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In this study, surface-enhanced Raman scattering substrates were investigated by the electrodeposition method to detect low concentrations of pesticides via the electrodeposition method with different agents from silver and gold precursors on APTES-modified ITO glass. A dual-potential method supplied three electrodes and was performed with a nucleation potential of 0.7 V for 2 s and a growth potential of -0.2 V for 500 s. The Ag film produced by the electrodeposition approach has great surface uniformity and good SERS signal amplification for the thiram insecticide at low concentrations. Interestingly, the ITO/APTES/Ag substrate extensively increased the sensitivity than the other investigated ones, thanks to the adequate assistance of amino groups of APTES in the denser and hierarchical deposition of Ag NPs. These observations were additionally elucidated via finite-difference time-domain (FDTD) calculation. For thiram, the detection was set at 10-8 M with an enhancement factor of up to 3.6 × 107 times. Comparing the SERS spectra of thiram at concentrations of 10-3, 10-4, and 10-5 M with a relative standard deviation (RSD) of less than 7.0% demonstrates excellent reproducibility of the ITO/APTES/Ag substrate. In addition, the special selectivity of the ITO/APTES/Ag substrate for thiram demonstrates that these nanostructures can identify pesticides with extreme sensitivity.
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PURPOSE: The adaptation effect in stuttering, traditionally described as the reduction of stuttering moments over repeated readings, provides a context to investigate fluency facilitation as well as a relatively controlled means of comparing fluent speech in the immediate vicinity of words that were stuttered versus fluently produced. Acoustic studies have documented decreased duration of fluent speech during adaptation but rarely address changes in disfluencies or the speech preceding or following the disfluencies. This study addresses this gap in the research by documenting frequency and duration changes in both fluent and stuttered syllables. METHOD: Fifteen people who stutter read passages aloud five times in succession. Frequency and duration of fluent syllables, pauses, stuttering-like disfluencies (SLDs) and other disfluencies (ODs) were compared across the five readings. In addition, durations for syllables before and after pauses and SLDs were compared to determine if there were anticipation or carryover effects of SLDs on surrounding syllables. RESULTS: Durations measured for more than 22 000 fluent syllables, 1531 pauses, 128 ODs and 1752 SLDs. For most of the 15 participants, significant decreases in both frequency and duration of SLDs over the five readings were observed. In addition, lengthening of fluent syllables immediately preceding the disfluent syllables was observed: this pre-SLD lengthening did not change over the five readings. CONCLUSIONS: Decreased duration of SLDs across readings supports the motor practice hypothesis, which assumes that successive reading of the same text increases the efficiency of the speech motor plans resulting in less stuttering and decreased durations of the stuttering that persists. Pre-SLD lengthening merits further study, because it informs our knowledge of the time course of stuttered events and may be associated with conscious or unconscious anticipation of upcoming SLDs that does not decrease with motor practice. WHAT THIS PAPER ADDS: What is already known on this subject The frequency of stuttering-like disfluencies (SLDs) can be reduced using a variety of fluency-enhancing strategies. For example, the adaptation effect, in which a reduction of stuttered events occurs over repeated readings of the same material, has been widely studied. Previous studies have shown that durations of fluent syllables decrease during adaptation, supporting the hypothesis that repeated practice of the motor plan leads to increased fluency. However, temporal changes in disfluent syllables and syllables preceding and following SLDs have rarely been studied, so our understanding of the effect of motor practice on stuttering reduction is incomplete. What this study adds This study has two significant findings. First, stuttered disfluencies that persisted after the initial reading of the adaptation task tended to become shorter in duration. Fluently produced syllables and those that were stuttered, both of which are speech events related to motor control of articulators, were affected in a similar manner by the motor practice associated with adaptation. Second, lengthening of fluent syllables immediately preceding stuttered syllables was observed. This pre-stuttering lengthening, however, did not decrease in duration over the five readings: the mechanism that drives this anticipatory behaviour is not affected by repeated practice. What are the clinical implications of this work? People who stutter have neural differences that lead to speech motor planning and/or execution that is less efficient than that of typical speakers. The finding that stuttering is reduced and that persisting SLDs become shorter in duration over repeated readings provides evidence that motor practice can influence the manifestation of stuttering by temporarily making those specific motor plans more efficient. This may inform treatments for stuttering. The observation that fluent syllables immediately before SLDs were lengthened, and that this lengthening was not influenced by repeated practice, extends our understanding of the time course of stuttering events and may be useful in understanding anticipation and listener reactions to stuttering.
