Metabolic Labeling and Digital Microfluidic Single-Cell Sequencing for Single Bacterial Genotypic-Phenotypic Analysis.

Accurate assessment of phenotypic and genotypic characteristics of bacteria can facilitate comprehensive cataloguing of all the resistance factors for better understanding of antibiotic resistance. However, current methods primarily focus on individual phenotypic or genotypic profiles across different colonies. Here, a Digital microfluidic-based automated assay for whole-genome sequencing of single-antibiotic-resistant bacteria is reported, enabling Genotypic and Phenotypic Analysis of antibiotic-resistant strains (Digital-GPA). Digital-GPA can efficiently isolate and sequence antibiotic-resistant bacteria illuminated by fluorescent D-amino acid (FDAA)-labeling, producing high-quality single-cell amplified genomes (SAGs). This enables identifications of both minor and major mutations, pinpointing substrains with distinctive resistance mechanisms. Digital-GPA can directly process clinical samples to detect and sequence resistant pathogens without bacterial culture, subsequently provide genetic profiles of antibiotic susceptibility, promising to expedite the analysis of hard-to-culture or slow-growing bacteria. Overall, Digital-GPA opens a new avenue for antibiotic resistance analysis by providing accurate and comprehensive molecular profiles of antibiotic resistance at single-cell resolution.

Copper-Nanoparticle-Induced Neurotoxic Effect and Oxidative Stress in the Early Developmental Stage of Zebrafish (Danio rerio).

Copper nanoparticles (CuNPs) are extensively used in electronics, cosmetics, fungicides, and various other fields due to their distinctive qualities. However, this widespread usage can contribute to environmental contamination and heightened health risks for living organisms. Despite their prevalent use, the ecological impacts and biosafety of CuNPs remain inadequately understood. The present study aims to delve into the potential toxic effects of CuNPs on zebrafish (Danio rerio) embryos, focusing on multiple indexes such as embryonic development, neurotoxicity, oxidative stress, and inflammatory response. The results revealed a notable increase in the death rate and deformity rate, alongside varying degrees of decrease in hatching rate and heart rate following CuNPs exposure. Particularly, the frequency of spontaneous tail coiling significantly declined under exposure to CuNPs at concentrations of 500 µg/L. Furthermore, CuNPs exposure induced alterations in the transcriptional expression of GABA signaling pathway-related genes (gabra1, gad, abat, and gat1), indicating potential impacts on GABA synthesis, release, catabolism, recovery, and receptor binding. Additionally, CuNPs triggered oxidative stress, evidenced by disruption in superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) activities, along with elevated malondialdehyde (MDA) levels. This oxidative stress subsequently led to a proinflammatory cascade, as demonstrated by the increased transcriptional expression of inflammatory markers (il-1ß, tnf-α, il-6, and il-8). Comparative analysis with copper ion (provided as CuCl2) exposure highlighted more significant changes in most indexes with CuCl2, indicating greater toxicity compared to CuNPs at equivalent concentrations. In conclusion, these findings provide valuable insights into the toxic effects of CuNPs on zebrafish embryo development and neurotransmitter conduction. Furthermore, they present technical methodologies for assessing environmental and health risks associated with CuNPs, contributing to a better understanding of their biosafety and ecological impact.

Diamino-functionalized metal-organic framework for selective capture of gold ions.

Adsorptive recovery of valuable gold (Au) ions from wastes is vital but still challenged, especially regarding adsorption capacity and selectivity. A novel M - 3,5-DABA metal-organic framework (MOF) adsorbent was prepared via anchoring 3,5-diaminobenzoic acid (3,5-DABA) molecule in the MOF-808 matrix. Benefiting from the positive charge property, dense amino groups (3.2 mmol g-1) and high porosity, the adsorption capacity of M - 3,5-DABA reaches 1391.5 mg g-1 (pH = 2.5) and adsorption equilibrium is attained in 5 min. This amino-based material shows excellent selectivity towards various metal ions, evading the poor selectivity problem of classical thiol groups (e.g. for Ag+, Cu2+, Pb2+ and Hg2+ ions). In addition, the regeneration was easily achieved via using a hydrochloric acid-thiourea eluent. Experimental analysis and density functional theory (DFT) calculation show the amino group works as a reductant for Au(III) ions and meanwhile acts as an active site for adsorbing Au(III) ions together with the µ-OH group. Thus, M - 3,5-DABA can act as a potential adsorbent for Au(III) ions, and our work offers a viable strategy to construct novel MOF-based adsorbents.

