De Novo Biosynthesis of Difucosyllactose by Artificial Pathway Construction and α1,3/4-Fucosyltransferase Rational Design in Escherichia coli.

Difucosyllactose (DFL) is a significant and plentiful oligosaccharide found in human breast milk. In this study, an artificial metabolic pathway of DFL was designed, focusing on the de novo biosynthesis of GDP-fucose from only glycerol. This was achieved by engineering Escherichia coli to endogenously overexpress genes manB, manC, gmd, and wcaG and heterologously overexpress a pair of fucosyltransferases to produce DFL from lactose. The introduction of α-1,2-fucosyltransferase from Helicobacter pylori (FucT2) along with α-1,3/4-fucosyltransferase (HP3/4FT) addressed rate-limiting challenges in enzymatic catalysis and allowed for highly efficient conversion of lactose into DFL. Based on these results, molecular modification of HP3/4FT was performed based on computer-assisted screening and structure-based rational design. The best-performing mutant, MH5, containing a combination of five mutated sites (F49K/Y131D/Y197N/E338D/R369A) of HP3/4FT was obtained. The best strain BLC09-58 harboring MH5 yielded 45.81 g/L of extracellular DFL in 5-L fed-batch cultures, which was the highest titer reported to date.

Engineering Escherichia coli MG1655 for Highly Efficient Biosynthesis of 2'-Fucosyllactose by De Novo GDP-Fucose Pathway.

2'-Fucosyllactose (2'-FL), the most typical human milk oligosaccharide, is used as an additive in premium infant formula. Herein, we constructed two highly effective 2'-FL synthesis producers via a de novo GDP-fucose pathway from engineered Escherichia coli MG1655. First, lacZ and wcaJ, two competitive pathway genes, were disrupted to block the invalid consumption of lactose and GDP-fucose, respectively. Next, the lacY gene was strengthened by switching its native promoter to PJ23119. To enhance the supply of endogenous GDP-fucose, the promoters of gene clusters manC-manB and gmd-fcl were strengthened individually or in combination. Subsequently, chromosomal integration of a constitutive PJ23119 promoter-based BKHT expression cassette (PJ23119-BKHT) was performed in the arsB and recA loci. The most productive plasmid-based and plasmid-free strains produced 76.9 and 50.1 g/L 2'-FL by fed-batch cultivation, respectively. Neither of them generated difucosyl lactose nor 3-fucosyllactose as byproducts.

Electrical programmable multilevel nonvolatile photonic random-access memory.

Photonic Random-Access Memories (P-RAM) are an essential component for the on-chip non-von Neumann photonic computing by eliminating optoelectronic conversion losses in data links. Emerging Phase-Change Materials (PCMs) have been showed multilevel memory capability, but demonstrations still yield relatively high optical loss and require cumbersome WRITE-ERASE approaches increasing power consumption and system package challenges. Here we demonstrate a multistate electrically programmed low-loss nonvolatile photonic memory based on a broadband transparent phase-change material (Ge2Sb2Se5, GSSe) with ultralow absorption in the amorphous state. A zero-static-power and electrically programmed multi-bit P-RAM is demonstrated on a silicon-on-insulator platform, featuring efficient amplitude modulation up to 0.2 dB/µm and an ultralow insertion loss of total 0.12 dB for a 4-bit memory showing a 100× improved signal to loss ratio compared to other phase-change-materials based photonic memories. We further optimize the positioning of dual microheaters validating performance tradeoffs. Experimentally we demonstrate a half-a-million cyclability test showcasing the robust approach of this material and device. Low-loss photonic retention-of-state adds a key feature for photonic functional and programmable circuits impacting many applications including neural networks, LiDAR, and sensors for example.

Recent progress in fucosylated derivatives of lacto-N-tetraose and lacto-N-neotetraose.

