Polymeric nanoparticles (PNPs) for oral delivery of insulin.

Successful oral insulin administration can considerably enhance the quality of life (QOL) of diabetes patients who must frequently take insulin injections. Oral insulin administration, on the other hand, is seriously hampered by gastrointestinal enzymes, wide pH range, mucus and mucosal layers, which limit insulin oral bioavailability to ≤ 2%. Therefore, a large number of technological solutions have been proposed to increase the oral bioavailability of insulin, in which polymeric nanoparticles (PNPs) are highly promising for oral insulin delivery. The recently published research articles chosen for this review are based on applications of PNPs with strong future potential in oral insulin delivery, and do not cover all related work. In this review, we will summarize the controlled release mechanisms of oral insulin delivery, latest oral insulin delivery applications of PNPs nanocarrier, challenges and prospect. This review will serve as a guide to the future investigators who wish to engineer and study PNPs as oral insulin delivery systems.

Modulating the gut microbiota is involved in the effect of low-molecular-weight Glycyrrhiza polysaccharide on immune function.

Low molecular weight (6.5 kDa) Glycyrrhiza polysaccharide (GP) exhibits good immunomodulatory activity, however, the mechanism underlying GP-mediated regulation of immunity and gut microbiota remains unclear. In this study, we aimed to reveal the mechanisms underlying GP-mediated regulation of immunity and gut microbiota using cyclophosphamide (CTX)-induced immunosuppressed and intestinal mucosal injury models. GP reversed CTX-induced intestinal structural damage and increased the number of goblet cells, CD4+, CD8+ T lymphocytes, and mucin content, particularly by maintaining the balance of helper T lymphocyte 1/helper T lymphocyte 2 (Th1/Th2). Moreover, GP alleviated immunosuppression by down-regulating extracellular regulated protein kinases/p38/nuclear factor kappa-Bp50 pathways and increasing short-chain fatty acids level and secretion of cytokines, including interferon-γ, interleukin (IL)-4, IL-2, IL-10, IL-22, and transforming growth factor-ß3 and immunoglobulin (Ig) M, IgG and secretory immunoglobulin A. GP treatment increased the total species and diversity of the gut microbiota. Microbiota analysis showed that GP promoted the proliferation of beneficial bacteria, including Muribaculaceae_unclassified, Alistipes, Lachnospiraceae_NK4A136_group, Ligilactobacillus, and Clostridia_vadinBB60_group, and reduced the abundance of Proteobacteria and CTX-derived bacteria (Clostridiales_unclassified, Candidatus_Arthromitus, Firmicutes_unclassified, and Clostridium). The studies of fecal microbiota transplantation and the pseudo-aseptic model conformed that the gut microbiota is crucial in GP-mediated immunity regulation. GP shows great potential as an immune enhancer and a natural medicine for treating intestinal inflammatory diseases.

A promising and highly sensitive electrochemical platform for the detection of fentanyl and alfentanil in human serum.

A novel electrochemical method has been developed, based on a covalent organic framework (COF) and reduced graphene oxide (rGO), to detect fentanyl and alfentanil. COF nanomaterials with chrysanthemum morphology obtained by solvothermal reaction contain rich active sites for electrochemical catalytic reaction, thus improving the detection performance of the designed sensor. Reduced graphene oxide improves the sensor's sensitivity due to enhanced electron transfer. Under optimized experimental conditions, the fabricated electrode presents a linear range of 0.02 to 7.26 µM for alfentanil and 0.1 to 6.54 µM for fentanyl, with detection limits of 6.7 nM and 33 nM, respectively. In addition, the sensor possesses excellent selectivity, outstanding reproducibility, and acceptable stability. The proposed sensor is feasible for the reliable monitoring of fentanyl and alfentanil in human serum samples, with acceptable reliability and high potential in real-world applications. Finally, the electrochemical characteristic fingerprint of fentanyl is investigated by studying the electrochemical behavior of alfentanil and fentanyl on the electrode surface.

Characterization and heterologous expression of plasmalogen synthase MeHAD from Megasphaera elsdenii.

