Gastrointestinal disturbance and effect of fecal microbiota transplantation in discharged COVID-19 patients.

BACKGROUND: To investigate the potential beneficial effect of fecal microbiota transplantation (FMT) on gastrointestinal symptoms, gut dysbiosis and immune status in discharged COVID-19 patients. CASE PRESENTATION: A total of 11 COVID-19 patients were recruited in April, 2020, about one month on average after they were discharged from the hospital. All subjects received FMT for 4 consecutive days by oral capsule administrations with 10 capsules for each day. In total, 5 out of 11 patients reported to be suffered from gastrointestinal symptoms, which were improved after FMT. After FMT, alterations of B cells were observed, which was characterized as decreased naive B cell (P = 0.012) and increased memory B cells (P = 0.001) and non-switched B cells (P = 0.012).The microbial community richness indicated by operational taxonomic units number, observed species and Chao1 estimator was marginally increased after FMT. Gut microbiome composition of discharged COVID-19 patients differed from that of the general population at both phylum and genera level, which was characterized with a lower proportion of Firmicutes (41.0%) and Actinobacteria (4.0%), higher proportion of Bacteroidetes (42.9%) and Proteobacteria (9.2%). FMT can partially restore the gut dysbiosis by increasing the relative abundance of Actinobacteria (15.0%) and reducing Proteobacteria (2.8%) at the phylum level. At the genera level, Bifidobacterium and Faecalibacterium had significantly increased after FMT. CONCLUSIONS: After FMT, altered peripheral lymphocyte subset, restored gut microbiota and alleviated gastrointestinal disorders were observe, suggesting that FMT may serve as a potential therapeutic and rehabilitative intervention for the COVID-19.

An integrated microfluidics for assessing the anti-aging effect of caffeic acid phenethylester in Caenorhabditis elegans.

Aging is a fundamental and fascinating process. Anti-aging research tried to find the mysteries about the human lifespan. To investigate the longevity-extending role of caffeic acid phenethylester (CAPE) and reveal the possible regulation mechanism, the long-term cultivation under well-defined environments, real-time monitoring, and live imaging is highly desired. In this paper, a well-designed microfluidic device was proposed to analyze the anti-aging effect of CAPE in Caenorhabditis elegans. With the combined use of multiple functional units, including micro-pillar, worm responder, a branching network of distribution channels, and microchambers, the longitudinal measurements of the exact number of worms throughout the whole lifespans is possible. Meanwhile, the brief cooling function of temperature-controllable system can achieve temporary and repeated immobilization of nematodes for fluorescence imaging. Our research data showed that CAPE can increase the survival of worms under normal and stress condition, including heat stress and paraquat-induced oxidative stress. The further studies revealed the anti-aging mechanism of CAPE. This proposed strategy and device would be a useful platform to facilitate future anti-aging studies and the finding of new lead compounds.

Advances in the research on the targets of anti-malaria actions of artemisinin.

Malaria has been a global epidemic health threat since ancient times. It still claims roughly half a million lives every year in this century. Artemisinin and its derivatives, are frontline antimalarial drugs known for their efficacy and low toxicity. After decades of wide use, artemisinins remain our bulwark against malaria. Here, we review decades of efforts that aim to understand the mechanism of action (MOA) of artemisinins, which help explain the specificity and potency of this anti-malarial drug. We summarize the methods and approaches employed to unravel the MOA of artemisinin over the last three decades, showing how the development of advanced techniques can help provide mechanistic insights and resolve some long-standing questions in the field of artemisinin research. We also provide examples to illustrate how to better repurpose artemisinins for anti-cancer therapies by leveraging on MOA. These examples point out a practical direction to engineer artemisinin for broader applications beyond malaria.

Comparative toxicity of lead (Pb(2+)), copper (Cu(2+)), and mixtures of lead and copper to zebrafish embryos on a microfluidic chip.