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Gagueira , Humanos , Gagueira/diagnóstico , Fala , Medida da Produção da FalaRESUMO
The COVID-19 pandemic emphasised the global value chains (GVCs) debate by focussing on whether gains from GVC participation outweigh firms associated risks of demand and supply shocks amid rising protectionism. This paper bridges the gap between the international trade and management literature by examining the impact of COVID-19 on Commonwealth countries, an area that has received scant attention in academic literature. Using the Eora database, we simulate scenarios to examine Commonwealth countries' participation in GVCs post-COVID. We draw on the transaction cost economics (TCE) theory to develop a framework that investigates whether growing protectionism, associated with reshoring, decoupling and nearshoring, could potentially affect the constellation and participation of Commonwealth countries in GVCs post-COVID. Results show that trade protectionism is likely to impact the supply chains and lead to GVC reconfiguration, which could offer opportunities for the Commonwealth countries and firms to potentially gain following the geographical redistribution of suppliers.
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Catechol estrogens (CEs) are known to be toxic metabolites and the initiators of the oncogenesis of breast cancers via forming covalent adducts with DNAs. CEs shall also react with proteins, but their cellular protein targets remain unexplored. Here, we reported the identification of protein targets of CEs in the soluble cytosol of estrogen-sensitive breast cancer cells by multiple comparative proteomics using liquid chromatography-tandem mass spectrometry (LC-MS/MS) coupled with an improved click chemistry-based workflow. Multiple comparative proteomics composed of an experimental pair (probe versus solvent) and two control pairs (solvent versus solvent and probe versus solvent without enrichment) were studied using stable isotope dimethyl labeling. The use of 4-hydroxyethynylestradiol (4OHEE2) probe with an amide-free linker coupled with on-bead digestion and redigestion of the proteins cleaved from the beads was shown to greatly improve the recovery and identification of CE-adducted peptides. A total of 310 protein targets and 40 adduction sites were repeatedly (n ≥ 2) identified with D/H (probe/solvent) ratio >4 versus only one identified with D/H >4 from the two control pairs, suggesting that our workflow imposes only a very low background. Meanwhile, multiple comparative D/H ratios revealed that CEs may downregulate many target proteins involved in the metabolism or detoxification, suggesting a negative correlation between CE-induced adduction and expression of proteins acting on the alleviation of stress-induced cellular damages. The reported method and data will provide opportunities to study the progression of estrogen metabolism-derived diseases and biomarkers.
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Neoplasias da Mama , Estrogênios de Catecol , Neoplasias da Mama/metabolismo , Cromatografia Líquida , Química Click , Citosol , Humanos , Espectrometria de Massas em Tandem , Fluxo de TrabalhoRESUMO
The rate-determining step in free radical lipid peroxidation is the propagation of the peroxyl radical, where generally two types of reactions occur: (a) hydrogen-atom transfer (HAT) from a donor to the peroxyl radical; (b) peroxyl radical addition (PRA) to a "CâC" double bond. Peroxyl radical clocks have been used to determine the rate constants of HAT reactions (kH), but no radical clock is available to measure the rate constants of PRA reactions (kadd). In this work, we modified the analytical approach on the linoleate-based peroxyl radical clock to enable the simultaneous measurement of both kH and kadd. Compared to the original approach, this new approach involves the use of a strong reducing agent, LiAlH4, to completely reduce both HAT and PRA-derived products and the relative quantitation of total linoleate oxidation products with or without reduction. The new approach was then applied to measuring the kH and kadd values for several series of organic substrates, including para- and meta-substituted styrenes, substituted conjugated dienes, and cyclic alkenes. Furthermore, the kH and kadd values for a variety of biologically important lipids were determined for the first time, including conjugated fatty acids, sterols, coenzyme Q10, and lipophilic vitamins, such as vitamins D3 and A.