Human milk oligosaccharides (HMOs) have attracted considerable attention owing to their unique physiological functions. Two important tetrasaccharides, lacto-N-tetraose (LNT) and lacto-N-neotetraose (LNnT), are core structures of HMOs. Their safety has been evaluated and they can be added to infant formula as functional ingredients. The fucosylated derivatives of LNT and LNnT, mainly lacto-N-fucopentaose (LNFP) I, LNFP II, LNFP III, and lacto-N-difucohexaose I, exhibit prominent physiological characteristics, including modificating the intestinal microbiota, immunomodulation, anti-bacterial activities, and antiviral infection. However, they have received lesser attention than 2'-fucosyllactose. As precursors, LNT and LNnT are connected to one or two fucosyl units through α1,2/3/4 glycosidic bonds, forming a series of compounds with complex structures. These complex fucosylated oligosaccharides can be biologically synthesized using enzymatic and cell factory approaches. This review summarizes the occurrence, physiological effects, and biosynthesis of fucosylated LNT and LNnT derivatives and their future development.

Indole-3-Propionic Acid, a Gut Microbiota Metabolite, Protects Against the Development of Postoperative Delirium.

OBJECTIVE: The aim was to determine preoperative gut microbiota metabolites that may be associated with postoperative delirium (POD) development in patients and further study in rodents. SUMMARY BACKGROUND DATA: POD occurs in 9% to 50% of older patients undergoing anesthesia/surgery but lacks effective treatments or prevention. High-throughput metabolomics using liquid chromatography with tandem mass spectrometry has accelerated disease-related biomarkers discovery. We performed metabolomic studies in humans to identify potential metabolite biomarkers linked to POD and examined potential mechanisms in rodents. METHODS: We performed a prospective observational cohort study to examine the metabolomic changes that were associated with the development of POD. Then the gut microbiota-related metabolomic changes were recapitulated by gut microbiota perturbation in rodents. POD was assessed in mice using a battery of behavioral tests including novel objective test, Y-maze test, open-field test, and buried food test. The mechanisms through which gut microbiota-related metabolomic changes influenced POD were examined using chemogenetics. RESULTS: Indole-3-propionic acid (IPA) is a gut microbiota metabolite that belongs to the indole family. Baseline plasma levels of IPA were significantly inversely correlated with the onset of POD in 103 (17 cases) human individuals. This relationship was validated in preclinical mouse models for POD: reducing IPA levels through gut microbiota perturbation promoted POD-like behavior. More importantly, IPA administration deterred POD-like behavior. Colonization of germ-free mice with mutant Clostridium sporogenes that did not produce IPA-promoted POD-like behavior. Chemogenetic studies revealed that the protective effect of IPA in mice was mediated, in part, by peroxisome proliferator-activated receptor gamma coactivator 1-alpha in hippocampal interneurons. CONCLUSIONS: Gut microbiota-derived IPA is an important molecule implicated in the pathogenesis of POD, which could potentially be harnessed for POD prevention.

Phase-Transition Microcavity Laser.

Liquid-crystal microcavity lasers have attracted considerable attention because of their extraordinary tunability and sensitive response to external stimuli, and because they operate generally within a specific phase. Here, we demonstrate a liquid-crystal microcavity laser operated in the phase transition in which the reorientation of liquid-crystal molecules occurs from aligned to disordered states. A significant wavelength shift of the microlaser is observed, resulting from the dramatic changes in the refractive index of liquid-crystal microdroplets during the phase transition. This phase-transition microcavity laser is then exploited for sensitive thermal sensing, enabling a two-order-of-magnitude enhancement in sensitivity compared with the nematic-phase microlaser operated far from the transition point. Experimentally, we demonstrate an exceptional sensitivity of -40 nm/K and an ultrahigh resolution of 320 µK. The phase-transition microcavity laser features compactness, softness, and tunability, showing great potential for high-performance sensors, optical modulators, and soft matter photonics.

Assessment of Vitamin D status and association with inflammation: Biomarkers Reflecting Inflammation and Nutritional Determinants of Anemia (BRINDA) project.