Plasmalogens (Pls) are vinyl-ether bond-containing glycerophospholipids or glycosyl diradyl glycerols, and are of great importance in the physiological functions and stability of cell membrane. Here, we identified and characterized that the plasmalogen synthase MeHAD from anaerobic Megasphaera elsdenii was responsible for vinyl-ether bond formation. Different from the 2-hydroxyacyl-CoA dehydratase (HAD) family plasmalogen synthase PlsA-PlsR which are encoded by two genes in Clostridium perfringens, the HAD homolog (MeHAD) encoded by a single gene MELS_0169 was found in M. elsdenii. By heterologous expression of the MeHAD gene into a nonplasmalogen-producing Escherichia coli strain, the expressed MeHAD was found to be located in the cell membrane region. Plasmalogens were detected in the recombinant strain using GC-MS and LC-MS, demonstrating that MeHAD was the key enzyme for plasmalogen synthesis. Moreover, the synthesized plasmalogens could enhance the oxidative stress-resistance and osmotic pressure-resistance of the recombinant strain, probably due to the ROS scavenging and decreased membrane permeability by the plasmalogens, respectively. The four-cysteine (Cys125, Cys164, Cys445 and Cys484) site-mutant of MeHAD, which were predicted binding to the [4Fe-4S] cluster, was unable to synthesize plasmalogens, indicating that the cysteines are important for the catalytic activity of MeHAD. Our results revealed the single gene encoded plasmalogen synthase in M. elsdenii and established a recombinant E. coli strain with plasmalogen production potential.

Development of Terminator-Promoter Bifunctional Elements for Application in Saccharomyces cerevisiae Pathway Engineering.

The construction of a genetic circuit requires the substitution and redesign of different promoters and terminators. The assembly efficiency of exogenous pathways will also decrease significantly when the number of regulatory elements and genes is increased. We speculated that a novel bifunctional element with promoter and terminator functions could be created via the fusion of a termination signal with a promoter sequence. In this study, the elements from a Saccharomyces cerevisiae promoter and terminator were employed to design a synthetic bifunctional element. The promoter strength of the synthetic element is apparently regulated through a spacer sequence and an upstream activating sequence (UAS) with a ~5-fold increase, and the terminator strength could be finely regulated by the efficiency element, with a ~5-fold increase. Furthermore, the use of a TATA box-like sequence resulted in the adequate execution of both functions of the TATA box and the efficiency element. By regulating the TATA box-like sequence, UAS, and spacer sequence, the strengths of the promoter-like and terminator-like bifunctional elements were optimally fine-tuned with ~8-fold and ~7-fold increases, respectively. The application of bifunctional elements in the lycopene biosynthetic pathway showed an improved pathway assembly efficiency and higher lycopene yield. The designed bifunctional elements effectively simplified pathway construction and can serve as a useful toolbox for yeast synthetic biology.

An electrochemical sensor for direct and sensitive detection of ketamine.

Ketamine is an organic drug with weak electrochemical activity, which makes it difficult to directly detect by electrochemical methods. Herein, an electrochemical sensor, with excellent detection sensitivity, is proposed for direct detection of ketamine based on a weakly conductive poly-L-cysteine molecularly imprinted membrane. Poly-L-cysteine molecularly imprinted membrane sensor (poly-L-Cys-KT-MIM/GCE) is obtained using L-cysteine as a functional monomer and ketamine as a template molecule based on electropolymerization. The green and highly active cysteine is selected as a functional monomer during electropolymerization, which cannot only achieve specific recognition but also improve detection sensitivity. Furthermore, the oxidation mechanism and fingerprint of ketamine on the electrode surface are established by analyzing the corresponding oxidation products using high/resolution mass spectrometry, which will help to promote the application of electrochemistry in the rapid detection of drugs. Under optimal conditions, the as-designed sensor demonstrated a linear response to ketamine within the range of 5.0 × 10-7 to 2.0 × 10-5 mol L-1 and a detection limit of 1.6 × 10-7 mol L-1. The proposed method exhibited excellent performance from the viewpoints of selectivity, sensitivity and stability. Notably, the sensor rendered excellent reliability and could be used for the detection of target analytes in hair and urine samples with high recovery rates.

Enhanced Thermoelectric Properties of p-Type Bi0.5Sb1.5Te3-Cu8GeSe6 Composite Materials.

