonmalignant MCF-10A human breast cells to demonstrate how controlled oxidative stress mediated by specifically nanoengineered indium tin oxide (ITO) NPs can selectively induce cell death in the cancer cells while reducing the oxidative stress in the normal cells and supporting their proliferation. The ITO NPs are also promising nanotheranostic materials for cancer therapy and contrast agents because of their multimodal imaging capabilities. We demonstrate that the synthesized ITO NPs can selectively increase the generation of reactive oxygen species (ROS) in both breast tumor cell lines, resulting in activation of apoptosis, and can also greatly suppress the cellular proliferation in both types of tumor cells. In contrast, the ITO NPs exhibit ROS scavenging-like behavior, significantly decreasing the ROS levels in MCF-10A cells exposed to the additional ROS, hydrogen peroxide (H 2 O 2 ), so that they protect the proliferation of nonmalignant MCF-10A cells from ROS damage. In addition, fluorescent microscopy images revealed that the ITO NPs emit strong fluorescence that could be used to reveal their location. Moreover, computed tomography imaging demonstrated that the ITO NPs exhibited a comparable capability toward anatomical contrast enhancement. These results suggest that the synthesized ITO NPs have the potential to be a novel selective therapeutic agent with a multimodal imaging property for anticancer treatment.

Incremental language processing means that listeners confront temporary ambiguity about how to structure the input, which can generate misinterpretations. In four "visual-world" experiments, we tested whether engaging cognitive control - which detects and resolves conflict - assists revision during comprehension. We recorded listeners' eye-movements and actions while following instructions that were ripe for misanalysis. In Experiments 1 and 3, sentences followed trials from a nonverbal conflict task that manipulated cognitive-control engagement, to test its impact on the ability to revise. To isolate conflict-driven effects of cognitive-control on comprehension, we manipulated attention in a non-conflict task in Experiments 2 and 4. We observed fewer comprehension errors, and earlier revision, when cognitive control (more than attention) was elicited on an immediately preceding trial. These results extend previous correlations between cognitive control and language processing by revealing the influence of domain-general cognitive-control engagement on the temporal unfolding of error-revision processes during language comprehension., Competing Interests: Disclosure statement No potential conflict of interest was reported by the author(s).

New neurotechnologies fueled by the BRAIN Initiative now allow investigators to map, monitor and modulate complex neural circuits, enabling the pursuit of research questions previously considered unapproachable. Yet it is the convergence of molecular neuroscience with the new systems neuroscience that promises the greatest future advances. This is particularly true for our understanding of nervous system disorders, some of which have known molecular drivers or pathology but result in unknown perturbations in circuit function. NIH-supported research on "BRAIN Circuits" programs integrate experimental, analytic, and theoretical capabilities for analysis of specific neural circuits and their contributions to perceptions, motivations, and actions. In this review, we describe the BRAIN priority areas, review our strategy for balancing early feasibility with mature projects, and the balance of individual with team science for this 'BRAIN Circuits' program. We also highlight the diverse portfolio of techniques, species, and neural systems represented in these projects., (Published by Elsevier Ltd.)

Two systems of antibody-drug conjugates (ADCs), noncleavable H32-DM1 and cleavable H32-VCMMAE, were developed by using different linkers and drugs attached to the anti-HER2 antibody H32, which is capable of cell internalization. Activated functional groups, including an N-hydroxysuccinimidyl (NHS) ester and a maleimide, were utilized to make the ADCs. Mass spectrometry, hydrophobic interaction chromatography, polyacrylamide gel electrophoresis, and in vitro cell assays were performed to analyze and optimize the ADCs. Several H32-VCMMAE ADCs were established with higher DARs and greater synthetic yields without compromising potency. The anticancer efficacy of H32-DM1 was 2- to 8-fold greater than that of Kadcyla®. The efficacy of H32-VCMMAE was in turn better than that of H32-DM1. The anticancer efficacy of these ADCs against N87, SK-BR-3 and BT474 cells was in the following order: H32-VCMMAE series > H32-DM1 series > Kadcyla®. The optimal DAR for H32-VCMMAE was found to be 6.6, with desirable attributes including good cell penetration, a releasable payload in cancer cells, and high potency. Our results demonstrated the potential of H32-VCMMAE as a good ADC candidate., Competing Interests: The authors have declared that no competing interests exist.