BACKGROUND: It is unclear whether 25(OH)D concentrations in children and female adults may be influenced by inflammation and thus require adjustment when estimating the population prevalence of vitamin D deficiency. OBJECTIVES: We examined correlations between inflammation biomarkers, CRP or alpha-1-acid glycoprotein (AGP), and serum 25(OH)D concentrations among preschool children (PSC; 6-59 mo) and nonpregnant females of reproductive age (FRA; 15-49 y). METHODS: We analyzed cross-sectional data from 6 nationally representative nutrition surveys (Afghanistan, Cambodia, Pakistan, UK, USA, and Vietnam) conducted among PSC (n = 9880) and FRA (n = 14,749) from the Biomarkers Reflecting Inflammation and Nutritional Determinants of Anemia project. Rank correlations between CRP or AGP and 25(OH)D concentrations were examined while taking into account complex survey design effects. RESULTS: Among both PSC and FRA, correlations between inflammation and vitamin D biomarkers were weak and inconsistent across surveys. For PSC, correlation coefficients between CRP and 25(OH)D concentrations ranged from -0.04 to 0.08, and correlations between AGP and 25(OH)D ranged from 0.01 to 0.05. Correlation coefficients between CRP and 25(OH)D for FRA ranged from -0.11 to 0.14, and correlations between AGP and 25(OH)D concentrations ranged from -0.05 to 0.01. CONCLUSIONS: Based on the weak and inconsistent correlations between CRP or AGP and 25(OH)D, there is no rationale to adjust for these inflammation biomarkers when estimating population prevalence of vitamin D deficiency in PSC or FRA.

Biosynthesis of Human Milk Oligosaccharides: Enzyme Cascade and Metabolic Engineering Approaches.

Human milk oligosaccharides (HMOs) have unique beneficial effects for infants and are considered as the new gold standard for premium infant formula. They are a collection of unconjugated glycans, and more than 200 distinct structures have been identified. Generally, HMOs are enzymatically produced by elongation and/or modification from lactose via stepwise glycosylation. Each glycosylation requires a specific glycosyltransferase (GT) and the corresponding nucleotide sugar donor. In this review, the typical HMO-producing GTs and the one-pot multienzyme modules for generating various nucleotide sugar donors are introduced, the principles for designing the enzyme cascade routes for HMO synthesis are described, and the important metabolic engineering strategies for mass production of HMOs are also reviewed. In addition, the future research directions in biotechnological production of HMOs were prospected.

Microbial Synthesis of l-Fucose with High Productivity by a Metabolically Engineered Escherichia coli.

l-Fucose is a natural deoxy hexose found in a variety of organisms. It possesses many physiological effects and has potential applications in pharmaceutical, cosmetic, and food industries. Microbial synthesis via metabolic engineering attracts increasing attention for efficient production of important chemicals. Previously, we reported the construction of a metabolically engineered Escherichia coli strain with high 2'-fucosyllactose productivity. Herein, we further introduced Bifidobacterium bifidum α-l-fucosidase via both plasmid expression and genomic integration and blocked the l-fucose assimilation pathway by deleting fucI, fucK, and rhaA. The highest l-fucose titers reached 6.31 and 51.05 g/L in shake-flask and fed-batch cultivation, respectively. l-Fucose synthesis was little affected by lactose added, and there was almost no 2'-fucosyllactose residue throughout the cultivation processes. The l-fucose productivity reached 0.76 g/L/h, indicating significant potential for large-scale industrial applications.

Dissipative Acousto-optic Interactions in Optical Microcavities.

We propose and demonstrate experimentally the strong dissipative acousto-optic interaction between a suspended vibrating microfiber and a whispering-gallery microcavity. On the one hand, the dissipative response driven by an external stimulus of acoustic waves is found to be stronger than the dispersive response by 2 orders of magnitude. On the other hand, dead points emerge with the zero dissipative response at certain parameters, promising the potentials in physical sensing such as precise measurements of magnetic field and temperature. The strong dissipative acousto-optic interaction is then explored for ultrasensitive detection of broadband acoustic waves. A noise equivalent pressure as low as 0.81 Pa at 140 kHz in air is demonstrated experimentally, insensitive to cavity Q factors and does not rely on mechanical resonances.

Evolution of morphology and cohesive force of hydrate particles in the presence/absence of wax.