Molecular Dynamics Simulation of the Rejuvenation Performance of Waste Cooking Oil with High Acid Value on Aged Asphalt.

Waste cooking oil's (WCO's) potential as a rejuvenator of aged asphalt has received attention in recent years, with the acid value of WCO affecting its rejuvenation effect. This study explored the rejuvenation effect of WCO with a high acid value on aged asphalt by using molecular dynamics simulation. First, the representative molecules of WCO with a high acid value and asphalt were determined. The rejuvenation effect of WCO on aged asphalt was analyzed by adding different contents of WCO to an aged asphalt model. The effect of WCO on the thermodynamic properties of the aged asphalt was analyzed. The results show that WCO can restore the thermodynamic properties of aged asphalt binder to a certain extent. Regarding the microstructure of rejuvenated asphalt, WCO molecules dispersed around asphaltenes weakened the latter's aggregation and improved the colloidal structure of the aged asphalt. In terms of interface adhesion properties, WCO can improve the adhesion properties between asphalt binder and SiO2, but it has limited influence on water sensitivity. The results allowed us to comprehensively evaluate the rejuvenation effect of WCO with a high acid value on aged asphalt and to explore its rejuvenation mechanism.

A young woman with Mycobacterium chelonae complex infection caused by facial stem cell injection.

In outpatient settings, Mycobacterium chelonae complex infection brought on by cosmetic injections are rather uncommon. We came across a case of infection brought on by a commercial stem cell injection.

Recent advances in exosomal non-coding RNA-based therapeutic approaches for photoaging.

BACKGROUND: Photoaging is a degenerative biological process that affects the quality of life. It is caused by environmental factors including ultraviolet radiation (UVR), deep skin burns, smoking, active oxygen, chemical substances, and trauma. Among them, UVR plays a vital role in the aging process. AIM: With the continuous development of modern medicine, clinical researchers have investigated novel approaches to treat aging. In particular, mesenchymal stem cells (MSCs), non-coding RNAs are involved in various physiological processes have broad clinical application as they have the advantages of convenient samples, abundant sources, and avoidable ethical issues. METHODS: This article reviews research progress on five types of stem cell, exosomes, non-coding RNA in the context of photoaging treatment: adipose-derived stem cell, human umbilical cord MSCs, epidermal progenitor cells, keratinocyte stem cells, and hair follicle stem cells (HFSCs). It also includes stem cell related exosomes and their non-coding RNA research. RESULTS: The results have clinical guiding significance for prevention and control of the onset and development of photoaging. It is found that stem cells secrete cytokines, cell growth factors, non-coding RNA, exosomes and proteins to repair aging skin tissues and achieve skin rejuvenation. In particular, stem cell exosomes and non-coding RNA are found to have significant research potential, as they possess the benefits of their source cells without the disadvantages which include immune rejection and granuloma formation.

Acquired syphilis with flat condyloma in a 3 year-old girl: A case report.

The incidence of syphilis in young children is very low, with acquired syphilis exceptionally rare. A 3 year-old girl presented to our service with a reddish-brown rash on the external genitals and perianal area. The rash had been apparent for a period of 1 week. The girl was asymptomatic and had no history of sexual contact. Syphilis was suspected and the Treponema pallidum particle agglutination test (TPPA) was found to be positive, as well as the rapid plasma regin ring card test (RPR), with a titer of 1:64. The girl was diagnosed with toddler acquired secondary syphilis. The girl was cured after three treatments with penicillin. This report points out the need for clinicians to be aware of nonsexually transmitted syphilis, acquired in daily life by children without a history of sexual contact.

The Molecular Model of Organic Matter in Coal-Measure Shale: Structure Construction and Evaluation Based on Experimental Characterization.