Linker design is crucial to the success of antibody-drug conjugates (ADCs). In this work, we developed a modular linker format for attaching molecular cargos to antibodies based on strand pairing between complementary oligonucleotides. We prepared antibody-oligonucleotide conjugates (AOCs) by attaching 18-mer oligonucleotides to an anti-HER2 antibody through thiol-maleimide chemistry, a method generally applicable to any immunoglobulin with interchain disulfide bridges. The hybridization of drug-bearing complementary oligonucleotides to our AOCs was rapid, stoichiometric, and sequence-specific. AOCs loaded with cytotoxic payloads were able to selectively target HER2-overexpressing cell lines such as SK-BR-3 and N87, with in vitro potencies similar to that of the marketed ADC Kadcyla (T-DM1). Our results demonstrated the potential of utilizing AOCs as a highly versatile and modular platform, where a panel of well-characterized AOCs bearing DNA, RNA, or various nucleic acid analogs, such as peptide nucleic acids, could be easily paired with any cargo of choice for a wide range of diagnostic or therapeutic applications.

Though reactive flavin-N5/C4α-oxide intermediates can be spectroscopically profiled for some flavin-assisted enzymatic reactions, their exact chemical configurations are hardly visualized. Structural systems biology and stable isotopic labelling techniques were exploited to correct this stereotypical view. Three transition-like complexes, the α-ketoacid…N5-FMN ox complex (I), the FMN ox -N5-aloxyl-C'α - -C4α + zwitterion (II), and the FMN-N5-ethenol-N5-C4α-epoxide (III), were determined from mandelate oxidase (Hmo) or its mutant Y128F (monooxygenase) crystals soaked with monofluoropyruvate (a product mimic), establishing that N5 of FMN ox an alternative reaction center can polarize to an ylide-like mesomer in the active site. In contrast, four distinct flavin-C4α-oxide adducts (IV-VII) from Y128F crystals soaked with selected substrates materialize C4α of FMN an intrinsic reaction center, witnessing oxidation, Baeyer-Villiger/peroxide-assisted decarboxylation, and epoxidation reactions. In conjunction with stopped-flow kinetics, the multifaceted flavin-dependent reaction continuum is physically dissected at molecular level for the first time., (© 2020 The Authors. Protein Science published by Wiley Periodicals, Inc. on behalf of The Protein Society.)

The Y128F single mutant of p-hydroxymandelate oxidase (Hmo) is capable of oxidizing mandelate to benzoate via a four-electron oxidative decarboxylation reaction. When benzoylformate (the product of the first two-electron oxidation) and hydrogen peroxide (an oxidant) were used as substrates the reaction did not proceed, suggesting that free hydrogen peroxide is not the committed oxidant in the second two-electron oxidation. How the flavin mononucleotide (FMN)-dependent four-electron oxidation reaction takes place remains elusive. Structural and biochemical explorations have shed new light on this issue. 15 high-resolution crystal structures of Hmo and its mutants liganded with or without a substrate reveal that oxidized FMN (FMN ox ) possesses a previously unknown electrophilic/nucleophilic duality. In the Y128F mutant the active-site perturbation ensemble facilitates the polarization of FMN ox to a nucleophilic ylide, which is in a position to act on an α-ketoacid, forming an N5-acyl-FMN red dead-end adduct. In four-electron oxidation, an intramolecular disproportionation reaction via an N5-alkanol-FMN red C'α carbanion intermediate may account for the ThDP/PLP/NADPH-independent oxidative decarboxylation reaction. A synthetic 5-deaza-FMN ox cofactor in combination with an α-hydroxyamide or α-ketoamide biochemically and structurally supports the proposed mechanism., (open access.)

p-Hydroxymandelate oxidase (Hmo) is a flavin mononucleotide (FMN)-dependent enzyme that oxidizes mandelate to benzoylformate. How the FMN-dependent oxidation is executed by Hmo remains unclear at the molecular level. A continuum of snapshots from crystal structures of Hmo and its mutants in complex with physiological/nonphysiological substrates, products and inhibitors provides a rationale for its substrate enantioselectivity/promiscuity, its active-site geometry/reactivity and its direct hydride-transfer mechanism. A single mutant, Y128F, that extends the two-electron oxidation reaction to a four-electron oxidative decarboxylation reaction was unexpectedly observed. Biochemical and structural approaches, including biochemistry, kinetics, stable isotope labeling and X-ray crystallography, were exploited to reach these conclusions and provide additional insights., (open access.)

