Strain-specific Recovery of S. sonnei from Artificially Contaminated Baby Carrots: Enhancing Food-safety Investigations with a Customized Shigella Detection Method Based on Genomically predicted Antibiotic Resistance Traits.

Shigella spp. are Gram-negative gastrointestinal bacterial pathogens that cause bacillary dysentery or shigellosis in humans. Isolation of Shigella from outbreak-associated foods is often problematic due to the lack of selectivity of cultural enrichment broths. To facilitate Shigella recovery from foods, we have developed strain-specific enrichment media based on the genomically-predicted antimicrobial resistance (AMR) features of an outbreak-associated Shigella sonnei strain harboring resistance genes for streptomycin (STR) and trimethoprim (TMP). To assess performance of the method, baby carrots were artificially contaminated with the S. sonnei strain at low (2.4 CFU), medium (23.5 CFU), and high levels (235 CFU) along with 10-fold higher levels of a Shigella-inhibiting Escherichia coli strain. The target S. sonnei strain was successfully recovered from artificially-contaminated baby carrots when enriched in modified Tryptone Soya Broth (mTSB) supplemented with TMP, whereas Shigella was not recovered from Shigella broth (SB) or SB supplemented with STR. Quantitative PCR analysis indicated that supplementation of the enrichment broths with TMP or STR increased the relative proportion of S. sonnei in enrichment cultures, except at the lowest inoculation level for STR. Microbiome profiling of the baby carrot enrichment cultures conducted by 16S rRNA gene sequencing indicated that both SB-STR and mTSB-TMP repressed the growth of competing Enterobacteriaceae in the enrichment cultures, relative to SB without supplementation. Overall, improved Shigella recovery was achieved with the addition of the appropriate custom selective agent during cultural enrichments demonstrating that genomically informed custom selective enrichment of Shigella could be a valuable tool for supporting future foodborne shigellosis outbreak investigations.

Overcoming Microbial Inhibition of S. Sonnei Through the Exploitation of Genomically Predicted Antibiotic Resistance Profiles for the Development of Food Enrichment Media.

Linking outbreaks of Shigella spp. to specific foods is challenging due to poor selectivity of current enrichment media. We have previously shown that enrichment media, tailored to the genomically-predicted antimicrobial resistance (AMR) of Shiga toxigenic E. coli strains, enhances their isolation from foods. This study investigates the application of this approach for Shigella isolation. The AMR gene profiles of 21,908 published S. sonnei genomes indicated a high prevalence of genes conferring resistance to streptomycin (aadA, aph(3″)-Ib, aph(6)-Id, 92.8%), sulfonamides (sul1, sul2, 74.8%), and/or trimethoprim (dfrA, 96.2%). Genomic analysis and antibiotic susceptibility testing conducted with a panel of 17 outbreak-associated S. sonnei strains confirmed the correlation of AMR gene detection with resistance phenotypes. Supplementation of Shigella Broth (SB) with up to 400 µg/mL of trimethoprim or sulfadiazine did not suppress the growth of sensitive strains, whereas 100 µg/mL of streptomycin increased the selectivity of this broth. All three antibiotics increased the selectivity of modified Tryptone Soya Broth (mTSB). Based on these results, supplemented media formulations were developed and assessed by measuring the relative growth of S. sonnei in cultures coinoculated with a strain of bacteriocin-producing E. coli that is inhibitory to Shigella growth. S. sonnei was not recovered from cocultures grown in SB or mTSB without antibiotics. In contrast, media supplemented with streptomycin at 50 and 100 µg/mL, trimethoprim at 25 and 50 µg/mL, and sulfadiazine at 100 µg/mL increased the relative proportion of S. sonnei in postenrichment cultures. The enhanced recovery of resistant S. sonnei strains achieved in this study indicates that, in cases where genomic data are available for clinical S. sonnei isolates, customization of selective enrichment media based on AMR gene detection could be a valuable tool for supporting the investigation of foodborne shigellosis outbreaks.

Matrine reduced intestinal stem cell damage in eimeria necatrix-infected chicks via blocking hyperactivation of Wnt signaling.