Investigations were conducted to determine acute effects of Pb(2+) and Cu(2+) presented individually and collectively on zebrafish embryos. Aquatic safety testing requires a cheap, fast, and highly efficient platform for real-time evaluation of single and mixture of metal toxicity. In this study, we have developed a microfluidic system for phenotype-based evaluation of toxic effects of Pb(2+) and Cu(2+) using zebrafish (Danio rerio) embryos. The microfluidic chip is composed of a disc-shaped concentration gradient generator and 24 culture chambers, which can generate one blank solution, seven mixture concentrations, and eight single concentrations for each metal solution, thus enabling the assessment of zebrafish embryos. To test the accuracy of this new chip platform, we have examined the toxicity and teratogenicity of Pb(2+) and Cu(2+) on embryos. The individual and combined impact of Pb(2+) and Cu(2+) on zebrafish embryonic development was quantitatively assessed by recording a series of physiological indicators, such as spontaneous motion at 22 hours post fertilization (hpf), mortality at 24 hpf, heartbeat and body length at 96 hpf, etc. It was found that Pb(2+) or Cu(2+) could induce deformity and cardiovascular toxicity in zebrafish embryos and the mixture could induce more severe toxicity. This chip is a multiplexed testing apparatus that allows for the examination of toxicity and teratogenicity for substances and it also can be used as a potentially cost-effective and rapid aquatic safety assessment tool.

Microfluidic platform integrated with worm-counting setup for assessing manganese toxicity.

We reported a new microfluidic system integrated with worm responders for evaluating the environmental manganese toxicity. The micro device consists of worm loading units, worm observing chambers, and a radial concentration gradient generator (CGG). Eight T-shape worm loading units of the micro device were used to load the exact number of worms into the corresponding eight chambers with the assistance of worm responders and doorsills. The worm responder, as a key component, was employed for performing automated worm-counting assay through electric impedance sensing. This label-free and non-invasive worm-counting technique was applied to the microsystem for the first time. In addition, the disk-shaped CGG can generate a range of stepwise concentrations of the appointed chemical automatically and simultaneously. Due to the scalable architecture of radial CGG, it has the potential to increase the throughput of the assay. Dopaminergic (DAergic) neurotoxicity of manganese on C. elegans was quantitatively assessed via the observation of green fluorescence protein-tagged DAergic neurons of the strain BZ555 on-chip. In addition, oxidative stress triggered by manganese was evaluated by the quantitative fluorescence intensity of the strain CL2166. By scoring the survival ratio and stroke frequency of worms, we characterized the dose- and time-dependent mobility defects of the manganese-exposed worms. Furthermore, we applied the microsystem to investigate the effect of natural antioxidants to protect manganese-induced toxicity.

Zebrafish on a chip: a novel platform for real-time monitoring of drug-induced developmental toxicity.

Pharmaceutical safety testing requires a cheap, fast and highly efficient platform for real-time evaluation of drug toxicity and secondary effects. In this study, we have developed a microfluidic system for phenotype-based evaluation of toxic and teratogenic effects of drugs using zebrafish (Danio rerio) embryos and larvae as the model organism. The microfluidic chip is composed of two independent functional units, enabling the assessment of zebrafish embryos and larvae. Each unit consists of a fluidic concentration gradient generator and a row of seven culture chambers to accommodate zebrafish. To test the accuracy of this new chip platform, we examined the toxicity and teratogenicity of an anti-asthmatic agent-aminophylline (Apl) on 210 embryos and 210 larvae (10 individuals per chamber). The effect of Apl on zebrafish embryonic development was quantitatively assessed by recording a series of physiological indicators such as heart rate, survival rate, body length and hatch rate. Most importantly, a new index called clonic convulsion rate, combined with mortality was used to evaluate the toxicities of Apl on zebrafish larvae. We found that Apl can induce deformity and cardiovascular toxicity in both zebrafish embryos and larvae. This microdevice is a multiplexed testing apparatus that allows for the examination of indexes beyond toxicity and teratogenicity at the sub-organ and cellular levels and provides a potentially cost-effective and rapid pharmaceutical safety assessment tool.

A two-electrode system-based electrochemiluminescence detection for microfluidic capillary electrophoresis and its application in pharmaceutical analysis.

A two-electrode configuration powered by batteries was designed for a microchip capillary electrophoresis-electrochemiluminescence system. A home-made working electrode for end-column mode detection and wall-jet configuration was made up of a platinum wire (0.3 mm diameter) and a quartz capillary (320 µm internal diameter). The platinum wire served as a pseudoreference electrode. The configuration of the detection power supply comprised two D-size batteries (connected in series), a switch, and an adjustable resistor. The microchip consisted of two layers: the bottom layer was a glass sheet containing injection and separation channels; the upper layer was polydimethylsiloxane block. In order to reduce the loss of electrochemiluminescence signal, a coverslip (0.17 mm thickness) was used as the floor of the detection reservoir. The performance of the system was demonstrated by separation and detection of atropine, anisodamine and proline. The linear response for proline ranged from 5 µM to 100 µM (r = 0.9968), and the limit of detection was 1.0 µM (S/N = 3). The system was further applied to the measurement of atropine in atropine sulfate injection solutions with the limit of detection 2.3 µM. This new system is a potential tool in pharmaceutical analysis.