Lipoglycopeptide antibiotics, for example, teicoplanin (Tei) and A40926, are more potent than vancomycin against Gram-positive (Gram-(+)) drug-resistant pathogens, for example, methicillin-resistant Staphylococcus aureus (MRSA). To extend their therapeutic effectiveness on vancomycin-resistant S. aureus (VRSA), the biosynthetic pathway of the N-acyl glucosamine (Glc) pharmacophore at residue 4 (r4) of teicoplanin pseudoaglycone redirection to residue 6 (r6) was attempted. On the basis of crystal structures, two regioselective biocatalysts Orf2*T (a triple-mutation mutant S98A/V121A/F193Y) and Orf11*S (a single-mutation mutant W163A) were engineered, allowing them to act on GlcNAc at r6. New analogs thereby made show marked antimicrobial activity against MRSA and VRSA by 2-3 orders of magnitude better than teicoplanin and vancomycin. The lipid side chain of the Tei-analogs armed with a terminal mono- or diguanidino group extends the antimicrobial specificity from Gram-(+) to Gram-negative (Gram-(-)), comparable to that of kanamycin. In addition to low cytotoxicity and high safety, the Tei analogs exhibit new modes of action as a result of resensitization of VRSA and Acinetobacter baumannii. The redirection of the biosynthetic pathway for the N-acyl-Glc pharmacophore from r4 to r6 bodes well for large-scale production of selected r6,Tei congeners in an environmentally friendly synthetic biology approach.

Transketolase (TK) catalyzes a reversible transfer of a two-carbon (C 2 ) unit between phosphoketose donors and phosphoaldose acceptors, for which the group-transfer reaction that follows a one- or two-electron mechanism and the force that breaks the C2"-C3" bond of the ketose donors remain unresolved. Herein, we report ultrahigh-resolution crystal structures of a TK (TKps) from Pichia stipitis in previously undiscovered intermediate states and support a diradical mechanism for a reversible group-transfer reaction. In conjunction with MS, NMR spectroscopy, EPR and computational analyses, it is concluded that the enzyme-catalyzed non-Kekulé diradical cofactor brings about the C2"-C3" bond cleavage/formation for the C 2 -unit transfer reaction, for which suppression of activation energy and activation and destabilization of enzymatic intermediates are facilitated., (© 2018 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA.)

Thematic role assignment - generally, figuring out who did what to whom - is a critical component of sentence comprehension, which is influenced by both syntactic and semantic cues. Conflict between these cues can result in temporary consideration of multiple incompatible interpretations during real-time sentence processing. We tested whether the resolution of syntax-semantics conflict can be expedited by the online engagement of cognitive control processes that are routinely used to regulate behavior across domains. In this study, cognitive control deployment from a previous Stroop trial influenced eye movements during subsequent sentence comprehension. Specifically, when syntactic and semantic cues competed for influence on interpretation, dynamic cognitive control engagement led to (a) fewer overall looks to a picture illustrating the competing but incorrect interpretation (Experiment 1), or (b) steeper growth in looks to a picture illustrating the correct interpretation (Experiment 2). Thus, prior cognitive control engagement facilitated the resolution of syntax-semantics conflict by biasing processing towards the intended analysis. This conflict adaptation effect demonstrates a causal connection between cognitive control and real-time thematic role assignment. Broader patterns demonstrated that prior cognitive control engagement also modulated sentence processing irrespective of the presence of conflict, reflecting increased integration of newly arriving cues with prior sentential content. Together, the results suggest that cognitive control helps listeners determine correct event roles during real-time comprehension., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The BRAIN Initiative arose from a grand challenge to "accelerate the development and application of new technologies that will enable researchers to produce dynamic pictures of the brain that show how individual brain cells and complex neural circuits interact at the speed of thought." The BRAIN Initiative is a public-private effort focused on the development and use of powerful tools for acquiring fundamental insights about how information processing occurs in the central nervous system (CNS). As the Initiative enters its fifth year, NIH has supported >500 principal investigators, who have answered the Initiative's challenge via hundreds of publications describing novel tools, methods, and discoveries that address the Initiative's seven scientific priorities. We describe scientific advances produced by individual laboratories, multi-investigator teams, and entire consortia that, over the coming decades, will produce more comprehensive and dynamic maps of the brain, deepen our understanding of how circuit activity can produce a rich tapestry of behaviors, and lay the foundation for understanding how its circuitry is disrupted in brain disorders. Much more work remains to bring this vision to fruition, and the National Institutes of Health continues to look to the diverse scientific community, from mathematics, to physics, chemistry, engineering, neuroethics, and neuroscience, to ensure that the greatest scientific benefit arises from this unique research Initiative., (Copyright © 2018 the authors 0270-6474/18/386427-12$15.00/0.)