In the exploitation of deep-sea oil and gas resources, the multiphase production and transportation process is frequently plagued by pipeline blockage issues. Especially when hydrates and wax coexist simultaneously, the viscosity and plugging tendency of multiphase flow systems will synergistically increase. Understanding the evolution of morphology of hydrate particles and the agglomeration characteristics of hydrate particles in the presence or absence of wax crystals is crucial to flow assurance industry. With the assistance of a visualized reactor equipped with a three axis moving platform, microscopic images of cyclopentane hydrate during hydrate growth were obtained, and the cohesive force between hydrate particles was measured. It was found that during the hydrate growth on wax-free water droplets, the untransformed water inside the particles gradually wetted the surface of the particle. With the increase in temperature and contact time, the shell of hydrate particles changed from solid and rough to smooth and moist. The cohesive force measured in this work ranges from 3.14 ± 0.52 to 11.77 ± 0.68 mN m-1 with different contact times and temperature. When the contact time was 0 s and 10 s, the cohesive force between particles increased first and then stabilized with temperature. When the contact time was 20 s, the cohesive force was greater than the first two cases and showed an overall stable trend. An interesting phenomenon was also discerned: a large water bridge between particles formed during their separation process. For the wax-containing system, it required a longer time for water droplets to be converted into hydrate particles than that for wax-free systems. After wax participated in hydrate growth, hydrate particles showed the properties of elasticity and stickiness, which resulted in a larger liquid bridge between hydrate particles after their contact. It was suggested that wax crystal would alter the shell structure of hydrate particles, and change the surface properties of hydrate particles and the formation process of the liquid bridge, leading to significant and rapid increase in the cohesive force.

BIM-based task and motion planning prototype for robotic assembly of COVID-19 hospitalisation light weight structures.

Fast transmission of COVID-19 led to mass cancelling of events to contain the virus outbreak. Amid lockdown restrictions, a vast number of construction projects came to a halt. Robotic platforms can perform construction projects in an unmanned manner, thus ensuring the essential construction tasks are not suspended during the pandemic. This research developed a BIM-based prototype, including a task planning algorithm and a motion planning algorithm, to assist in the robotic assembly of COVID-19 hospitalisation light weight structures with prefabricated components. The task planning algorithm can determine the assembly sequence and coordinates for various types of prefabricated components. The motion planning algorithm can generate robots' kinematic parameters for performing the assembly of the prefabricated components. Testing of the prototype finds that it has satisfactory performance in terms of 1) the reasonableness of assembly sequence determined, 2) reachability for the assembly coordinates of prefabricated components, and 3) capability to avoid obstacles.

Design of Electronic Nose Detection System for Apple Quality Grading Based on Computational Fluid Dynamics Simulation and K-Nearest Neighbor Support Vector Machine.

Apples are one of the most widely planted fruits in the world, with an extremely high annual production. Several issues should be addressed to avoid the damaging of samples during the quality grading process of apples (e.g., the long detection period and the inability to detect the internal quality of apples). In this study, an electronic nose (e-nose) detection system for apple quality grading based on the K-nearest neighbor support vector machine (KNN-SVM) was designed, and the nasal cavity structure of the e-nose was optimized by computational fluid dynamics (CFD) simulation. A KNN-SVM classifier was also proposed to overcome the shortcomings of the traditional SVMs. The performance of the developed device was experimentally verified in the following steps. The apples were divided into three groups according to their external and internal quality. The e-nose data were pre-processed before features extraction, and then Principal Component Analysis (PCA) and Linear Discriminant Analysis (LDA) were used to reduce the dimension of the datasets. The recognition accuracy of the PCA-KNN-SVM classifier was 96.45%, and the LDA-KNN-SVM classifier achieved 97.78%. Compared with other commonly used classifiers, (traditional KNN, SVM, Decision Tree, and Random Forest), KNN-SVM is more efficient in terms of training time and accuracy of classification. Generally, the apple grading system can be used to evaluate the quality of apples during storage.

Physiological effects, biosynthesis, and derivatization of key human milk tetrasaccharides, lacto-N-tetraose, and lacto-N-neotetraose.