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Abundant blood proteins adducted by active electrophiles are excellent markers to predict the risk of electrophile-induced toxicity. However, detecting endogenously adducted proteins by bottom-up selective (or parallel) reaction monitoring (SRM/PRM) is challenging because of the high variability in sample preparation and detection as well as low adduction levels. Here, we reported a new approach in developing PRM methods by combining intact protein measurement with standard additions to target optimal conditions for detecting catechol estrogens (CEs)-adducted human serum albumin (HSA). Blood serum was added with multiple amounts of CEs to obtain serum standards. Intact protein measurement revealed two linear ranges of adduction levels (adducted-CE/HSA): 0.34-0.42 (R2 > 0.94) and 0.81-8.54 (R2 > 0.96) against the amount of added CEs, respectively. Six adduction sites were identified by trypsin (K20, C34, K73, K281, H338, K378) or chymotrypsin (K20, C34, K378) digestion. PRM methods targeting all adducted/nonadducted peptide pairs based on chymotrypsin or trypsin digestion were developed, and the data were compared with those obtained by intact protein measurement. Correlation plots indicated that chymotrypsin-PRM leads to poor sensitivity and largely underestimated protein adduction levels. Trypsin-PRM leads to sensitive and highly correlated (R2 > 0.91) protein adduction levels with a detection limit below the endogenous level and relative standard deviation <25%. As a proof of concept, clinical serum samples were examined by trypsin-PRM, and a slightly higher adduction level was observed for the obesity group when compared with the healthy group. This is the first report on determining adduction levels of blood proteins for long-term exposure to CEs. The standard addition approach can be generally applied to protein adductomics with resolvable mass increments by intact protein measurement to accelerate the development of bottom-up methods close to the inherent limit.
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Estrogênios de Catecol/química , Espectrometria de Massas/métodos , Peptídeos/análise , Albumina Sérica/química , Cromatografia Líquida de Alta Pressão , Quimotripsina/metabolismo , Estrogênios de Catecol/metabolismo , Humanos , Espectrometria de Massas/normas , Nanotecnologia , Peptídeos/metabolismo , Peptídeos/normas , Padrões de Referência , Albumina Sérica/metabolismo , Tripsina/metabolismoRESUMO
Marine microorganisms are an invaluable source of novel active secondary metabolites possessing various biological activities. In this study, the extraction and isolation of the marine sediment Penicillium species collected in Vietnam yielded ten secondary metabolites, including sporogen AO-1 (1), 3-indolecarbaldehyde (2), 2-[(5-methyl-1,4-dioxan-2-yl)methoxy]ethanol (3), 2-[(2R-hydroxypropanoyl)amino]benzamide (4), 4-hydroxybenzandehyde (5), chrysogine (6), 3-acetyl-4-hydroxycinnoline (7), acid 1H-indole-3-acetic (8), cyclo (Tyr-Trp) (9), and 2',3'-dihydrosorbicillin (10). Their structures were identified by the analysis of 1D and 2D NMR data. Among the isolated compounds, 2-[(5-methyl-1,4-dioxan-2-yl)methoxy]ethanol (3) showed a strong inhibitory effect against Enterococcus faecalis with a minimum inhibitory concentration value of 32 µg/mL. Both 2-[(2R-hydroxypropanoyl)amino]benzamide (4) and 4-hydroxybenzandehyde (5) selectively inhibited E. coli with minimum inhibitory concentration values of 16 and 8 µg/mL, respectively. 2',3'-Dihydrosorbicillin (10) potentially inhibited α-glucosidase activity at a concentration of 2.0 mM (66.31%).