BACKGROUND: Coccidiosis is a rapidly spreading and acute parasitic disease that seriously threatening the intestinal health of poultry. Matrine from leguminous plants has anthelmintic and anti-inflammatory properties. PURPOSE: This assay was conducted to explore the protective effects of Matrine and the AntiC (a Matrine compound) on Eimeria necatrix (EN)-infected chick small intestines and to provide a nutritional intervention strategy for EN injury. STUDY DESIGN: The in vivo (chick) experiment: A total of 392 one-day-old yellow-feathered broilers were randomly assigned to six groups in a 21-day study: control group, 350 mg/kg Matrine group, 500 mg/kg AntiC group, EN group, and EN + 350 mg/kg Matrine group, EN + 500 mg/kg AntiC group. The in vitro (chick intestinal organoids, IOs): The IOs were treated with PBS, Matrine, AntiC, 3 µM CHIR99021, EN (15,000 EN sporozoites), EN + Matrine, EN + AntiC, EN + Matrine + CHIR99021, EN + AntiC + CHIR99021. METHODS: The structural integrity of chicks jejunal crypt-villus axis was evaluated by hematoxylin and eosin (H&E) staining and transmission electron microscopy (TEM). And the activity of intestinal stem cells (ISCs) located in crypts was assessed by in vitro expansion advantages of a primary in IOs model. Then, the changes of Wnt/ß-catenin signaling in jejunal tissues and IOs were detected by Real-Time qPCR,Western blotting and immunohistochemistry. RESULTS: The results showed that dietary supplementation with Matrine or AntiC rescued the jejunal injury caused by EN, as indicated by increased villus height, reduced crypt hyperplasia, and enhanced expression of tight junction proteins. Moreover, there was less budding efficiency of the IOs expanded from jejunal crypts of chicks in the EN group than that in the Matrine and AntiC group, respectively. Further investigation showed that AntiC and Matrine inhibited EN-stimulated Wnt/ß-catenin signaling. The fact that Wnt/ß-catenin activation via CHIR99021 led to the failure of Matrine and AntiC to rescue damaged ISCs confirmed the dominance of this signaling. CONCLUSION: Our results suggest that Matrine and AntiC inhibit ISC proliferation and promote ISC differentiation into absorptive cells by preventing the hyperactivation of Wnt/ß-catenin signaling, thereby standardizing the function of ISC proliferation and differentiation, which provides new insights into mitigating EN injury by Matrine and AntiC.

J-Aggregation induced NIR-II fluorescence: an aza-BODIPY luminogen for efficient phototheranostics.

The development of organic dyes with emission peaks in the second near-infrared window (NIR-II 1000-1700 nm) is highly desirable for in vivo imaging and imaging-guided phototheranostics. However, the lack of appropriate molecular frameworks and the challenges associated with complex synthesis critically hinder the development of new candidate fluorophores. J-Aggregation is considered as a smart and straightforward way to construct such a therapeutic agent with NIR-II fluorescence imaging properties. Here, we present the design and synthesis of an aza-BODIPY probe (TA). Upon encapsulation within the amphiphilic polymer DSPEG-PEG2000-NH2, TA underwent self-assembly and formed J-aggregates (TAJ NPs), which showed emission at 1020 nm. High spatial resolution and adequate signal-to-noise ratio of the TAJ NPs are demonstrated for noninvasive bioimaging of the vasculature, lymph nodes and bones of mice in the NIR-II region. Moreover, the TAJ NPs exhibited good tumor enrichment efficiency with reduced liver accumulation and significant imaging-guided phototherapy performance against lung cancer cells. Taken together, this work not only introduces a new NIR-II imaging and phototheranostic agent based on J-aggregates, but also provides insight into the development of versatile organic dyes for future clinical implementation.

Exploiting Cancer Vulnerabilities by Blocking of the DHODH and GPX4 Pathways: A Multifunctional Bodipy/PROTAC Nanoplatform for the Efficient Synergistic Ferroptosis Therapy.

Ferroptosis is a form of programmed cell death and plays an important role in many diseases. Dihydroorotate dehydrogenase (DHODH) and glutathione peroxidase 4 (GPX4) play major roles in cell resistance to ferroptosis. Therefore, inactivation of these proteins provides an excellent opportunity for efficient ferroptosis-based synergistic cancer therapy. In this study, a multifunctional nanoagent (BPNpro ) containing a GPX4 targeting boron dipyrromethene (Bodipy) probe (BP) and a DHODH targeting proteolysis targeting chimera (PROTAC) is reported. BPNpro is prepared using a nanoprecipitation method in the presence of a thermoresponsive liposome, where BP is encapsulated inside and the cathepsin B (CatB)-cleavable PROTAC peptide (DPCP) is modified on the outer surface. In the presence of near-infrared (NIR) photoirradiation, BPNpro is melted and BP is released in tumor cells. Subsequently, BP inhibits the activity of GPX4 by covalently bonding with the selenocysteine at the enzyme active site. In addition, DPCP achieves sustained degradation of DHODH upon activation by CatB overexpressed in the tumor. The synergistic deactivation of GPX4 and DHODH induces extensive ferroptosis and subsequent cell death. In vivo and in vitro studies clearly show that the proposed ferroptosis therapy provides excellent antitumor effect.

Constructing mesoporous Zr-doped SiO2 onto efficient Z-scheme TiO2/g-C3N4 heterojunction for antibiotic degradation via adsorption-photocatalysis and mechanism insight.