Human milk oligosaccharides (HMOs) have recently attracted ever-increasing interest because of their versatile physiological functions. In HMOs, two tetrasaccharides, lacto-N-tetraose (LNT) and lacto-N-neotetraose (LNnT), constitute the essential components, each accounting 6% (w/w) of total HMOs. Also, they serve as core structures for fucosylation and sialylation, generating functional derivatives and elongation generating longer chains of core structures. LNT, LNnT, and their fucosylated and/or sialylated derivatives account for more than 30% (w/w) of total HMOs. For derivatization, LNT and LNnT can be modified into a series of complex fucosylated and/or sialylated HMOs by transferring fucose residues at α1,2-, α1,3-, and α1,3/4-linkage and/or sialic acid residues at α2,3- and α2,6-linkage. Such structural diversity allows these HMOs to possess great commercial value and an application potential in the food and pharmaceutical industries. In this review, we first elaborate the physiological functions of these tetrasaccharides and derivatives. Next, we extensively review recent developments in the biosynthesis of LNT, LNnT, and their derivatives in vitro and in vivo by employing advanced enzymatic reaction systems and metabolic engineering strategies. Finally, future perspectives in the synthesis of these HMOs using enzymatic and metabolic engineering approaches are presented.

Total Synthesis of Pagoamide A.

The first total synthesis of the thiazole-containing cyclic depsipeptide pagoamide A, is detailed. The longest linear sequence of the liquid-phase synthesis comprises 9 long linear steps from simple known starting materials, which led to the unambiguous structural confirmation of pagoamide A.

A Novel ß-1,4-Galactosyltransferase from Histophilus somni Enables Efficient Biosynthesis of Lacto-N-Neotetraose via Both Enzymatic and Cell Factory Approaches.

Human milk oligosaccharides (HMOs) attract particular attention because of their health benefits for infants. Lacto-N-neotetraose (LNnT) is one of the most abundant neutral core structures of HMOs. Bacterial ß-1,4-galactosyltransferase (ß-1,4-GalT) displays an irreplaceable role in the practical application of LNnT biosynthesis. In this study, a novel ß-1,4-GalT from Histophilus somni was identified to efficiently synthesize LNnT from UDP-Gal and lacto-N-triose II (LNT II). The optimum pH and temperature were determined to be pH 6.0 and 30 °C, respectively. The enzyme showed both transgalactosylation and hydrolysis activity, with a specific activity of 3.7 and 6.6 U/mg, respectively. LNnT was synthesized using H. somni ß-1,4-GalT via both enzymatic and cell factory approaches, and both approaches provided an LNnT ratio with the remaining LNT II at approximately 1:2 when reactions attained a balance. These findings indicated that H. somni ß-1,4-GalT has a potential in biosynthesis of LNnT and derivatives in future.

Metabolic Engineering of Escherichia coli for Lacto-N-triose II Production with High Productivity.

Lacto-N-triose II (LNT II), a core structural unit of human milk oligosaccharides (HMOs), has attracted substantial attention for its nutraceutical potentials and applications in the production of complex HMOs. In this study, Escherichia coli was metabolically engineered to efficiently produce LNT II using glycerol as a carbon source and lactose as a substrate. The UDP-N-acetylglucosamine (UDP-GlcNAc) biosynthesis pathway was strengthened, and ß-1,3-N-acetylglucosaminyltransferase (LgtA) was introduced to construct an LNT II-producing base strain. To increase the titer and yield of LNT II, combinatorial optimization of the copy number and the ribosomal binding site sequence was performed to tune the gene expression strength and translation rates of the pathway enzymes. Next, multipoint mutations were introduced to glucosamine-6-phosphatesynthase (GlmS) to relieve the feedback inhibition. Then, a series of engineered strains were constructed by blocking the futile pathways by the deletion of the relevant genes. Finally, the culture conditions were optimized. LNT II titer was improved step-by-step from 0.53 to 5.52 g/L in shake-flask cultivations. Fed-batch culture of the final engineered strain produced 46.2 g/L of LNT II, with an LNT II productivity and content of 0.77 g/(L·h) and 0.95 g/g dry cell weight, respectively.

On Aerial Robots with Grasping and Perching Capabilities: A Comprehensive Review.