It is theoretically plausible that thiazolium mesomerizes to congeners other than carbene in a low effective dielectric binding site; especially given the energetics and uneven electronegativity of carbene groups. However, such a phenomenon has never been reported. Nine crystal structures of transketolase obtained from Pichia stipitis (TKps) are reported with subatomic resolution, where thiazolium displays an extraordinary ring-bending effect. The bent thiazolium congeners correlate with non-Kekulé diradicals because there is no gain or loss of electrons. In conjunction with biophysical and biochemical analyses, it is concluded that ring bending is a result of tautomerization of thiazolium with its non- Kekulé diradicals, exclusively in the binding site of TKps. The chemophysical properties of these thiazolium mesomers may account for the great variety of reactivities carried out by thiamine-diphosphate-containing (ThDP) enzymes. The stability of ThDP in living systems can be regulated by the levels of substrates, and hydration and dehydration, as well as diradical-mediated oxidative degradation., (© 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Regions within the left inferior frontal gyrus (LIFG) have simultaneously been implicated in syntactic processing and cognitive control. Accounts attempting to unify LIFG's function hypothesize that, during comprehension, cognitive control resolves conflict between incompatible representations of sentence meaning. Some studies demonstrate co-localized activity within LIFG for syntactic and non-syntactic conflict resolution, suggesting domain-generality, but others show non-overlapping activity, suggesting domain-specific cognitive control and/or regions that respond uniquely to syntax. We propose however that examining exclusive activation sites for certain contrasts creates a false dichotomy: both domain-general and domain-specific neural machinery must coordinate to facilitate conflict resolution across domains. Here, subjects completed four diverse tasks involving conflict -one syntactic, three non-syntactic- while undergoing fMRI. Though LIFG consistently activated within individuals during conflict processing, functional connectivity analyses revealed task-specific coordination with distinct brain networks. Thus, LIFG may function as a conflict-resolution "hub" that cooperates with specialized neural systems according to information content., (Copyright © 2016 Elsevier Inc. All rights reserved.)

In the development of new functionalities of transketolase for the industrial strain Pichia stipitis (TKps) the structural information of TKps would allow us to gain insight into the enzyme's reaction mechanisms, substrates selectivity and reaction directionality to help reach the goal. We here report seven TKps crystal structures of wild type (WT) and mutants in complex with various physiological ligands. These complexes were refined to resolutions at 1.6-1.03 Å. Both biochemical and mutagenic analyses concluded that residues His27, His66, His100, His261, His478, Asp473, Arg356 and Arg525 play important roles in coenzyme binding and substrates recognition. In general, His66 and His261 hold thiamine diphosphate in place; Arg356 and Arg525 serve as gatekeepers interacting with the terminal phosphate group of sugar-phosphates. His27, His66, His100, His478 and Asp473 are critical for sugars recognition/binding, in which His27 is relatively more important in interaction with sedoheptulose-7-phosphate (S7P) than xylulose-5-phosphate (X5P) in terms of molecular recognition/binding affinity. Kinetically, the reactions with X5P (forward) which were catalyzed by WT or H27A are indistinguishable, while in the reactions with S7P (backward) H27A exhibits weaker activity relative to WT. As a result, given TKps(H27A) as the biocatalyst the overall reactivity reverses from the backward reaction preference to forward, thus facilitating net xylose assimilation., (© The Author 2016. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.)

Siderophores, such as enterobactin (Ent), are small molecules that can be selectively imported into bacteria along with iron by cognate transporters. Siderophore conjugates are thus a promising strategy for delivering functional reagents into bacteria. In this work, we present an easy-to-perform, one-pot chemoenzymatic synthesis of functionalized monoglucosylated enterobactin (MGE). When functionalized MGE is conjugated to a rhodamine fluorophore, which affords RhB-Glc-Ent, it can selectively label Gram-negative bacteria that utilize Ent, including some E. coli strains and P. aeruginosa. V. cholerae, a bacterium that utilizes linearized Ent, can also be weakly targeted. Moreover, the targeting is effective under iron-limiting but not iron-rich conditions. Our results suggest that the RhB-Glc-Ent probe is sensitive not only to the bacterial strain but also to the iron condition in the environment., (© 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Speech unfolds swiftly, yet listeners keep pace by rapidly assigning meaning to what they hear. Sometimes, though, initial interpretations turn out to be wrong. How do listeners revise misinterpretations of language input moment by moment to avoid comprehension errors? Cognitive control may play a role by detecting when processing has gone awry and then initiating behavioral adjustments accordingly. However, no research to date has investigated a cause-and-effect interplay between cognitive-control engagement and the overriding of erroneous interpretations in real time. Using a novel cross-task paradigm, we showed that Stroop-conflict detection, which mobilizes cognitive-control procedures, subsequently facilitates listeners' incremental processing of temporarily ambiguous spoken instructions that induce brief misinterpretation. When instructions followed incongruent Stroop items, compared with congruent Stroop items, listeners' eye movements to objects in a scene reflected more transient consideration of the false interpretation and earlier recovery of the correct one. Comprehension errors also decreased. Cognitive-control engagement therefore accelerates sentence-reinterpretation processes, even as linguistic input is still unfolding., (© The Author(s) 2016.)