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Antibacterianos , Organismos Aquáticos , Enterococcus faecalis/crescimento & desenvolvimento , Penicillium , Antibacterianos/química , Antibacterianos/metabolismo , Antibacterianos/farmacologia , Organismos Aquáticos/química , Organismos Aquáticos/metabolismo , Estrutura Molecular , Penicillium/química , Penicillium/metabolismo , VietnãRESUMO
Catechol estrogens (CEs) are metabolic electrophiles that actively undergo covalent interaction with cellular proteins, influencing molecular function. There is no feasible method to identify their binders in a living system. Herein, we developed a click chemistry-based approach using ethinylestradiol (EE2) as the precursor probe coupled with quantitative proteomics to identify protein targets of CEs and classify their binding strengths. Using in situ metabolic conversion and click reaction in liver microsomes, CEs-protein complex was captured by the probe, digested by trypsin, stable isotope labeled via reductive amination, and analyzed by liquid chromatography-mass spectrometry (LC-MS). A total of 334 liver proteins were repeatedly identified ( n ≥ 2); 274 identified proteins were classified as strong binders based on precursor mass mapping. The binding strength was further scaled by D/H ratio (activity probe/solvent): 259 strong binders had D/H > 5.25; 46 weak binders had 5.25 > D/H > 1; 5 nonspecific binders (keratins) had D/H < 1. These results were confirmed using spiked covalent control (strong binder) and noncovalent control (weak binder), as well as in vitro testing of cytochrome c (D/H = 5.9), which showed covalent conjugation with CEs. Many identified strong binders, such as glutathione transferase, catechol-O-methyl transferase, superoxide dismutase, catalase, glutathione peroxidase, and cytochrome c, are involved in cellular redox processes or detoxification activities. CE conjugation was shown to suppress the superoxide oxidase activity of cytochrome c, suggesting that CEs modification may alter the redox action of cellular proteins. Due to structural similarity and inert alkyne group, EE2 probe is very likely to capture protein targets of CEs in general. Thus, this strategy can be adopted to explore the biological impact of CEs modification in living systems.
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Estrogênios de Catecol/antagonistas & inibidores , Proteínas/farmacologia , Proteômica/métodos , Animais , Cromatografia Líquida , Química Click/métodos , Etinilestradiol/química , Espectrometria de Massas , Microssomos Hepáticos/química , Sondas Moleculares , Ligação Proteica , RatosRESUMO
[This corrects the article PMC10846681.].
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Lipid peroxidation is the driver of ferroptotic cell death. However, nonconjugated and conjugated polyunsaturated fatty acids potentiate ferroptosis differently, while some isoprenoid-derived lipids inhibit ferroptosis despite being highly oxidizable. In this perspective, we propose that different oxidation mechanisms and products contribute to the discrepancies in the lipids' potency in modulating ferroptosis. We first discuss the relative reactivities of various lipids toward two rate-determining free radical propagating mechanisms, hydrogen atom transfer (HAT) and peroxyl radical addition (PRA), and the resulting differential product profiles. We then discuss the role and regulation of lipid peroxidation in ferroptosis and the potential contributions of different oxidation products, such as truncated lipids and lipid electrophiles, from HAT and PRA mechanisms to the execution of ferroptosis. Lastly, we offer our perspective on the remaining questions to fully understand the process from lipid peroxidation to ferroptosis.
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Ferroptosis is a form of regulated cell death driven by lipid peroxidation of polyunsaturated fatty acids (PUFAs). Lipid peroxidation can propagate through either the hydrogen-atom transfer (HAT) or peroxyl radical addition (PRA) mechanism. However, the contribution of the PRA mechanism to the induction of ferroptosis has not been studied. In this study, we aim to elucidate the relationship between the reactivity and mechanisms of lipid peroxidation and ferroptosis induction. We found that while some peroxidation-reactive lipids, such as 7-dehydrocholesterol, vitamins D3 and A, and coenzyme Q10, suppress ferroptosis, both nonconjugated and conjugated PUFAs enhanced cell death induced by RSL3, a ferroptosis inducer. Importantly, we found that conjugated PUFAs, including conjugated linolenic acid (CLA 18:3) and conjugated linoleic acid (CLA 18:2), can induce or potentiate ferroptosis much more potently than nonconjugated PUFAs. We next sought to elucidate the mechanism underlying the different ferroptosis-inducing potency of conjugated and nonconjugated PUFAs. Lipidomics revealed that conjugated and nonconjugated PUFAs are incorporated into distinct cellular lipid species. The different peroxidation mechanisms predict the formation of higher levels of reactive electrophilic aldehydes from conjugated PUFAs than nonconjugated PUFAs, which was confirmed by aldehyde-trapping and mass spectrometry. RNA sequencing revealed that protein processing in the endoplasmic reticulum and proteasome are among the most significantly upregulated pathways in cells treated with CLA 18:3, suggesting increased ER stress and activation of unfolded protein response. These results suggest that protein damage by lipid electrophiles is a key step in ferroptosis.
