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This study presents the vascularized tissue on mesh-assisted platform (VT-MAP), a novel microfluidic in vitro model that uses an open microfluidic principle for cultivating vascularized organoids. Addressing the gap in 3D high-throughput platforms for drug response analysis, the VT-MAP can host tumor clusters of various sizes, allowing for precise, size-dependent drug interaction assessments. Key features include capability for forming versatile co-culture conditions (EC, fibroblasts and colon cancer organoids) that enhance tumor organoid viability and a perfusable vessel network that ensures efficient drug delivery and maintenance of organoid health. The VT-MAP enables the culture and analysis of organoids across a diverse size spectrum, from tens of microns to several millimeters. The VT-MAP addresses the inconsistencies in traditional organoid testing related to organoid size, which significantly impacts drug response and viability. Its ability to handle various organoid sizes leads to results that more accurately reflect patient-derived xenograft (PDX) models and differ markedly from traditional in vitro well plate-based methods. We introduce a novel image analysis algorithm that allows for quantitative analysis of organoid size-dependent drug responses, marking a significant step forward in replicating PDX models. The PDX sample from a positive responder exhibited a significant reduction in cell viability across all organoid sizes when exposed to chemotherapeutic agents (5-FU, oxaliplatin, and irinotecan), as expected for cytotoxic drugs. In sharp contrast, PDX samples of a negative responder showed little to no change in viability in smaller clusters and only a slight reduction in larger clusters. This differential response, accurately replicated in the VT-MAP, underscores its ability to generate data that align with PDX models and in vivo findings. Its capacity to handle various organoid sizes leads to results that more accurately reflect PDX models and differ markedly from traditional in vitro methods. The platform's distinct advantage lies in demonstrating how organoid size can critically influence drug response, revealing insights into cancer biology previously unattainable with conventional techniques.
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Antineoplásicos , Neoplasias , Animais , Humanos , Telas Cirúrgicas , Antineoplásicos/farmacologia , Neoplasias/tratamento farmacológico , Neoplasias/patologia , Modelos Animais de Doenças , Organoides/patologiaRESUMO
This study addresses the demand for research models that can support patient-treatment decisions and clarify the complexities of a tumor microenvironment by developing an advanced non-animal preclinical cancer model. Based on patient-derived tumor spheroids (PDTS), the proposed model reconstructs the tumor microenvironment with emphasis on tumor spheroid-driven angiogenesis. The resulting microfluidic chip system mirrors angiogenic responses elicited by PDTS, recapitulating patient-specific tumor conditions and providing robust, easily quantifiable outcomes. Vascularized PDTS exhibited marked angiogenesis and tumor proliferation on the microfluidic chip. Furthermore, a drug that targets the vascular endothelial growth factor receptor 2 (VEGFR2, ramucirumab) was deployed, which effectively inhibited angiogenesis and impeded tumor invasion. This innovative preclinical model was used for investigating distinct responses for various drug combinations, encompassing HER2 inhibitors and angiogenesis inhibitors, within the context of PDTS. This integrated platform could potentially advance precision medicine by harmonizing diverse data points within the tumor microenvironment with a focus on the interplay between cancer and the vascular system.
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Neoplasias , Fator A de Crescimento do Endotélio Vascular , Humanos , Fator A de Crescimento do Endotélio Vascular/metabolismo , Angiogênese , Neovascularização Patológica/metabolismo , Inibidores da Angiogênese/farmacologia , Inibidores da Angiogênese/uso terapêutico , Linhagem Celular Tumoral , Neoplasias/tratamento farmacológicoRESUMO
Several functions of autophagy associated with proliferation, differentiation, and migration of endothelial cells have been reported. Due to lack of models recapitulating angiogenic sprouting, functional heterogeneity of autophagy in endothelial cells along angiogenic sprouts remains elusive. Here, we apply an angiogenesis-on-a-chip to reconstruct 3D sprouts with clear endpoints. We perform single-cell RNA sequencing of sprouting endothelial cells from our chip to reveal high activation of autophagy in two endothelial cell populations- proliferating endothelial cells in sprout basements and stalk-like endothelial cells near sprout endpoints- and further the reciprocal expression pattern of autophagy-related genes between stalk- and tip-like endothelial cells near sprout endpoints, implying an association of autophagy with tip-stalk cell specification. Our results suggest a model describing spatially differential roles of autophagy: quality control of proliferating endothelial cells in sprout basements for sprout elongation and tip-stalk cell specification near sprout endpoints, which may change strategies for developing autophagy-based anti-angiogenic therapeutics.