To investigate the molecular structure and micropore structure of organic matters in coal-measure shale, the black shale samples of the Shanxi formation were collected from Xishan Coalfield, Taiyuan, and a hybrid experimental-simulation method was used for realistic macromolecular models of organic matter (OM). Four experimental techniques were used to determine the structural information of OM, including elemental analysis, state 13C nuclear magnetic resonance (13CNMR), X-ray photoelectron spectroscopy (XPS), and Fourier transform infrared spectroscopy (FTIR). With structural parameters, two-dimensional (2D) average molecular models of OM were established as C177H160O8N2S with a molar weight of 2474, which agreed well with the experimental 13C-NMR spectra. A realistic three-dimensional (3D) OM macromolecular model was also reconstructed, containing 20 2D molecules with a density of 1.41 g/cm3. To determine the connectivity and spatial disposition of the OM pores, focused ion beam microscope (FIB-SEM) and transmission electron micrographs (TEM) were utilized. The 3D OM pores models were developed. The results show that whether the OM pores varied from 20 to 350 nm as obtained from FIB-SEM images or less than 10 nm as observed in the TEM images, both were of poor connectivity. However, the ultra-micro pores from the 3D OM macromolecular model varied from 3Å to 10 Å and showed certain connectivity, which may be the main channel of diffusion. Furthermore, with the pressure increased, the methane adsorption capacity of the 3D OM model increased with a maximum value of 103 cm3/g at 7 MPa, indicating that OM pores less than 1 nm have a huge methane adsorption capacity. Therefore, our work provides an analysis method that is a powerful and superior tool in further research on gas migration.

Tuning Lignite Structure via Hydromodification To Promote the Formation of Coal-Based CNTs: Exploration for the Carbon Source of CNTs.

Although the preparation of coal-based carbon nanotubes (CNTs) has been realized in many studies, the relationship between carbon source structure of coal and CNT growth has not been studied in depth. In this study, we used lignite and KOH as raw material and catalyst and tuned lignite structure via hydrothermal modification to promote the formation of CNTs during catalytic pyrolysis. The main carbon source of CNTs was explored from the change of coal structure and pyrolysis characteristics. The results indicate that the CNT yield of lignite pyrolysis products is only 2.39%, but the CNT yield increases significantly after lignite was hydrothermally modified in a subcritical water-CO system. The graphitization degree, the order degree, and CNT content increase continuously with the increase in modification temperature, and C-M340 has the highest CNT content of 9.41%. Hydromodification promotes the rearrangement of aromatic carbon structures to generate more condensed aromatic rings linked by short aliphatic chains and aromatic ether bonds. The variation of these structures correlates well with the formation of CNTs and leads to the change in the carbon source components released during coal pyrolysis. Compared to lignite, modified coal releases more aromatic compounds, especially polycyclic aromatic hydrocarbons with ≥3 rings and phenols during catalytic pyrolysis, which is conducive to the transformation into carbon clusters and provides carbon sources for CNT growth. In addition, modified coal releases a slightly more carbon-containing gas (CH4 and CO) than lignite, which has a limited effect on the growth of CNTs. This study provides a novel and efficient method for enhancing the growth of CNTs by a molecular tailoring strategy of coal.

Genome-Wide Identification of LBD Genes in Foxtail Millet (Setaria italica) and Functional Characterization of SiLBD21.

Plant-specific lateral organ boundaries domain (LBD) proteins play important roles in plant growth and development. Foxtail millet (Setaria italica) is one new C4 model crop. However, the functions of foxtail millet LBD genes are unknown. In this study, a genome-wide identification of foxtail millet LBD genes and a systematical analysis were conducted. A total of 33 SiLBD genes were identified. They are unevenly distributed on nine chromosomes. Among these SiLBD genes, six segmental duplication pairs were detected. The thirty-three encoded SiLBD proteins could be classified into two classes and seven clades. Members in the same clade have similar gene structure and motif composition. Forty-seven kinds of cis-elements were found in the putative promoters, and they are related to development/growth, hormone, and abiotic stress response, respectively. Meanwhile, the expression pattern was investigated. Most SiLBD genes are expressed in different tissues, while several genes are mainly expressed in one or two kinds of tissues. In addition, most SiLBD genes respond to different abiotic stresses. Furthermore, the function of SiLBD21, which is mainly expressed in roots, was characterized by ectopic expression in Arabidopsis and rice. Compared to controls, transgenic plants generated shorter primary roots and more lateral roots, indicating the function of SiLBD21 in root development. Overall, our study laid the foundation for further functional elucidation of SiLBD genes.