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Lipid metabolism is implicated in a variety of diseases, including cancer, cell death, and inflammation, but lipidomics has proven to be challenging due to the vast structural diversity over a narrow range of mass and polarity of lipids. Isotope labeling is often used in metabolomics studies to follow the metabolism of exogenously added labeled compounds because they can be differentiated from endogenous compounds by the mass shift associated with the label. The application of isotope labeling to lipidomics has also been explored as a method to track the metabolism of lipids in various disease states. However, it can be difficult to differentiate a single isotopically labeled lipid from the rest of the lipidome due to the variety of endogenous lipids present over the same mass range. Here we report the development of a dual-isotope deuterium labeling method to track the metabolic fate of exogenous polyunsaturated fatty acids, e.g., arachidonic acid, in the context of ferroptosis using hydrophilic interaction-ion mobility-mass spectrometry (HILIC-IM-MS). Ferroptosis is a type of cell death that is dependent on lipid peroxidation. The use of two isotope labels rather than one enables the identification of labeled species by a signature doublet peak in the resulting mass spectra. A Python-based software, D-Tracer, was developed to efficiently extract metabolites with dual-isotope labels. The labeled species were then identified with LiPydomics based on their retention times, collision cross section, and m/z values. Changes in exogenous AA incorporation in the absence and presence of a ferroptosis inducer were elucidated.
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Ferroptose , Lipidômica , Lipidômica/métodos , Ácido Araquidônico , Marcação por Isótopo , Espectrometria de Massas/métodosRESUMO
INTRODUCTION: Despite the proliferation of Artificial Intelligence (AI) technology over the last decade, clinician, patient, and public perceptions of its use in healthcare raise a number of ethical, legal and social questions. We systematically review the literature on attitudes towards the use of AI in healthcare from patients, the general public and health professionals' perspectives to understand these issues from multiple perspectives. METHODOLOGY: A search for original research articles using qualitative, quantitative, and mixed methods published between 1 Jan 2001 to 24 Aug 2021 was conducted on six bibliographic databases. Data were extracted and classified into different themes representing views on: (i) knowledge and familiarity of AI, (ii) AI benefits, risks, and challenges, (iii) AI acceptability, (iv) AI development, (v) AI implementation, (vi) AI regulations, and (vii) Human - AI relationship. RESULTS: The final search identified 7,490 different records of which 105 publications were selected based on predefined inclusion/exclusion criteria. While the majority of patients, the general public and health professionals generally had a positive attitude towards the use of AI in healthcare, all groups indicated some perceived risks and challenges. Commonly perceived risks included data privacy; reduced professional autonomy; algorithmic bias; healthcare inequities; and greater burnout to acquire AI-related skills. While patients had mixed opinions on whether healthcare workers suffer from job loss due to the use of AI, health professionals strongly indicated that AI would not be able to completely replace them in their professions. Both groups shared similar doubts about AI's ability to deliver empathic care. The need for AI validation, transparency, explainability, and patient and clinical involvement in the development of AI was emphasised. To help successfully implement AI in health care, most participants envisioned that an investment in training and education campaigns was necessary, especially for health professionals. Lack of familiarity, lack of trust, and regulatory uncertainties were identified as factors hindering AI implementation. Regarding AI regulations, key themes included data access and data privacy. While the general public and patients exhibited a willingness to share anonymised data for AI development, there remained concerns about sharing data with insurance or technology companies. One key domain under this theme was the question of who should be held accountable in the case of adverse events arising from using AI. CONCLUSIONS: While overall positivity persists in attitudes and preferences toward AI use in healthcare, some prevalent problems require more attention. There is a need to go beyond addressing algorithm-related issues to look at the translation of legislation and guidelines into practice to ensure fairness, accountability, transparency, and ethics in AI.