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Células Endoteliais , Neovascularização Fisiológica , Neovascularização Fisiológica/genética , Angiogênese , Dispositivos Lab-On-A-Chip , Análise de Sequência de RNARESUMO
Tumors disrupt the normal homeostasis of human body as they proliferate in abnormal speed. For constant proliferation, tumors recruit new blood vessels transporting nutrients and oxygen. Immune system simultaneously recruits lymphatic vessels to induce the death of tumor cells. Hence, understanding tumor dynamics are important to developing anti-cancer therapies. Tumor-on-a-chip technology can be applied to identify the structural and functional units of tumors and tumor microenvironments with high reproducibility and reliability, monitoring the development and pathophysiology of tumors, and predicting drug effectiveness. Herein, we explore the ability of tumor-on-a-chip technology to mimic angiogenic and lymphangiogenic tumor microenvironments of organs. Microfluidic systems allow elaborate manipulation of the development and status of cancer. Therefore, they can be used to validate the effects of various drug combinations, specify them, and assess the factors that influence cancer treatment. We discuss the mechanisms of action of several drugs for cancer treatment in terms of tumor growth and progression involving angiogenesis and lymphangiogenesis. Moreover, we present future applications of emerging tumor-on-a-chip technology for drug development and cancer therapy.
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Dispositivos Lab-On-A-Chip , Neoplasias , Humanos , Microfluídica , Neoplasias/tratamento farmacológico , Neoplasias/patologia , Reprodutibilidade dos Testes , Microambiente TumoralRESUMO
PURPOSE: Postoperative delirium (POD) is a common but serious complication after cardiac surgery and is associated with various short- and long-term outcomes. In this study, we investigated the effects of intraoperative glycemic variability (GV) and other glycemic variables on POD after cardiac surgery. PATIENTS AND METHODS: A retrospective single-center cohort analysis was conducted using data from electronic medical record from 2018 to 2020. A total of 705 patients undergoing coronary artery bypass graft surgery and/or valve surgery, and/or aortic replacement surgery were included in the analysis. Intraoperative GV was assessed with a coefficient of variation (CV), which was defined as the standard deviation of five intraoperative blood glucose measurements divided by the mean. POD assessment was performed three times a day in the ICU and twice a day in the ward until discharge by trained medical staff. POD was diagnosed if any of the Confusion Assessment Method for the Intensive Care Unit was positive in the ICU, and the Confusion Assessment Method was positive in the ward. Multivariable logistic regression was used to identify associations between intraoperative GV and POD. RESULTS: POD occurred in 306 (43.4%) patients. When intraoperative glycemic CV was compared as a continuous variable, the delirium group had higher intraoperative glycemic CV than the non-delirium group (22.59 [17.09, 29.68] vs 18.19 [13.00, 23.35], p < 0.001), and when intraoperative glycemic CV was classified as quartiles, the incidence of POD increased as intraoperative glycemic CV quartiles increased (first quartile 29.89%; second quartile 36.67%; third quartile 44.63%; and fourth quartile 62.64%, p < 0.001). In the multivariable logistic regression model, patients in the third quartile of intraoperative glycemic CV were 1.833 times (OR 1.833, 95% CI: 1.132-2.967, p = 0.014), and patients in the fourth quartile of intraoperative glycemic CV were 3.645 times (OR 3.645, 95% CI: 2.235-5.944, p < 0.001) more likely to develop POD than those in the first quartile of intraoperative glycemic CV. CONCLUSION: Intraoperative blood glucose fluctuation, manifested by intraoperative GV, is associated with POD after cardiac surgery. Patients with a higher intraoperative GV have an increased risk of POD.