Identification of Pyrabactin resistance 1-like (PYL) genes in Brachypodium distachyon and functional characterization of BdPYL5.

Abscisic acid (ABA) is an endogenous phytohormone that plays an important role in regulating plant growth, development, and stress response. Pyrabactin resistance 1-like (PYR/PYL) proteins are ABA receptors and core components of ABA signalling in plants. This study identified nine PYL genes in the Brachypodium distachyon genome and they distribute on three chromosomes. Phylogenetical BdPYLs were classified into three clades. 81 protein-protein interactions between 9 BdPYLs and 9 BdPP2C proteins were predicted and 66 pairs were verified by yeast two-hybrid assay previously. Relatively, BdPYL genes are expressed in leaves at high level, and ABA and drought regulate their expression. A homologue of Arabidopsis PYL9, BdPYL5 was selected to overexpress in Arabidopsis to characterize its function. In general, overexpression of BdPYL5 enhanced ABA sensitivity and drought tolerance, implying its conserved function. Our study lays the foundation for further functional elucidation of BdPYL genes.

Flexible biochemical sensors for point-of-care management of diseases: a review.

Health problems have been widely concerned by all mankind. Real-time monitoring of disease-related biomarkers can feedback the physiological status of human body in time, which is very helpful to the diseases management of healthcare. However, conventional non-flexible/rigid biochemical sensors possess low fit and comfort with the human body, hence hindering the accurate and comfortable long-time health monitoring. Flexible and stretchable materials make it possible for sensors to be continuously attached to the human body with good fit, and more precise and higher quality results can be obtained. Thus, tremendous attention has been paid to flexible biochemical sensors in point-of-care (POC) for real-time monitoring the entire disease process. Here, recent progress on flexible biochemical sensors for management of various diseases, focusing on chronic and communicable diseases, is reviewed, and the detection principle and performance of these flexible biochemical sensors are discussed. Finally, some directions and challenges are proposed for further development of flexible biochemical sensors.

Eggshell membrane-mimicking multifunctional nanofiber for in-situ skin wound healing.

Eggshell membrane is a naturally-occurring protective barrier layer for chickens' incubation and shows the close similarity with extracellular matrix. To fully explore and utilize its' structure and active components via a mimicking way will be of great interest for wounds healing. Herein, the well-dispersed CuS nanoparticles were prepared by using eggshell membranes as templates with strong near-infrared absorption and photothermal properties. Furthermore, the as-prepared solution was combined with polyvinyl pyrrolidone and chitosan-derived fluorescent carbon dots for the mimetic synthesis of multifunctional nanofibrous membrane by a hand-held electrospinning device, which has the merits of in-situ operation, the extracellular matrix (ECM)-like architecture, hemostatic, radical scavenging, antibacterial, as well as accelerated healing of skin injury, etc. The electrospun-nanofiber membrane with optimal addition of 100 mg/L CuS nanoparticles was confirmed to be noncytotoxic on human fibroblasts and showed strong antibacterial activities against S. aureus and E. coli under NIR irradiation (980 nm). In addition, the radical scavenging ability was also proved by DPPH experiments. The animal experiments revealed that the nanofiber membrane could accelerate the wound healing process. The work lays down a simple and environmentally-friendly approach for the fabrication and development of promising wound healing materials in skin tissue engineering applications.

Substituent-Induced Electron-Transfer Strategy for Selective Adsorption of N2 in MIL-101(Cr)-X Metal-Organic Frameworks.