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Algoritmos , Inteligência Artificial , Humanos , Escolaridade , Emoções , EmpatiaRESUMO
OBJECTIVE: Nonintubated uniportal video-assisted thoracoscopic surgery (VATS) is a recent controversial procedure in many countries. Hence, the authors would like to present the experience in performing this approach and evaluate its initial results in the treatment, particularly, of intrathoracic diseases in Vietnam. METHODS: A prospective, descriptive study was conducted on 17 patients with intrathoracic diseases treated with nonintubated uniportal VATS from February to July 2019. Preoperative, intraoperative, and postoperative parameters were gathered and analyzed by SPSS Statistics, Version 18.0. RESULTS: Patients had an average age of 49.2 ± 20.5 (range 6 to 71) years. Regarding operative indications and methods, there were 3 ground glass opacity (17.6%) and 1 bullous lung disease receiving wedge resection (5.9%); 8 mediastinal tumors (47.1%) having resection, and 5 non-small-cell lung cancers receiving lobectomy combined with node dissection (29.4%). The average operative time and pleural drainage time were 108.6 ± 28.17 min (range 60 to 160) and 3.7 ± 1.18 days (range 2 to 8), respectively. The average hospitalization time was 4.9 ± 1.76 (range 3 to 12) days. No mortalities or major complications were recorded postoperatively. CONCLUSIONS: Nonintubated uniportal VATS is a safe and considerable surgical choice for appropriate intrathoracic conditions.
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Carcinoma Pulmonar de Células não Pequenas , Neoplasias Pulmonares , Adolescente , Adulto , Idoso , Criança , Humanos , Pessoa de Meia-Idade , Estudos Prospectivos , Estudos Retrospectivos , Cirurgia Torácica Vídeoassistida , Vietnã/epidemiologia , Adulto JovemRESUMO
A total of 344 halophilic bacteria were isolated from fish fermentation broths, solar salt crystals, seawater, and muds from ponds of salt pans in Vietnam and subjected to aroma evaluation using fish broth containing 29 ~ 30% (w/v) NaCl. One isolate from a salt crystal with the highest aroma score was selected, identified by using 16S rDNA sequence, and named Marinococcus halotolerans SPQ. The GC-MS results of the fish broth fermented by M. halotolerans SPQ revealed elevated concentrations of several aroma compounds such as ethyl alcohol, 1-propanol, 1-butyl alcohol, 1-amyl alcohol, and methionol. During the validation tests for M. halotolerans SPQ, using 2 kg of anchovy fish in 30% (w/v) NaCl at pH 5.78, the total and amino nitrogen values in the broth increased over time from 15.2 g/L at the beginning to 26.3 g/L at 6th month, with these values being comparable to those of the control. The ammoniacal nitrogen value (2.52 g/L) in the inoculated broth at 6th month was slightly higher than that (2.21 g/L) of control. The histamine content of the fish broth inoculated with M. halotolerans SPQ after 6 months was 110.12 mg/L, less than the maximum permitted safety limit of 200 mg/L, indicating it to be safe. Physical parameters, such as the total, amino, ammoniacal nitrogens, and histamine content of fish broth fermented by M. halotolerans MPQ met the standards for Vietnamese fish sauces. Two important umami amino acids, aspartic and glutamic acid, were seen to significantly increase, by 23.5% and 35.1%, respectively, even in the extremely harsh fermentation conditions posed by 30% (w/v) NaCl. The color, odor, and taste of the fish sauce fermented by M. halotolerans SPQ elicited the highest preference score accorded by the panelists. Taken together, M. halotolerans SPQ is a promising starter culture strain for fish sauce fermentation.