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Procedimentos Cirúrgicos Cardíacos , Delírio , Glicemia , Procedimentos Cirúrgicos Cardíacos/efeitos adversos , Estudos de Coortes , Delírio/epidemiologia , Delírio/etiologia , Glucose , Humanos , Complicações Pós-Operatórias/epidemiologia , Estudos Retrospectivos , Fatores de RiscoRESUMO
Single-atom catalysts (SACs) hold the promise of utilizing 100% of the participating atoms in a reaction as active catalytic sites, achieving a remarkable boost in catalytic efficiency. Thus, they present great potential for noble metal-based electrochemical application systems, such as water electrolyzers and fuel cells. However, their practical applications are severely hindered by intrinsic complications, namely atom agglomeration and relocation, originating from the uncontrollably high surface energy of isolated single-atoms (SAs) during postsynthetic treatment processes or catalytic reactions. Extensive efforts have been made to develop new methodologies for strengthening the interactions between SAs and supports, which could ensure the desired stability of the active catalytic sites and their full utilization by SACs. This review covers the recent progress in SACs development while emphasizing the association between the regulation of coordination environments (e.g., coordination atoms, numbers, sites, structures) and the electrocatalytic performance of the hydrogen evolution reaction (HER) and oxygen evolution reaction (OER). The crucial role of coordination chemistry in modifying the intrinsic properties of SACs and manipulating their metal-loading, stability, and catalytic properties is elucidated. Finally, the future challenges of SACS development and the industrial outlook of this field are discussed.
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Hidrogênio , Oxigênio , Catálise , Metais/química , ÁguaRESUMO
BACKGROUND: Somatic tissue oxygen saturation (SstO2) is associated with systemic hypoperfusion. Kidney dysfunction may lead to increased mortality and morbidity in patients who undergo living donor liver transplantation (LDLT). We investigated the clinical utility of SstO2 during LDLT for identifying postoperative kidney dysfunction. PATIENTS AND METHODS: Data from 304 adults undergoing elective LDLT between January 2015 and February 2020 at Seoul St. Mary's Hospital were retrospectively collected. Thirty-six patients were excluded based on the exclusion criteria. In total, 268 adults were analyzed, and 200 patients were 1:1 propensity score (PS)-matched. RESULTS: Patients with early kidney dysfunction had significantly lower intraoperative SstO2 values than those with normal kidney function. Low SstO2 (< 66%) 1 h after graft reperfusion was more highly predictive of early kidney dysfunction than the values measured in other intraoperative phases. A decline in the SstO2 was also related to kidney dysfunction. CONCLUSIONS: Kidney dysfunction after LDLT is associated with patient morbidity and mortality. Our results may assist in the detection of early kidney dysfunction by providing a basis for analyzing SstO2 in patients undergoing LDLT. A low SstO2 (< 66%), particularly 1 h after graft reperfusion, was significantly associated with early kidney dysfunction after surgery. SstO2 monitoring may facilitate the identification of early kidney dysfunction and enable early management of patients.