Many features can describe a concept, but only some features define a concept in that they enable discrimination of items that are instances of a concept from (similar) items that are not. We refer to this property of some features as feature diagnosticity. Previous work has described the behavioral effects of feature diagnosticity, but there has been little work on explaining why and how these effects arise. In this study, we aimed to understand the impact of feature diagnosticity on concept representations across two complementary experiments. In Experiment 1, we manipulated the diagnosticity of one feature, color, for a set of novel objects that human participants learned over the course of 1 week. We report behavioral and neural evidence that diagnostic features are likely to be automatically recruited during remembering. Specifically, individuals activated color-selective regions of ventral temporal cortex (specifically, left fusiform gyrus and left inferior temporal gyrus) when thinking about the novel objects, although color information was never explicitly probed during the task. Moreover, multiple behavioral and neural measures of the effects of feature diagnosticity were correlated across participants. In Experiment 2, we examined relative color association in familiar object categories, which varied in feature diagnosticity (fruits and vegetables, household items). Taken together, these results offer novel insights into the neural mechanisms underlying concept representations by demonstrating that automatic recruitment of diagnostic information gives rise to behavioral effects of feature diagnosticity.

Teicoplanin A2-2 (Tei)/A40926 is the last-line antibiotic to treat multidrug-resistant Gram-positive bacterial infections, e.g., methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant enterococcus (VRE). This class of antibiotics is powered by the N-acyltransferase (NAT) Orf11*/Dbv8 through N-acylation on glucosamine at the central residue of Tei/A40926 pseudoaglycone. The NAT enzyme possesses enormous value in untapped applications; its advanced development is hampered largely due to a lack of structural information. In this report, we present eight high-resolution X-ray crystallographic unary, binary, and ternary complexes in order to decipher the molecular basis for NAT's functionality. The enzyme undergoes a multistage conformational change upon binding of acyl-CoA, thus allowing the uploading of Tei pseudoaglycone to enable the acyl-transfer reaction to take place in the occlusion between the N- and C-halves of the protein. The acyl moiety of acyl-CoA can be bulky or lengthy, allowing a large extent of diversity in new derivatives that can be formed upon its transfer. Vancomycin/synthetic acyl-N-acetyl cysteamine was not expected to be able to serve as a surrogate for an acyl acceptor/donor, respectively. Most strikingly, NAT can catalyze formation of 2-N,6-O-diacylated or C6→C2 acyl-substituted Tei analogues through an unusual 1,4-migration mechanism under stoichiometric/solvational reaction control, wherein selected representatives showed excellent biological activities, effectively counteracting major types (VanABC) of VRE.

The purpose of this study is to establish a turbidity forecasting model as well as an early-warning system for turbidity management using rainfall records as the input variables. The Taipei Water Source Domain was employed as the study area, and ANOVA analysis showed that the accumulative rainfall records of 1-day Ping-lin, 2-day Ping-lin, 2-day Fei-tsui, 2-day Shi-san-gu, 2-day Tai-pin and 2-day Tong-hou were the six most significant parameters for downstream turbidity development. The artificial neural network model was developed and proven capable of predicting the turbidity concentration in the investigated catchment downstream area. The observed and model-calculated turbidity data were applied to developing the turbidity early-warning system. Using a previously determined turbidity as the threshold, the rainfall criterion, above which the downstream turbidity would possibly exceed this respective threshold turbidity, for the investigated rain gauge stations was determined. An exemplary illustration demonstrated the effectiveness of the proposed turbidity early-warning system as a precautionary alarm of possible significant increase of downstream turbidity. This study is the first report of the establishment of the turbidity early-warning system. Hopefully, this system can be applied to source water turbidity forecasting during storm events and provide a useful reference for subsequent adjustment of drinking water treatment operation.