Bismuth-telluride-based thermoelectric materials have been applied in active room-temperature cooling, but the mediocre ZT value of ∼1.0 limits the thermoelectric (TE) device's conversion efficiency and determines its application. In this work, we show the obviously improved thermoelectric properties of p-type Bi0.5Sb1.5Te3 by the Cu8GeSe6 composite. The addition of Cu8GeSe6 effectively boosts the carrier concentration and thus limits the bipolar thermal conductivity as the temperature is elevated. With the Cu8GeSe6 content of 0.08 wt %, the hole concentration reaches 5.0 × 1019 cm-3 and the corresponding carrier mobility is over 160 cm2 V-1 s-1, resulting in an optimized power factor of over 42 µW cm-1 K-2 at 300 K. Moreover, the Cu8GeSe6 composite introduces multiple phonon-scattering centers by increasing dislocations and element and strain field inhomogeneities, which reduce the thermal conductivity consisting of a lattice contribution and a bipolar contribution to 0.51 W m-1 K-1 at 350 K. As a consequence, the peak ZT of the Bi0.5Sb1.5Te3-0.08 wt % Cu8GeSe6 composite reaches 1.30 at 375 K and the average ZT between 300 and 500 K is improved to 1.13. A thermoelectric module comprised of this composite and commercial Bi2Te2.5Se0.5 exhibits a conversion efficiency of 5.3% with a temperature difference of 250 K, demonstrating the promising applications in low-grade energy recovery.

Highly sensitive triethylamine gas sensor by Pt-loaded p-n heterojunction Co3O4/WO3.

Triethylamine (TEA) exists widely in production and life and is extremely volatile, which seriously endangers human health. It is required to develop high-performance TEA sensors to protect human health. We fabricated Pt-Co3O4/WO3based on our previous work, and the performance was tested against volatile organic compounds. Compared with the previous work, its operating temperature was greatly reduced from 240 °C to 180 °C. The response value of Pt-Co3O4/WO3was increased from 1101 to 1532 for 10 ppm TEA with good selectivity. These results show a significant step toward practical use of the Pt-Co3O4/WO3sensor.

Natural Polysaccharide-Based Nanodrug Delivery Systems for Treatment of Diabetes.

In recent years, natural polysaccharides have been considered as the ideal candidates for novel drug delivery systems because of their good biocompatibility, biodegradation, low immunogenicity, renewable source and easy modification. These natural polymers are widely used in the designing of nanocarriers, which possess wide applications in therapeutics, diagnostics, delivery and protection of bioactive compounds or drugs. A great deal of studies could be focused on developing polysaccharide nanoparticles and promoting their application in various fields, especially in biomedicine. In this review, a variety of polysaccharide-based nanocarriers were introduced, including nanoliposomes, nanoparticles, nanomicelles, nanoemulsions and nanohydrogels, focusing on the latest research progress of these nanocarriers in the treatment of diabetes and the possible strategies for further study of polysaccharide nanocarriers.

Carotenoid Biosynthesis: Genome-Wide Profiling, Pathway Identification in Rhodotorula glutinis X-20, and High-Level Production.

Rhodotorula glutinis, as a member of the family Sporidiobolaceae, is of great value in the field of biotechnology. However, the evolutionary relationship of R. glutinis X-20 with Rhodosporidiobolus, Sporobolomyces, and Rhodotorula are not well understood, and its metabolic pathways such as carotenoid biosynthesis are not well resolved. Here, genome sequencing and comparative genome techniques were employed to improve the understanding of R. glutinis X-20. Phytoene desaturase (crtI) and 15-cis-phytoene synthase/lycopene beta-cyclase (crtYB), key enzymes in carotenoid pathway from R. glutinis X-20 were more efficiently expressed in S. cerevisiae INVSc1 than in S. cerevisiae CEN.PK2-1C. High yielding engineered strains were obtained by using synthetic biology technology constructing carotenoid pathway in S. cerevisiae and optimizing the precursor supply after fed-batch fermentation with palmitic acid supplementation. Genome sequencing analysis and metabolite identification has enhanced the understanding of evolutionary relationships and metabolic pathways in R. glutinis X-20, while heterologous construction of carotenoid pathway has facilitated its industrial application.