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Nefropatias , Rim/metabolismo , Transplante de Fígado , Doadores Vivos , Saturação de Oxigênio , Complicações Pós-Operatórias , Feminino , Humanos , Nefropatias/sangue , Nefropatias/etiologia , Nefropatias/mortalidade , Masculino , Pessoa de Meia-Idade , Complicações Pós-Operatórias/sangue , Complicações Pós-Operatórias/etiologia , Complicações Pós-Operatórias/mortalidade , Estudos Retrospectivos , Seul/epidemiologiaRESUMO
BACKGROUND: Robot-assisted laparoscopic prostatectomy (RALP) is a favored surgical approach for treating prostate cancer. However, RALP does not decrease postoperative pain significantly despite its minimal invasiveness. The pain associated with robot-assisted surgery is most severe during the immediate postoperative period. We aimed to demonstrate that preoperative rectus sheath block (RSB) can reduce acute pain after RALP. METHODS: A prospective non-randomized study with two parallel groups was performed from June 2020 to August 2020. A total of 100 patients undergoing RALP were divided into two groups: the RSB group (n = 50) and the non-RSB group (n = 50). Ultrasound-guided RSB was performed preoperatively only in the RSB group. The primary outcome of the study was the visual analog scale (VAS) pain score during coughing (VAS-C) 1 h after surgery. In addition, the VAS pain score at rest (VAS-R) and the VAS-C were assessed up to 24 h after surgery. The doses of postoperative opioids consumed were also recorded. RESULTS: The RSB group had a significantly lower VAS-C 1 h after RALP (58 [47-73] vs. 74 [63-83] mm, p = 0.001). In addition, the RSB group had significantly lower VAS-R and VAS-C scores, and postoperative opioid requirement, up to 6 h after surgery compared to the non-RSB group. Moreover, the VAS-R was significantly lower in the RSB group than in the non-RSB group 24 h after surgery. CONCLUSION: Preoperative RSB significantly improved analgesia during the early period after RALP. The long-term analgesic efficacy of RSB needs further study.
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Bloqueio Nervoso , Robótica , Analgésicos Opioides/uso terapêutico , Humanos , Masculino , Manejo da Dor , Dor Pós-Operatória/tratamento farmacológico , Dor Pós-Operatória/etiologia , Dor Pós-Operatória/prevenção & controle , Estudos Prospectivos , Prostatectomia , Reto do Abdome , Ultrassonografia de IntervençãoRESUMO
Force-selective mechanochemical reactions may be important for applications in polymer mechanochemistry, yet it is difficult to achieve such reactions. This paper reports that cis-N-phthalimidoaziridine incorporated into a macromolecular backbone undergoes migration of N-phthalimido group to afford imine under mechanochemical condition and not thermal one. The imine is further hydrolyzed by water bifurcating into amine and aldehyde. These structural transformations are confirmed by 1 Hâ NMR and FT-IR spectroscopic analyses. Computational simulations are conducted for the aziridine mechanophore to propose the mechanism of reaction and define the substrate scope of reaction.
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AIM: We evaluated the prognostic value of serum- and cerebrospinal fluid (CSF)-ubiquitin carboxyl-terminal esterase L1 protein (UCHL1) measurements in post- post-out of hospital cardiac arrest (OHCA) patients treated with target temperature management (TTM), to predict neurologic outcome. METHODS: This was a prospective single-centre observational cohort study, conducted from April 2018 to September 2019. Serum- and CSF-UCHL1 were obtained immediately (UCHL1initial), 24â¯h (UCHL124), 48â¯h (UCHL148), and 72â¯h (UCHL172) after return of spontaneous circulation (ROSC). The area under the receiver operating characteristic curves (AUROC) and Delong method were used to identify cut-off values of serum- and CSF-UCHL1initial, UCHL124, UCHL148, UCHL172 for predicting neurologic outcomes. RESULTS: Of 38 patients enrolled, 16 comprised the poor outcome group. The AUROCs for serum- and CSF-UCHL1initial were 0.71 and 0.93 in predicting poor neurological outcomes, respectively (pâ¯=â¯0.01). The AUROCs for serum- and CSF-UCHL124 were 0.85 and 0.91 (pâ¯=â¯0.24). The AUROCs for serum- and CSF-UCHL148 were 0.90 and 0.97 (pâ¯=â¯0.07). The AUROCs for serum- and CSF-UCHL172 were 0.94 and 0.98 (pâ¯=â¯0.25). CONCLUSION: Findings of this study demonstrate that CSF-UCHL1 measured immediately, 24, 48, and 72â¯h after ROSC is a valuable predictor for evaluating neurologic outcomes, whereas serum-UCHL1 measured at 24, 48, and 72â¯h after ROSC showed a significant performance in the prognostication of poor outcomes in post-OHCA patients treated with TTM.