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Bacillus subtilis and Escherichia coli are evolutionarily divergent model organisms whose analysis has enabled elucidation of fundamental differences between Gram-positive and Gram-negative bacteria, respectively. Despite their differences in cell cycle control at the molecular level, the two organisms follow the same phenomenological principle, known as the adder principle, for cell size homeostasis. We thus asked to what extent B. subtilis and E. coli share common physiological principles in coordinating growth and the cell cycle. We measured physiological parameters of B. subtilis under various steady-state growth conditions with and without translation inhibition at both the population and single-cell levels. These experiments revealed core physiological principles shared between B. subtilis and E. coli Specifically, both organisms maintain an invariant cell size per replication origin at initiation, under all steady-state conditions, and even during nutrient shifts at the single-cell level. Furthermore, the two organisms also inherit the same "hierarchy" of physiological parameters. On the basis of these findings, we suggest that the basic principles of coordination between growth and the cell cycle in bacteria may have been established early in evolutionary history.IMPORTANCE High-throughput, quantitative approaches have enabled the discovery of fundamental principles describing bacterial physiology. These principles provide a foundation for predicting the behavior of biological systems, a widely held aspiration. However, these approaches are often exclusively applied to the best-known model organism, E. coli In this report, we investigate to what extent quantitative principles discovered in Gram-negative E. coli are applicable to Gram-positive B. subtilis We found that these two extremely divergent bacterial species employ deeply similar strategies in order to coordinate growth, cell size, and the cell cycle. These similarities mean that the quantitative physiological principles described here can likely provide a beachhead for others who wish to understand additional, less-studied prokaryotes.
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Bacillus subtilis/fisiologia , Fenômenos Fisiológicos Bacterianos , Divisão Celular , Replicação do DNA , Origem de Replicação , Bacillus subtilis/citologia , Ciclo Celular , Escherichia coli/citologia , Escherichia coli/fisiologiaRESUMO
The title compound, C12H8O4, is a furan-ocoumarin [systematic name: 4-meth-oxy-7H-furo[3,2-g]chromen-7-one], which was isolated from the Indian herb T. stictocarpum. The mol-ecule is almost planar with an r.m.s. deviation of 0.024â Å for the hetero atoms of the fused-ring system. In the crystal, mol-ecules are linked by C-Hâ¯O hydrogen bonds, forming a three-dimensional framework. There are offset π-π inter-actions present involving the coumarin moieties stacking along the a-axis direction [shortest inter-centroid distance = 3.717â (3)â Å].
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This study aims to investigate whether overexpressing the trehalose biosynthetic gene, otsBA operon, in ß-carotene-producing recombinant Escherichia coli protects cells from toxic impurities in crude glycerol. The concentrations of potassium and methanol in crude glycerol were too low to inhibit cell growth. Cell growth and production in control cell culture were inhibited significantly in the presence of a small amount of crude fatty acids. Peroxides were generated in the presence of crude fatty acids during autoclaving and, thus, the inhibitory effect of crude fatty acids was caused primarily by these peroxides. Engineered cells overexpressing otsBA tolerated crude fatty acids (≤42 wet-g/L), methanol (≤7.5 g/L), and t-BuOOH (≤60 µM) in separate experiments and tolerated up to 60 g/L crude glycerol. These results demonstrate that overexpressing otsBA endowed cells with the capacity to tolerate the toxicity of crude glycerol for direct use.
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Proteínas de Escherichia coli/genética , Escherichia coli/fisiologia , Glucosiltransferases/genética , Glicerol/toxicidade , Monoéster Fosfórico Hidrolases/genética , Recombinação Genética , Trealose/biossíntese , beta Caroteno/biossíntese , Escherichia coli/genética , PlasmídeosRESUMO
This report describes the improved quantitative determination of valiolamine in a medium for microbial culture using high-performance liquid chromatography with UV detection. Valiolamine aqueous solution was dried, dissolved in dimethyl sulfoxide and derivatization performances of phenylisocyanate (PHI), 1-fluoro-2,4-dinitrobenznene and 1-naphthylisothiocyanate were compared in the presence of triethylamine. The PHI was chosen as the most suitable derivatization reagent and the valiolamine-PHI derivative was identified by thin-layer chromatography and electrospray ionization mass spectrometry. The derivative eluted at 10.5 min on a reverse-phase column using a mobile phase composed of 10% acetonitrile in water containing 0.5 mM sodium octyl sulfate (pH 3.0), at a column flow rate of 1.0 mL/min with UV detection at 240 nm. The optimum conditions for derivatization were a reaction temperature of 30 degrees C, reaction time of 30 min, and PHI concentration higher than 33.6 mM. Calibration curves were linear in the range of 0.99-19.95 microg/mL for the standard solutions and 24.9-99.7 microg/mL for the spiked sample. The proposed method was validated and proven to be selective, accurate and precise and suitable for the quantitative analysis of valiolamine in medium for microbial cultures.