Microwave-prepared surface imprinted magnetic nanoparticles based electrochemical sensor for adsorption and determination of ketamine in sewage.

Detection technology for the determination of drugs, such as ketamine (KT), in sewage is of great significance in drug inspection and criminal investigation. Herein, we propose the utilization of ketamine magnetic molecularly imprinted polymers (Fe3O4@MIPs) as a target molecule identification and concentration container coupled with magnetic glassy carbon electrode (mGCE) for KT detection in sewage. Molecular simulations were employed to evaluate the most suitable monomer and ratio of functional monomer to template. Fe3O4@MIPs were prepared using microwave-assisted synthesis and possessed a "shell-core" structure with good recognition ability, superior adsorption capacity and fast kinetics toward KT. Additionally, a novel imprinted electrochemical sensor was constructed based on the magnetism of Fe3O4@MIPs for efficient monitoring of low concentrations of KT. The morphology and properties of Fe3O4@MIPs/mGCE were effectively characterized by element mapping, transmission electron microscopy, cyclic voltammetry and square wave voltammetry. KT detection was performed by square wave voltammetry within the range of 1.0 × 10-12 and 4.0 × 10-4 mol L-1, and the limit of detection was 8.0 × 10-13 mol L-1. Furthermore, Fe3O4@MIPs/mGCE was successfully tested for KT determination in domestic sewage samples.

High efficient production of plant flavonoids by microbial cell factories: Challenges and opportunities.

Plant flavonoids are secondary metabolites containing a benzo-γ-pyrone structure, which are widely present in plants and have a variety of physiological and pharmacological activities. However, current flavonoid production from plant extraction or chemical synthesis does not meet the requirements of green and sustainable development. Fortunately, microbial synthesis of flavonoids has shown the potential for large-scale production with the advantages of being controllable and environmentally friendly, and a variety of microorganisms have been developed as microbial cell factories (MCFs) to synthesize plant flavonoids owing to the feasibility of genetic manipulations. However, most of MCFs have not yet been commercialized and industrialized because of the challenges posed by unbalanced metabolic flux among various pathways and conflict between cell growth and production. Here, strategies for coping with the challenges are summarized in terms of enzymes, pathways, metabolic networks, host cells. And combined with protein structure prediction, de novo protein design, artificial intelligence (AI), biocatalytic retrosynthesis, and intelligent stress resistance, it provides new insights for the high efficient production of plant flavonoids and other plant natural products in MCFs.

Pt Single Atom-Induced Activation Energy and Adsorption Enhancement for an Ultrasensitive ppb-Level Methanol Gas Sensor.

As an important organic chemical raw material, methanol is used in various industries but is harmful to human health. Developing an effective and accurate detection device for methanol is an urgent need. Herein, we demonstrate a novel gas-sensing material with a Pt single atom supported on a porous Ag-LaFeO3@ZnO core-shell sphere (Ag-LaFeO3@ZnO-Pt) with a high specific surface area (192.08 m2·g-1). Based on this, the surface activity of the Ag-LaFeO3@ZnO-Pt gas sensor is enhanced obviously, which improved the working temperature and detection limit for methanol gas. Consequently, this sensor possesses an ultrahigh sensitivity of 453.02 for 5 ppm methanol gas at a working temperature of 86 °C and maintains a high sensitivity of 21.25 even at a concentration as low as 62 ppb. The sensitivity of Ag-LaFeO3@ZnO-Pt to methanol gas is increased by 6.69 times compared with the Ag-LaFeO3@ZnO core-shell sphere (Ag-LaFeO3@ZnO). Additionally, the minimum detection limit is found to be 3.27 ppb. Detailed theoretical calculations revealed that the unoccupied 5d state of Pt single atoms increases the adsorption and activation energy of methanol and oxygen, which facilities methanol gas-sensing performance. This work will provide a novel strategy to design high-performance gas-sensing materials.

Ultrasensitive ppb-level trimethylamine gas sensor based on p-n heterojunction of Co3O4/WO3.