Over the last decade, there has been an increased interest in developing aerial robotic platforms that exhibit grasping and perching capabilities not only within the research community but also in companies across different industry sectors. Aerial robots range from standard multicopter vehicles/drones, to autonomous helicopters, and fixed-wing or hybrid devices. Such devices rely on a range of different solutions for achieving grasping and perching. These solutions can be classified as: 1) simple gripper systems, 2) arm-gripper systems, 3) tethered gripping mechanisms, 4) reconfigurable robot frames, 5) adhesion solutions, and 6) embedment solutions. Grasping and perching are two crucial capabilities that allow aerial robots to interact with the environment and execute a plethora of complex tasks, facilitating new applications that range from autonomous package delivery and search and rescue to autonomous inspection of dangerous or remote environments. In this review paper, we present the state-of-the-art in aerial grasping and perching mechanisms and we provide a comprehensive comparison of their characteristics. Furthermore, we analyze these mechanisms by comparing the advantages and disadvantages of the proposed technologies and we summarize the significant achievements in these two research topics. Finally, we conclude the review by suggesting a series of potential future research directions that we believe that are promising.

Sensitivity of ASPP and P-gp to neoadjuvant chemotherapy combined with gene therapy in locally advanced cervical cancer.

PURPOSE: To evaluate the sensitivity of apoptosis-stimulating protein of p53 (ASPP) and P-glycoprotein (P-gp) to the recombinant human adenovirus-p53 (rAd-p53) combined with neoadjuvant chemotherapy for locally advanced cervical cancer and their predictive value for efficacy. METHODS: 80 patients with histopathologically diagnosed locally advanced cervical cancer (stage Ib2-IIa2) and operative treatment from February 2009 to June 2013 were enrolled and randomly divided into 3 groups: radical hysterectomy group (RH group, n=30), simple intravenous chemotherapy with cisplatin paclitaxel group (TP group, n=30) and gene therapy + intravenous chemotherapy group with cisplatin+paclitaxel (rAd-p53 + TP group, n=20). Patients in the RH group were directly operated, those in the TP group were treated with TP chemotherapy for 2 courses, and those in the rAd-p53 + TP group were treated with the same chemotherapy regimen and the rAd-p53 agent (1×1012 virus particles) was injected for the first time 3 days after chemotherapy for 3 times. The changes in tumor volume, adverse reactions and survival of patients in the TP group and rAd-p53 + TP group, and the levels of p53, ASPP2, inhibitory member of the ASPP family (iASPP) and P-gp in postoperative tumor tissues in the 3 groups were evaluated. RESULTS: The efficacy was evaluated for all patients in the TP group and rAd-p53 + TP group 3 weeks after chemotherapy. The tumor was reduced by 10.90±2.62 cm2 after medication in the TP group and 15.25±4.01 cm2 after medication in rAd-p53 + TP group, and the difference was statistically significant. The response rate [complete response (CR) + partial response (PR)] was 76.7% in the TP group and 95% in the rAd-p53 + TP group, with statistically significant difference. The positive expression rate of p53 protein showed a gradual decreasing trend in the RH group, TP group and rAd-p53 + TP group, with statistically significant difference (p<0.05). The positive expression rate of ASPP2 displayed a gradual increasing trend in the RH group, TP group and rAd-p53 + TP group, without statistically significant difference (p>0.05). The positive expression rate of iASPP was different and gradually declined in the RH group, TP group and rAd-p53 + TP group (p<0.05). Moreover, the positive expression rate of P-gp was also different among the 3 groups, and it was increased in the TP group, while it was decreased in the RH and rAd-p53 + TP group (p<0.05). CONCLUSION: The intratumor injection of rAd-p53 into patients with cervical cancer is safe and effective, showing a new way in the gene therapy of cervical cancer. Both ASPP and P-gp may be potential biomarkers in the treatment of cervical cancer.

Neuromorphic photonics with electro-absorption modulators.

Photonic neural networks benefit from both the high-channel capacity and the wave nature of light acting as an effective weighting mechanism through linear optics. Incorporating a nonlinear activation function by using active integrated photonic components allows neural networks with multiple layers to be built monolithically, eliminating the need for energy and latency costs due to external conversion. Interferometer-based modulators, while popular in communications, have been shown to require more area than absorption-based modulators, resulting in a reduced neural network density. Here, we develop a model for absorption modulators in an electro-optic fully connected neural network, including noise, and compare the network's performance with the activation functions produced intrinsically by five types of absorption modulators. Our results show the quantum well absorption modulator-based electro-optic neuron has the best performance allowing for 96% prediction accuracy with 1.7×10-12 J/MAC excluding laser power when performing MNIST classification in a 2 hidden layer feed-forward photonic neural network.