Differential pulsed amperometry coupled to microchip capillary electrophoresis.

In this work, we describe a novel electrochemical detection method, differential pulsed amperometry (DPA) on microchip capillary electrophoresis (MCE). In a pulse period, a sequential two-step sampling is executed at two different potentials (E1 and E2). Differential current signal of the duplex sampling events is recorded that functions as time domain. The performance of this detection scheme was evaluated by separating and detecting three model analytes including tyramine (Tym), tryptophan (Trp), and p-aminobenzoic acid (PABA). Multiple parameters that would affect electrochemical response and peak shape, such as sampling potential, sampling time, and electrode cleaning time, were investigated. This pulse technique exhibits better sensitivity over constant potential amperometry (CPA), nearly equal to triple pulsed amperometry (TPA). More importantly, DPA can generate more stable baseline than TPA, primarily due to the background subtraction through the two-step sampling, which is beneficial to further improve analytical sensitivity. In the optimal condition, the limits of detection for Tym, Trp and PABA, were down to 0.27µM, 0.32µM and 1.1µM, respectively. DPA detection opens up a new avenue for microchip electrochemistry, and can be virtually extended to other fluid analysis techniques.

Development of a microchip-pulsed electrochemical method for rapid determination of L-DOPA and tyrosine in Mucuna pruriens.

L-3,4-dihydroxyphenylalanine (L-DOPA) is a well-recognized therapeutic compound to Parkinson's disease. Tyrosine is a precursor for the biosynthesis of L-DOPA, both of which are widely found in traditional medicinal material, Mucuna pruriens. In this paper, we described a validated novel analytical method based on microchip capillary electrophoresis with pulsed electrochemical detection for the simultaneous measurement of L-DOPA and tyrosine in M. pruriens. This protocol adopted end-channel amperometric detection using platinum disk electrode on a homemade glass/polydimethylsiloxane electrophoresis microchip. The background buffer consisted of 10 mM borate (pH 9.5) and 0.02 mM cetyltrimethylammonium bromide, which can produce an effective resolution for the two analytes. In the optimal condition, sufficient electrophoretic separation and sensitive detection for the target analytes can be realized within 60 s. Both tyrosine and L-DOPA yielded linear response in the concentration range of 5.0-400 µM (R(2) > 0.99), and the LOD were 0.79 and 1.1 µM, respectively. The accuracy and precision of the established method were favorable. The present method shows several merits such as facile apparatus, high speed, low cost and minimal pollution, and provides a means for the pharmacologically active ingredients assay in M. pruriens.

catena-Poly[[tetra-aqua-cobalt(II)]-µ-2,2'-dihydroxy-5,5'-diazenediyldibenzoato].

In the title compound, [Co(C(14)H(8)N(2)O(6))(H(2)O)(4)](n), each 5,5'-diazenediylbis(2-hydroxy-benzoato) ligand acts as a dicarboxyl-ate bridge, leading to the formation of polymeric chains running in the [10] direction. The Co atom is hexa-coordinated in a distorted octa-hedral geometry by six O atoms [Co-O = 2.039 (4)-2.115 (4) Å] from two ligands and four water mol-ecules. Inter-molecular O-H⋯O and O-H⋯N hydrogen bonds build up a three-dimensional supra-molecular structure.

2,2'-Dihydroxy-3,3'-[(1E,1'E)-hydrazine-1,2-diylidenedimethylidyne]dibenzoic acid N,N-dimethylformamide disolvate.

The title compound, C(16)H(12)N(2)O(6)·2C(3)H(7)NO, lies across a crystallographic inversion centre which is situated at the midpoint of the central N-N bond. The substitution at the C=N bond adopts a trans configuration and it is essentially coplanar with the benzene ring [N-C-C-C torsion angles = -173.9 (4) and 6.4 (6)°]. All torsion angles involving non-H atoms are close to 180°. Intra-molecular O-H⋯O and weak C-H⋯O hydrogen bonds form S(6) and S(5) ring motifs, respectively, while inter-molecular O-H⋯O and weak C-H⋯O hydrogen bonds connect the Schiff base mol-ecule to solvent dimethyl-formamide mol-ecules.