Executive control (EC) generally refers to the regulation of mental activity. It plays a crucial role in complex cognition, and EC skills predict high-level abilities including language processing, memory, and problem solving, as well as practically relevant outcomes such as scholastic achievement. EC develops relatively late in ontogeny, and many sub-groups of developmental populations demonstrate an exaggeratedly poor ability to control cognition even alongside the normal protracted growth of EC skills. Given the value of EC to human performance, researchers have sought means to improve it through targeted training; indeed, accumulating evidence suggests that regulatory processes are malleable through experience and practice. Nonetheless, there is a need to understand both whether specific populations might particularly benefit from training, and what cortical mechanisms engage during performance of the tasks used in the training protocols. This contribution has two parts: in Part I, we review EC development and intervention work in select populations. Although promising, the mixed results in this early field make it difficult to draw strong conclusions. To guide future studies, in Part II, we discuss training studies that have included a neuroimaging component - a relatively new enterprise that also has not yet yielded a consistent pattern of results post-training, preventing broad conclusions. We therefore suggest that recent developments in neuroimaging (e.g., multivariate and connectivity approaches) may be useful to advance our understanding of the neural mechanisms underlying the malleability of EC and brain plasticity. In conjunction with behavioral data, these methods may further inform our understanding of the brain-behavior relationship and the extent to which EC is dynamic and malleable, guiding the development of future, targeted interventions to promote executive functioning in both healthy and atypical populations.

In biological systems, methylation is most commonly performed by methyltransferases (MTs) using the electrophilic methyl source S-adenosyl-L-methionine (SAM) via the S(N)2 mechanism. (2S,3S)-β-Methylphenylalanine, a nonproteinogenic amino acid, is a building unit of the glycopeptide antibiotic mannopeptimycin. The gene product of mppJ from the mannopeptimycin-biosynthetic gene cluster is the MT that methylates the benzylic C atom of phenylpyruvate (Ppy) to give βMePpy. Although the benzylic C atom of Ppy is acidic, how its nucleophilicity is further enhanced to become an acceptor for C-methylation has not conclusively been determined. Here, a structural approach is used to address the mechanism of MppJ and to engineer it for new functions. The purified MppJ displays a turquoise colour, implying the presence of a metal ion. The crystal structures reveal MppJ to be the first ferric ion SAM-dependent MT. An additional four structures of binary and ternary complexes illustrate the molecular mechanism for the metal ion-dependent methyltransfer reaction. Overall, MppJ has a nonhaem iron centre that bind, orients and activates the α-ketoacid substrate and has developed a sandwiched bi-water device to avoid the formation of the unwanted reactive oxo-iron(IV) species during the C-methylation reaction. This discovery further prompted the conversion of the MT into a structurally/functionally unrelated new enzyme. Through stepwise mutagenesis and manipulation of coordination chemistry, MppJ was engineered to perform both Lewis acid-assisted hydration and/or O-methyltransfer reactions to give stereospecific new compounds. This process was validated by six crystal structures. The results reported in this study will facilitate the development and design of new biocatalysts for difficult-to-synthesize biochemicals.

Optimization methods usually obtain the travel direction of the solution by substituting the solutions into the objective function. However, if the solution space is too large, this search method may be time consuming. In order to address this problem, this study incorporated the Taguchi method into the solution space search process of the optimization method, and used the characteristics of the Taguchi method to sequence the effects of the variation of decision variables on the system. Based on the level of effect, this study determined the impact factor of decision variables and the optimal solution for the model. The integration of the Taguchi method and the solution optimization method successfully obtained the optimal solution of the optimization problem, while significantly reducing the solution computing time and enhancing the river water quality. The results suggested that the basin with the greatest water quality improvement effectiveness is the Dahan River. Under the optimal strategy of this study, the severe pollution length was reduced from 18 km to 5 km.