Trace poisonous and harmful gases in the air have been harming and affecting people's health for a long time. At present, effective and accurate detection of ppb-level harmful gas is still a bottleneck to be overcome. Herein, we report a ppb-level triethylamine (TEA) gas sensor based on p-n heterojunction of Co3O4/WO3, which is prepared with ZIF-67 as the precursor and provides Co3O4deposited tungsten oxide flower-like structure. Due to the introduction of Co3O4and the 3D flower-like structure of WO3, the Co3O4/WO3-2 gas sensor shows excellent gas sensing performance (1101 for 10 ppm at 240 °C), superb selectivity, good long-term stability and linear response for TEA concentration. Moreover, the experimental results indicate that the Co3O4/WO3-2 gas sensor also possesses a good response to 50 ppb TEA, in fact, the theoretical limit of detection is 0.6 ppb. Co3O4not only improves the efficiency of electron separation/transport, but also accelerates the oxidation rate of TEA. This method of synthesizing p-n heterojunction with ZIF as the precursor provides a new idea and method for the preparation of low detection limit gas sensors.

Sensitive detection of ketamine with an electrochemical sensor based on UV-induced polymerized molecularly imprinted membranes at graphene and MOFs modified electrode.

Ketamine is one of the most widely abused drugs in the world and poses a serious threat to human health and social stability; therefore, the ability to accurately monitor the substance in real-time is necessary. However, several problems still exists towards this goal, such as the generally low concentration of the target molecules disturbed in the complex samples that undergo analysis during criminal investigations. In this work, the sensitive and selective detection of ketamine was accomplished by molecularly imprinted electrochemical sensor. The molecularly imprinted membrane as a biomimetic recognition element was fabricated by the UV-induced polymerization of methacrylic acid (MAA) and ethylene glycol dimethacrylate (EGDMA) on a metal-organic framework/graphene nanocomposite (MOFs@G) modified screen-printed electrode. The screen printed electrode (SPE) provided good adhesion for the formation of the imprinted membranes and increased the stability of the sensor. The morphology and performance of the imprinted films were characterized in detail by scanning electron microscopy (SEM), cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), and differential pulse voltammetry (DPV). The experimental results demonstrated that the imprinted sensor had excellent sensitivity, selectivity, and long-term stability. It offered a low detection limit (4.0 × 10-11 mol L-1) and had a dynamic range from 1.0 × 10-10 mol L-1 to 4.0 × 10-5 mol L-1. Furthermore, the established method was successfully applied for the determination of ketamine in urine and saliva samples.

Improvement of selenium enrichment in Rhodotorula glutinis X-20 through combining process optimization and selenium transport.

Selenium-enriched yeast can transform toxic inorganic selenium into absorbable organic selenium, which is of great significance for human health and pharmaceutical industry. A yeast Rhodotorula glutinis X-20 we obtained before has good selenium-enriched ability, but its selenium content is still low for industrial application. In this study, strategies of process optimization and transport regulation of selenium were thus employed to further improve the cell growth and selenium enrichment. Through engineering phosphate transporters from Saccharomyces cerevisiae into R. glutinis X-20, the selenium content was increased by 21.1%. Through using mixed carbon culture (20 g L-1, glycerol: glucose 3:7), both biomass and selenium content were finally increased to 5.3 g L-1 and 5349.6 µg g-1 (cell dry weight, DWC), which were 1.14 folds and 6.77 folds compared to their original values, respectively. Our results indicate that high selenium-enrichment ability and biomass production can be achieved through combining process optimization and regulation of selenium transport.

Yeast Synthetic Terminators: Fine Regulation of Strength through Linker Sequences.

The design of improved synthetic components is an important research field in synthetic biology. The terminator, responsible for terminating gene transcription, is a necessary component for yeast gene expression. The efficiency element, the positioning element and the poly(A) site have been identified as the constituent parts necessary for the yeast terminator to perform its function. However, the functions of linker 1 (situated between the efficiency element and the positioning element) and linker 2 [between the positioning element and the poly(A) site] in the terminator are still controversial. Here, we have thus designed and synthesized a yeast synthetic terminator library incorporating random 10 bp linker 1 units. For indirect characterization of the strengths of 266 synthetic terminators with the aid of the enhanced green fluorescent protein (eGFP), their fluorescence intensity (FI) values were determined; they ranged from 2.3648 to 3.5270, thus indicating that the strength of yeast terminator can be finely adjusted by changing the linker 1 sequence. The strength increased with decreasing GC content in linker 1, with a T-rich linker 1 helping to enhance terminator strength further. Reducing the stem length can increase the gene expression in cases of weak and medium-strength terminators but decreases the gene expression of strong terminators. Deletion of linker 2 seems to have a positive effect on weak and medium-strength terminators. Construction of a lycopene biosynthesis pathway with synthetic terminators effectively regulated lycopene synthesis, thus indicating that it is highly feasible to use terminators for fine regulation of gene and pathway expression.