N2 removal is of great significance in high-purity O2 production and natural gas purification. Here, we present a substituent-induced electron-transfer strategy for improving N2 capture performance by controlling the Lewis acidity of Cr(III) metal unsaturated sites in Cr-based metal-organic frameworks. With the enhancement of the electron-withdrawing ability of the modified group on terephthalic acid (-NO2 > -CH3), the N2 adsorption ability of MIL-101(Cr)-X was improved significantly. For MIL-101(Cr)-NO2, the adsorption enthalpy of N2 at zero coverage was 30.01 kJ/mol, which was much larger than that of MIL-101(Cr)-CH3 (14.31 kJ/mol). In situ infrared spectroscopy studies, Bader charges, and density functional theory calculations showed that the presence of -NO2 could enhance the Lewis acidity of Cr(III) metal unsaturated sites, which resulted in a strong interaction affinity for N2. The adsorption isotherms indicated that MIL-101(Cr)-NO2 had an excellent N2/O2 (79/21, v/v) selectivity of up to 10.8 and a good N2/CH4 separation performance (SN2/CH4 = 2.8, 298 K, 1 bar). Breakthrough curves showed that MIL-101(Cr)-NO2 had great potential for the efficient separation of N2/O2 and N2/CH4.

Heterogeneous activation of persulfate by Mg doped Ni(OH)2 for efficient degradation of phenol.

Mg doped Ni(OH)2 was synthesized and investigated as an efficient material to activate persulfate (PS) for phenol degradation. The property of the Ni(OH)2 material was enhanced by Mg doping as the removal efficiency of phenol was increased from 74.82 % in Ni(OH)2/PS system to 89.53 % in Mg-doped Ni(OH)2/PS system within 20 min. Such a high removal efficiency revealed that doping Mg into Ni(OH)2 brings about more defects (oxygen vacancies), which facilitated the formation of more active species in the degradation process. The removal efficiencies of phenol increased with the increase of the initial pH from 3 to 11. The influences of Cl-, NO3- and HCO3- on the stability of the system were also studied and the results showed that removal rates of all systems in the presence of these different inorganic anions could reached about 90 % within 20 min. Based on the electron spin resonance (ESR) experiments, 1O2, O2·-, ·OH and SO4•- were identified as the active species in Mg-doped Ni(OH)2/PS system for phenol degradation and a degradation mechanism was proposed for this system. In addition, the as-prepared material retained its activation performance even after 3 repeated cycles.

Efficient Treatment of Phenol Wastewater by Catalytic Ozonation over Micron-Sized Hollow MgO Rods.

Phenol is a nocuous water pollutant that threatens human health and the ecological environment. CoOx-doped micron-sized hollow MgO rods were prepared for the treatment of phenol wastewater by catalytic ozonation. Magnesium sources, precipitants, initial precursor concentration, Co/Mg molar ratio, and catalyst calcination temperature were optimized to obtain the best catalysts. Prepared catalysts were also well characterized by various methods to analyze their structure and physical and chemical properties. In this process, CoOx/MgO with the largest large surface area (151.3 m3/g) showed the best catalytic performance (100 and 79.8% of phenol and chemical oxygen demand (COD) removal ratio, respectively). The hydrolysis of CoOx/MgO plays a positive role in the degradation of phenol. The catalytic mechanism of the degradation of O3 to free radicals over catalysts has been investigated by in situ electronic paramagnetic resonance (EPR). The catalyst can be reused at least five times without any activity decline. The prepared CoOx/MgO catalyst also showed excellent catalytic performance for removal and degradation of ciprofloxacin, norfloxacin, and salicylic acid.

Genistein inhibits the proliferation, migration and invasion of the squamous cell carcinoma cells via inhibition of MEK/ERK and JNK signalling pathways.

PURPOSE: The main purpose of the current research work was to investigate the anticancer activity of Genistein - a plant derived isoflavone - in squamous cell carcinoma SK-MEL-28 (SCC) cells along with studying its effects on cellular apoptosis, DNA damage, cell migration and invasion and MEK/ERK/JNK signalling pathway. METHODS: Cell proliferation was examined by CCK-8 (Cell Counting Kit-8) assay while the effects on apoptosis were evaluated by DAPI staining and Comet assay using fluorescence microscopy. Transwell assay was used for checking the effects on cell migration and invasion while western blot method was used to evaluate the effects on the expression of MEK/ERK/JNK proteins. RESULTS: The results showed that Genistein led to dose-dependent cytotoxic effects in these cells showing an IC50 value of 14.5 µM. It also led to dose-dependent apoptosis and induced DNA damage as shown by fluorescence microscopy. Genistein also inhibited cell migration and invasion dose-dependently, along with inhibiting matrix metalloproteinase (MMP)-9 expression. Genistein also led to inhibition of the expression of p-JNK with no apparent effects on the total JNK expression. It also showed significant and dose-dependent inhibition of the expression of p-MEK and p-ERK proteins. CONCLUSIONS: Genistein has a significant anticancer activity in SK-MEL-28 human SCC cells, inducing apoptosis, DNA damage, cell migration and invasion and inhibiting MEK/ERK and JNK signalling pathway.