Streptothricin-F (STT-F), one of the early-discovered antibiotics, consists of three components, a β-lysine homopolymer, an aminosugar D-gulosamine, and an unusual bicyclic streptolidine. The biosynthesis of streptolidine is a long-lasting but unresolved puzzle. Herein, a combination of genetic/biochemical/structural approaches was used to unravel this problem. The STT gene cluster was first sequenced from a Streptomyces variant BCRC 12163, wherein two gene products OrfP and OrfR were characterized in vitro to be a dihydroxylase and a cyclase, respectively. Thirteen high-resolution crystal structures for both enzymes in different reaction intermediate states were snapshotted to help elucidate their catalytic mechanisms. OrfP catalyzes an Fe(II) -dependent double hydroxylation reaction converting L-Arg into (3R,4R)-(OH)2 -L-Arg via (3S)-OH-L-Arg, while OrfR catalyzes an unusual PLP-dependent elimination/addition reaction cyclizing (3R,4R)-(OH)2 -L-Arg to the six-membered (4R)-OH-capreomycidine. The biosynthetic mystery finally comes to light as the latter product was incorporation into STT-F by a feeding experiment., (Copyright © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Cognitive control, otherwise known as executive function, refers to our ability to flexibly adjust or regulate habitual actions or behaviors. As a cluster of separable components, it depends heavily on the prefrontal cortex, one of the last brain regions to reach adult maturity. Cognitive control processes are thought to be among the key factors for scholastic success, and thus, underdeveloped cognitive control abilities might contribute to an achievement gap. In this chapter, we first discuss the prolonged maturation of the prefrontal cortex that leads to delayed emergence of cognitive control abilities in children. We briefly describe some of the functional effects of prolonged maturation of the prefrontal cortex. We then discuss how experience and environmental factors such as education and socioeconomic status may affect the development of cognitive control abilities, before turning to cognitive training interventions as a promising avenue for reducing this cognitive "gap" in both healthy children and those with developmental disabilities. Taken together, our hope is that by understanding the interaction of brain development, environmental factors, and the promise of cognitive interventions in children, this knowledge can help to both guide educational achievement and inform educational policy.

Ionic liquid pretreatment of biomass has been shown to greatly reduce the recalcitrance of lignocellulosic biomass, resulting in improved sugar yields after enzymatic saccharification. However, even under these improved saccharification conditions the cost of enzymes still represents a significant proportion of the total cost of producing sugars and ultimately fuels from lignocellulosic biomass. Much of the high cost of enzymes is due to the low catalytic efficiency and stability of lignocellulolytic enzymes, especially cellulases, under conditions that include high temperatures and the presence of residual pretreatment chemicals, such as acids, organic solvents, bases, or ionic liquids. Improving the efficiency of the saccharification process on ionic liquid pretreated biomass will facilitate reduced enzyme loading and cost. Thermophilic cellulases have been shown to be stable and active in ionic liquids but their activity is typically at lower levels. Cel5A&#95;Tma, a thermophilic endoglucanase from Thermotoga maritima, is highly active on cellulosic substrates and is stable in ionic liquid environments. Here, our motivation was to engineer mutants of Cel5A&#95;Tma with higher activity on 1-ethyl-3-methylimidazolium acetate ([C2mim][OAc]) pretreated biomass. We developed a robotic platform to screen a random mutagenesis library of Cel5A&#95;Tma. Twelve mutants with 25-42% improvement in specific activity on carboxymethyl cellulose and up to 30% improvement on ionic-liquid pretreated switchgrass were successfully isolated and characterized from a library of twenty thousand variants. Interestingly, most of the mutations in the improved variants are located distally to the active site on the protein surface and are not directly involved with substrate binding.

Discharge measurement in a tidal stream is always a difficult task. Owing to the complex flow conditions, discharge measurement in tidal streams has to be finished quickly, and must be highly efficient in order to yield an accurate measurement of real-time discharge. Measuring the discharge of tidal streams is done in three steps: (1) establishing the stage and cross-sectional area relation, (2) estimating the index velocity by using a velocity distribution equation based on the probability concept, (3) establishing the relationship between the index and mean velocities. Then the cross-sectional area and mean velocity can be estimated by the gage height and index velocity, respectively. The discharge of tidal streams is computed as the product of the cross-sectional area and mean velocity. The velocity distribution of the Taipei Bridge and Guan-du Bridge in the Tanshui River were established and analyzed to demonstrate the use of discharge measurement by index velocity for estimating the discharge of this tidal stream. The results reveal no large difference between the discharges measured by the conventional methods and the index velocity. This pilot study proposed a cost-effectiveness and efficient method. It is an easy, quick, and accurate model for measuring the real-time discharge of a tidal stream. It makes automatic, real-time, and continuous monitoring of the discharge in a tidal stream become possible.

In our effort to improve the efficiency and yield of xylose-to-ethanol bioconversion in Pichia stipitis, the transaldolase (TAL) in the pentose phosphate pathway was identified as a rate-limiting enzyme for improvement. A mutant containing the Q263R change was first obtained by directed evolution with 5-fold increase of activity, which was then incorporated into P. stipitesvia the pYDS vector to produce a genetically stable strain for fermentation on xylose. In comparison with the parental strain, TAL-Q263R(+) increases ethanol prodcution by 36% and 100% as measured by volumetric production rate and specific production rate, respectively. Thus improving the transaldolase activity in P. stipitis can significantly increase the rate and yield of xylose conversion to ethanol.