Simultaneously down-regulation of multiplex branch pathways using CRISPRi and fermentation optimization for enhancing ß-amyrin production in Saccharomyces cerevisiae.

The production of ß-amyrin in Saccharomyces cerevisiae is still low due to the inability of effectively regulating the endogenous metabolic pathway for competitive synthesis of ß-amyrin precursors. In this study, we focused on two branches of ß-amyrin synthetics pathway that consume ß-amyrin precursors (2,3-oxidosqualene and cytosolic acetyl-CoA) and regulated related genes (ADH1, ADH4, ADH5, ADH6, CIT2, MLS2 and ERG7). We developed a CRISPRi method by constructing a multi-gRNA plasmid to down-regulate the seven genes simultaneously, which is reported for the first time in S. cerevisiae. The average transcription inhibition efficiency of the seven genes reached as high as 75.5%. Furthermore, by optimizing the fermentation condition (including pH, inoculum size, initial glucose concentration and feed of glucose or ethanol) and increasing extracellular transportation via supplying methyl-ß-cyclodextrin, ß-amyrin concentration of engineered strain SGibSdCg increased by 44.3% compared with the parent strain SGib, achieving 156.7 mg/L which was the highest concentration of ß-amyrin reported in yeast. The one-step down-regulation of multiple genes using CRISPRi showed high efficiency and promising future in improving the yields of natural products.

Simultaneous extraction and enrichment of polyphenol and lutein from marigold (Tagetes erecta L.) flower by an enzyme-assisted ethanol/ammonium sulfate system.

Enzyme-assisted aqueous two-phase extraction (EA-ATPE) using ethanol/ammonium sulfate system was investigated for total polyphenol (TP) and lutein from marigold flowers. The important factors were investigated by single factor experiment and response surface methodology combined with Box-Behnken design to optimize the operating parameters of EA-ATPE. The maximum yields of TP and lutein were 83.56 ± 0.69 mg g-1 and 5.59 ± 0.13 mg g-1, respectively. Compared with aqueous two-phase extraction and Soxhlet extraction (SE), the extraction yield of TP by EA-ATPE is 64.91% higher and the extract of EA-ATPE has better antioxidant activity. The pretreatment effect was also researched by scanning electron microscopy. Thus, EA-ATPE is an efficient method for extracting bioactive components from plants.

Boosted Visible-Light Photodegradation of Methylene Blue by V and Co Co-Doped TiO2.

In this work, TiO2 photocatalysts, co-doped with transition metal ions vanadium (V) and cobalt (Co) ((V,Co)⁻TiO2), were synthesized by the sol⁻gel method. The synthesized photocatalysts were characterized by X-ray diffraction (XRD), transmission electron microscopy (TEM), X-ray photoelectron spectroscopy (XPS), nitrogen adsorption and desorption measurement, UV-Vis absorption and photoluminescence spectrum (PL) spectra. The results show that V and Co co-doping has significant effects on sample average crystalline grain size, absorption spectrum, recombination efficiency of photo-induced electron-hole pairs (EHPs), and photocatalytic degradation efficiency of methylene blue (MB). (V,Co)⁻TiO2 photocatalyst exhibits an obvious red shift of the absorption edge to 475 nm. Photocatalytic degradation rate of (V,Co)⁻TiO2 sample for MB in 60 min is 92.12% under a Xe lamp with a cut-off filter (λ > 400 nm), which is significantly higher than 56.55% of P25 under the same conditions. The first principles calculation results show that V and Co ions doping introduces several impurity energy levels, which can modulate the location of the valence band and conduction band. An obvious lattice distortion is produced in the meantime, resulting in the decrease in photo-generated EHP recombination. Thus, (V,Co)⁻TiO2 photocatalyst performance is significantly improved.