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Hipotermia Induzida , Parada Cardíaca Extra-Hospitalar , Humanos , Parada Cardíaca Extra-Hospitalar/terapia , Prognóstico , Estudos Prospectivos , Ubiquitina TiolesteraseRESUMO
The application of aziridines as nonvulnerable mechanophores is reported. Upon exposure to a mechanical force, stereochemically pure nonactivated aziridines incorporated into the backbone of a macromolecule do not undergo cis-trans isomerization, thus suggesting retention of the ring structure under force. Nonetheless, aziridines react with a dipolarophile and seem not to obey conventional reaction pathways that involve C-C or C-N bond cleavage prior to the cycloaddition. Our work demonstrates that a nonvulnerable chemical structure can be a mechanophore.
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2'-Fucosyllactose (2-FL), one of the most abundant oligosaccharides in human milk, has been spotlighted for its neutraceutical and pharmaceutical potentials. Microbial production of 2-FL is promising since it is efficient as compared to other production methods. In 2-FL microbial production via the salvage pathway for biosynthesis of guanosine 5'-diphosphate (GDP)-l-fucose from fucose, the conversion yield from fucose is important because of the high price of fucose. In this study, deletion of the genes (araA and rhaA) coding for arabinose isomerase (AraA) and rhamnose isomerase (RhaA) was attempted in engineered Escherichia coli for improving 2-FL production by using fucose, lactose, and glycerol. The engineered E. coli constructed previously is able to express fucokinase/GDP-l-fucose pyrophosphorylase (Fkp) from Bacteroides fragilis and the α-1,2-fucosyltransferase (FucT2) from Helicobacter pylori and deficient in ß-galactosidase (LacZ), fucose isomerase (FucI), and fuculose kinase (FucK). The additional double-deletion of the araA and rhaA genes in the engineered E. coli enhanced the product yield of 2-FL to 0.52 mole 2-FL/mole fucose, and hence the concentration of 2-FL reached to 47.0 g/L, which are 44% and two-fold higher than those (23.1 g/L and 0.36 mole 2-FL/mole fucose) of the control strain in fed-batch fermentation. Elimination of sugar isomerases exhibiting promiscuous activities with fucose might be critical in the microbial production of 2-FL through the salvage pathway of GDP-l-fucose.
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Proteínas de Escherichia coli , Escherichia coli , Fucose/metabolismo , Deleção de Genes , Engenharia Metabólica , Trissacarídeos/biossíntese , Escherichia coli/genética , Escherichia coli/metabolismo , Proteínas de Escherichia coli/genética , Proteínas de Escherichia coli/metabolismo , Fucose/genética , Trissacarídeos/genéticaRESUMO
3-Fucosyllactose (3-FL), one of the major oligosaccharides in human breast milk, is produced in engineered Escherichia coli. In order to search for a good α-1,3-fucosyltransferase, three bacterial α-1,3-fucosyltransferases are expressed in engineered E. coli deficient in ß-galactosidase activity and expressing the essential enzymes for the production of guanosine 5'-diphosphate-l-fucose, the donor of fucose for 3-FL biosynthesis. Among the three enzymes tested, the fucT gene from Helicobacter pylori National Collection of Type Cultures 11637 gives the best 3-FL production in a simple batch fermentation process using glycerol as a carbon source and lactose as an acceptor. In order to use glucose as a carbon source, the chromosomal ptsG gene, considered the main regulator of the glucose repression mechanism, is disrupted. The resulting E. coli strain of ∆LP-YA+FT shows a much lower performance of 3-FL production (4.50 g L-1 ) than the ∆L-YA+FT strain grown in a glycerol medium (10.7 g L-1 ), suggesting that glycerol is a better carbon source than glucose. Finally, the engineered E. coli ∆LW-YA+FT expressing the essential genes for 3-FL production and blocking the colanic acid biosynthetic pathway (∆wcaJ) exhibits the highest concentration (11.5 g L-1 ), yield (0.39 mol mol-1 ), and productivity (0.22 g L-1 h) of 3-FL in glycerol-limited fed-batch fermentation.