Sensorimotor theories of semantic memory require overlap between conceptual and perceptual representations. One source of evidence for such overlap comes from neuroimaging reports of co-activation during memory retrieval and perception; for example, regions involved in color perception (i.e., regions that respond more to colored than grayscale stimuli) are activated by retrieval of object color. One unanswered question from these studies is whether distinctions that are observed during perception are likewise observed during memory retrieval. That is, are regions defined by a chromaticity effect in perception similarly modulated by the chromaticity of remembered objects (e.g., lemons more than coal)? Subjects performed color perception and color retrieval tasks while undergoing fMRI. We observed increased activation during both perception and memory retrieval of chromatic compared to achromatic stimuli in overlapping areas of the left lingual gyrus, but not in dorsal or anterior regions activated during color perception. These results support sensorimotor theories but suggest important distinctions within the conceptual system., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

Neuroimaging tests of sensorimotor theories of semantic memory hinge on the extent to which similar activation patterns are observed during perception and retrieval of objects or object properties. The present study was motivated by the hypothesis that some of the seeming discrepancies across studies reflect flexibility in the systems responsible for conceptual and perceptual processing of color. Specifically, we test the hypothesis that retrieval of color knowledge can be influenced by both context (a task variable) and individual differences in cognitive style (a subject variable). In Experiment 1, we provide fMRI evidence for differential activity during color knowledge retrieval by having subjects perform a verbal task, in which context encouraged subjects to retrieve more- or less-detailed information about the colors of named common objects in a blocked experimental design. In the left fusiform, we found more activity during retrieval of more- versus less-detailed color knowledge. We also assessed preference for verbal or visual cognitive style, finding that brain activity in the left lingual gyrus significantly correlated with preference for a visual cognitive style. We replicated many of these effects in Experiment 2, in which stimuli were presented more quickly, in a random order, and in the auditory modality. This illustration of some of the factors that can influence color knowledge retrieval leads to the conclusion that tests of conceptual and perceptual overlap must consider variation in both of these processes.

Mannopeptimycin, a potent drug lead, has superior activity against difficult-to-treat multidrug-resistant Gram-positive pathogens such as methicillin-resistant Staphylococcus aureus (MRSA). (2S,3S)-beta-Methylphenylalanine is a residue in the cyclic hexapeptide core of mannopeptimycin, but the synthesis of this residue is far from clear. We report here on the reaction order and the stereochemical course of reaction in the formation of (2S,3S)-beta-methylphenylalanine. The reaction is executed by the enzymes MppJ and TyrB, an S-adenosyl methionine (SAM)-dependent methyltransferase and an (S)-aromatic-amino-acid aminotransferase, respectively. Phenylpyruvic acid is methylated by MppJ at its benzylic position at the expense of one equivalent of SAM. The resulting beta-methyl phenylpyruvic acid is then converted to (2S,3S)-beta-methylphenylalanine by TyrB. MppJ was further determined to be regioselective and stereoselective in its catalysis of the formation of (3S)-beta-methylphenylpyruvic acid. The binding constant (K(D)) of MppJ versus SAM is 26 microM. The kinetic constants with respect to k(cat Ppy) and K(M Ppy), and k(cat SAM) and K(M SAM) are 0.8 s(-1) and 2.5 mM, and 8.15 s(-1) and 0.014 mM, respectively. These results suggest SAM has higher binding affinity for MppJ than Ppy, and the C--C bond formation in betamPpy might be the rate-limiting step, as opposed to the C--S bond breakage in SAM.

A total of 46 carbapenem- and multidrug-resistant (CR- and MDR-)Acinetobacter baumannii bacteremic isolates from a Taiwanese medical center were investigated over the period 2000 to 2006 using randomly amplified polymorphic DNA (RAPD) profiling and by analysing the genetic organization of their integrons. The results of RAPD patterns revealed that before 2003 each CR- and MDR-A. baumannii bacteremic isolate was independent, but after 2003 the isolates appeared to belong in four epidemic strains and persisted in the hospital. All the CR- and MDR-A. baumannii strains harbored class I integron (intI1) genes. PCR amplification and nucleotide sequencing showed that the cassette genes of intI1 were found to form four different antibiotic-resistant gene alignments in those strains. The bla(IMP-1) gene in the cassette genes of intI1 was identified in a clone, which raised great concern that clonal spread of this strain or of an integron-mediated horizontal gene may have occurred.