The tissue distribution and excretion study of mosapride and its active des-p-fluorobenzyl and 4'-N-oxide metabolites in rats by ultra-high performance liquid chromatography-tandem mass spectrometry method.

1. Mosapride is a potent gastroprokinetic agent, and des-p-fluorobenzyl mosapride (M1) and mosapride-N-oxide (M2) are its two major active metabolites.2. The validated ultra-high performance liquid chromatography-tandem mass spectrometry method was successfully applied to the distribution and excretion of mosapride and its two active metabolites.3. Mosapride and its metabolites were distributed widely and rapidly in various tissues. The highest concentration of mosapride and M2 in both male and female rats was found in the duodenum, followed by cecum.4. The excretion study showed that a total of 71.8% (37.6, 22.4 and 11.8% for urine, feces and bile, respectively) and 66.3% (35.7, 22.8 and 7.8% for urine, feces and bile) of administered dose was recovered from male and female excreta. M1 was excreted in the largest dose percentage, followed by mosapride and M2, and the total cumulative excretion amounts were about 36.9, 28.1 and 11.6% in male rat, while 24.3, 25.9 and 16.2% in female rat. The results demonstrated for the first time that M2 is one of the important excretion forms of mosapride, which is much higher than that of mosapride in urine.5. This work could provide valuable information for further pharmacological and clinical studies of mosapride.

An available strategy based on accurate mass by ultra high performance liquid chromatography coupled to Fourier transform ion cyclotron resonance mass spectrometry technology to characterization of metabolic profile of palbociclib in rat urine, feces and bile.

Palbociclib (named PD 0332991) is a novel highly selective cyclin-dependent kinase 4 and 6 (CDK 4/6) inhibitor, which has been approved by the Food and Drug Administration (FDA) for the treatment of hormone-receptor-positive advanced breast cancer. This present study developed a comprehensive strategy to investigate the metabolic profile of palbociclib in rat urine, feces and bile samples based on an ultra high performance liquid chromatography coupled to Fourier transform ion cyclotron resonance mass spectrometry (UHPLC-FT-ICR MS). A total of 29 metabolites, including 18 phase I metabolites and 11 phase II metabolites, were detected and identified. The metabolic pathways included hydroxylation, oxidation, dehydrogenation, N-dealkylation, carbonylation, oxidative deamination, acetylation, glucuronidation, sulphate conjugation as well as the crossover of multiple metabolic pathways in vivo, and 16 of these metabolites were proposed for the first time. This study showed an insight into the metabolism of palbociclib in vivo, which may provide relevant chemical information for subsequent studies in the future.

The synthesis and biological evaluation of a new bioactive metabolite of mosapride as a potential gastroprokinetic agent.

AIM: To synthesize the new bioactive metabolites of mosapride (R)-N-[2-hydroxy-3-(4-fluorobenzyl)amino]-propyl-5-chlorine-4-amino-2-ethoxyben-zamide (R-isomer) and (S)-N-[2-hydroxy-3-(4-fluorobenzyl)amino]-propyl-5-chlorine-4-amino-2-ethoxybenzamide (S-isomer) and evaluate their in vitro and in vivo pharmacological and pharmacokinetic profiles. RESULTS: S-isomer as a gastroprokinetic agent showed significant pharmacological activities in vivo. Furthermore, compared with the EC50 values for R-isomer and mosapride, S-isomer was proven to generate the same 5-HT4 receptor agonistic activity with a smaller amount. S-isomer exhibited significant differences in the pharmacokinetic properties, which indicate that higher absorption rate and extent compared with R-isomer. CONCLUSION: S-isomer might have great potential as a safe and effective prokinetic agent capable of lessening gastrointestinal symptoms and increasing quality of life.