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Escherichia coli/metabolismo , Fucosiltransferases/metabolismo , Helicobacter pylori/enzimologia , Oligossacarídeos/metabolismo , Escherichia coli/genética , Fucose/metabolismo , Fucosiltransferases/genética , Lactose/metabolismoRESUMO
L-Fucose (6-deoxy-L-galactose) is a major constituent of glycans and glycolipids in mammals. Fucosylation of glycans can confer unique functional properties and may be an economical way to manufacture L-fucose. Research can extract L-fucose directly from brown algae, or by enzymatic hydrolysis of L-fucose-rich microbial exopolysaccharides. However, these L-fucose production methods are not economical or scalable for various applications. We engineered an Escherichia coli strain to produce L-fucose. Specifically, we modified the strain genome to eliminate endogenous L-fucose and lactose metabolism, produce 2'-fucosyllactose (2'-FL), and to liberate L-fucose from 2'-FL. This E. coli strain produced 16.7 g/L of L-fucose with productivity of 0.1 g·L-1 ·h-1 in a fed-batch fermentation. This study presents an efficient one-pot biosynthesis strategy to produce a monomeric form of L-fucose by microbial fermentation, making large-scale industrial production of L-fucose feasible.
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Técnicas de Cultura Celular por Lotes/métodos , Escherichia coli/metabolismo , Fucose/metabolismo , Reatores Biológicos , Vias Biossintéticas , Escherichia coli/genética , Fermentação , Fucose/genética , Microbiologia Industrial/métodos , Engenharia Metabólica/métodosRESUMO
Fucosyllactoses (FLs), present in human breast milk, have been reported to benefit human health immensely. Especially, 3-fucosyllactose (3-FL) has numerous benefits associated with a healthy gut ecosystem. Metabolic engineering of microorganisms is thought to be currently the only option to provide an economically feasible route for large-scale production of 3-FL. However, engineering principles for α-1,3-fucosyltransferases (1,3-FTs) are not well-known, resulting in the lower productivity of 3-FL than that of 2'-fucosyllactose (2'-FL), although both 2'-FL and 3-FL follow a common pathway to produce GDP-L-fucose. The C-terminus of 1,3-FTs is composed of heptad repeats, responsible for dimerization of the enzymes, and a peripheral membrane anchoring region. It has long been thought that truncation of most heptad repeats, retaining just 1 or 2, helps the soluble expression of 1,3-FTs. However, whether the introduction of truncated version of 1,3-FTs enhances the production of 3-FL in a metabolically engineered strain, is yet to be tested. In this study, the effect of these structural components on the production of 3-FL in Escherichia coli was evaluated through systematic truncation and elongation of the C-terminal regions of three 1,3-FTs from Helicobacter pylori. Although these three 1,3-FTs contained heptad repeats and membrane-anchoring regions of varying lengths, they commonly exhibited an optimal performance when the number of heptad repeats was increased, and membrane-binding region was removed. The production of 3-FL could be increased 10-20-fold through this simple strategy.
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Proteínas de Bactérias , Escherichia coli , Fucosiltransferases , Helicobacter pylori/genética , Lactose , Engenharia Metabólica , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Escherichia coli/enzimologia , Escherichia coli/genética , Fucosiltransferases/biossíntese , Fucosiltransferases/genética , Helicobacter pylori/enzimologia , Humanos , Lactose/análogos & derivados , Lactose/biossíntese , Lactose/genética , Engenharia de ProteínasRESUMO
2'-Fucosyllactose (2-FL) is one of most abundant oligosaccharides in human milk, which is involved in many biological functions for infant health. Since 2-FL has a great potential in application to functional food materials and pharmaceuticals, several microbial systems for mass production of 2-FL have been developed in recent years. Microbial production of 2-FL was suggested to be influenced by a number of factors including fucosylation activity of α-1,2-fucosyltransferase. In the present study, the wcfB gene coding for α-1,2-fucosyltransferase from Bacteroides fragilis was screened from eleven candidates of putative α-1,2-fucosyltransferase. Introduction of the wcfB gene allows the lacZ-deleted strain of E. coli expressing the genes for guanosine 5'-diphosphate (GDP)-l-fucose biosynthetic enzymes to produce 2-FL. As a result of fed-batch fermentation, 15.4g/L extracellular concentration of 2-FL with 2-FL yield of 0.858g/g lactose and productivity of 0.530g/L/h were obtained. In addition, the feasibility of industrial production of 2-FL using this microbial system was demonstrated by performing fed-batch fermentation in a 75L bioreactor.
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Bacteroides fragilis/enzimologia , Bacteroides fragilis/genética , Escherichia coli/genética , Fucosiltransferases/genética , Fucosiltransferases/metabolismo , Trissacarídeos/biossíntese , Proteínas de Bactérias/genética , Técnicas de Cultura Celular por Lotes , Reatores Biológicos , Fermentação , Fucose , Regulação Bacteriana da Expressão Gênica , Engenharia Genética , Vetores Genéticos , Óperon Lac , Leite Humano/química , Recombinação Genética , Deleção de SequênciaRESUMO
Orthogonal frequency division multiple access-based passive optical network (OFDMA-PON) is considered as a strong candidate for next-generation optical access network. In intensity modulation/direct detection system, OFDMA-PON downlink transmission is relatively stable, but critical issues exist in uplink multiple access. Because of different optical paths, optical beat interference (OBI) and timing offset effect are generated, which seriously disturb signal detection. We propose optical pulse division multiplexing-based OBI reduction. By considering both the spectrum broadening effect and the time domain near orthogonality, OBI could be reduced. We demonstrate that the spectral efficiency can be improved from 0.37 to 3.8 bit/s/Hz in 1-GHz signal bandwidth.
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Whole-cell biotransformation of fatty acids can be influenced by the activities of catalytic enzymes and by the efficiency of substrate transport into host cells. Here, we improved fatty acid hydration activity of the recombinant Escherichia coli expressing an oleate hydratase of Stenotrophomonas maltophilia by targeting the catalytic enzyme into the periplasm instead of the cytoplasm. Recombinant E. coli producing OhyA in the periplasm under guidance of the PelB signal sequence (E. coli OhyA_PP) exhibited significantly greater hydration activity with oleic acid and linoleic acid compared to a recombinant E. coli producing OhyA in the cytoplasm (E. coli OhyA_CS). For example, the oleate double bond hydration rate of E. coli OhyA_PP was >400 µmol/g dry cells/min (400 U/g dry cells), which is >10-fold higher than that of E. coli OhyA_CS. As the specific activities of the enzymes targeted into the cytoplasm and periplasm were comparable, we assumed that targeting OhyA into the periplasm could accelerate fatty acid transport to the catalytic enzymes by skipping the major mass transport barrier of the cytoplasmic membrane. Our results will contribute to the development of whole-cell biocatalysts for fatty acid biotransformation.
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Enzimas/metabolismo , Escherichia coli/genética , Ácidos Graxos/metabolismo , Periplasma/enzimologia , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Bioengenharia , Citoplasma/metabolismo , Enzimas/genética , Escherichia coli/enzimologia , Ácido Oleico/metabolismo , Polissacarídeo-Liases/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismoRESUMO
In orthogonal frequency division multiple access based passive optical network (OFDMA-PON) uplink, synchronization between optical network units (ONUs) is very important to maintain orthogonality. The synchronization among uplink signals is considered as one of the main challenges in OFDMA-PON due to optical path difference. In this paper, the performance degradation according to timing offset between ONUs is experimentally analyzed. And we propose and demonstrate timing offset effect reduction in asynchronous multiple access by using CP extension and filter bank based multicarrier (FBMC) system in intensity modulation/direct detection (IM/DD) based OFDMA-PON uplink transmission.
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A novel technique for mitigating the optical beat interference (OBI) noise in an optical orthogonal frequency division multiple access passive optical network (OFDMA-PON) uplink transmission is presented. By using an out of signal band RF clipping tone to the optical seed carrier, the OBI noise has been reduced and the resulting throughput and spectral efficiency has been improved. As an experimental verification, we demonstrate that the spectral efficiency of 23 km and 50 km have been doubled in the OFDMA-PON uplink transmission.