Novel modified-TiO2/Zr-doped SiO2/g-C3N4 ternary composite is fabricated via an in-situ grow of porous Zr-SiO2 layer to TiO2/g-C3N4 heterojunction, which exhibits well adsorption-photocatalytic performance under simulated solar light irradiation. The nano-size mesoporous TiO2 are dispersed on the lamellar g-C3N4, and the Zr-SiO2 is in-situ fabricated onto the surface of g-C3N4 sheets. The adsorption occurs on the SiO2 layers, and doping Zr element to SiO2 enhances the adsorption of pollutants, while the photocatalytic reaction occurs on the valence band (VB) of TiO2 and conduction band (CB) of g-C3N4, which gives reactive oxygen species of ∙O2-, h+, and ∙OH for high efficient decomposition of antibiotics, i.e. berberine hydrochloride (98.11%), tetracycline (80.76%), and oxytetracycline (84.84%). The excellent adsorption capacity and Z-scheme photoinduced charge carrier migration behavior endowed the novel material with enhanced berberine hydrochloride (BH) removal in water, which approximately 2.5 and 3.8 folds than that of pure g-C3N4 and sole TiO2, respectively. Three degradation pathways are unraveled by LC-MS and theoretical calculations. Furthermore, the toxicity of intermediates was evaluated by the Toxicity Estimation Software Tool (T.E.S.T.), the result demonstrated a good application potential of M-TiO2/Zr-SiO2/g-C3N4 as an novel adsorptive photocatalyst.

Anatase quantum dots decorated silica/carbon lamellas for removal of antipsychotic drugs via adsorption-photocatalysis and toxicity evaluation.

In this work, discrete quantum dots of crystallized anatase TiO2 are successfully anchored on carbon nanosheets containing amorphous SiO2 via templated self-assembly and pyrolysis routes. The novel hybrid photocatalyst of TiO2/C/SiO2 exhibits well coupled adsorption and visible light photocatalysis on chlorpromazine (CPZ) and the rate constants are 0.0223 and 0.0198 min-1, respectively. The direct photocatalytic degradation of CPZ under static conditions reaches 91.1% within 3 h while a removal rate of 31.4% for CPZ could be retained under dynamic flow conditions, and the improved performance could be attributed to enhanced adsorption via SiO2/C and highly exposure of TiO2 QDs surface. Based on the trapping experiments, ESR, LC-MS, and toxicity evaluation, O2- free radicals are identified as main reactive species for CPZ degradation along three possible pathways, with reduced toxicities for its intermediates. The cell viability tests of photocatalytic-degraded solutions and the catalyst exhibit negligible toxicities for both intermediates and the material, suggesting the novel composite of TiO2/C/SiO2 as an environmental friendly photocatalyst for pharmaceutical wastewater treatment.

Effect of the homogenization technique on the enumeration of psychrotrophic bacteria in food absorbent pads.

Four methods were tested for enumerating bacteria present in the absorbent pads (AP) used in packaging chicken and other meats. Viable counts were ascertained at day 0 and day 7 (d0 and d7, respectively). Sampling bacterial cells from AP were carried out using a countertop blender, Stomacher, sonication, and blender in combination to sonication. The release of bacterial cells by breaking down the AP with the blender resulted in the highest CFU counts. At d0, a bacterial recovery rate of 94% was obtained with the blender, while the recovery rates using Stomacher or sonication alone were 58% and 73%, respectively. At d7, the Stomacher treatment also gave the lowest colony forming unit (CFU) values in the AP incubated at 7 °C. Sonication of the AP prior to homogenization with the blender did not increase CFU counts. Results suggested that breaking down the AP with a blender gives higher CFU levels than the Stomacher, which is the most commonly used technique for this purpose.

CuCo2O4/N-Doped CNTs loaded with molecularly imprinted polymer for electrochemical sensor: Preparation, characterization and detection of metronidazole.

CuCo2O4 nanoparticles modified with nitrogen doped carbon nanotubes (CuCo2O4/N-CNTs) have high specific surface area and good electrical conductivity. Herein, a novel electrochemical sensor based on CuCo2O4/N-CNTs loaded molecularly imprinted polymer (MIP) modified glassy carbon electrode (GCE) is proposed for rapid and ultrasensitive detection of metronidazole (MNZ). The composite of CuCo2O4/N-CNTs with MIP significantly enhances the electrical signal. The electrochemical polymerization was performed with MNZ as template and aniline as functional monomer by cyclic voltammetry (CV), and differential pulse voltammetry (DPV) was used to detect MNZ. Factors that affect sensor response were optimized. Under the optimal experimental conditions, the DPV current response shows two linearity ranges for MNZ in the range of 0.005-0.1 µM and 0.1-100 µM with very low limit of detection (LOD) of 0.48 nM (S/N = 3). This electrochemical sensing system has high sensitivity, selectivity, excellent reproducibility, repeatability and stability. The recovery (95.9%-100.9%) and reasonable relative standard deviation (RSD) (3.2%-4.8%) for determination of real samples indicate the practicality of the sensing system. This sensing system has high potential for rapid determination of MNZ in samples such as metronidazole tablets, human serum and urine.

Optical Methods in Fingerprint Imaging for Medical and Personality Applications.

Over the years, analysis and induction of personality traits has been a topic for individual subjective conjecture or speculation, rather than a focus of inductive scientific analysis. This study proposes a novel framework for analysis and induction of personality traits. First, 14 personality constructs based on the "Big Five" personality factors were developed. Next, a new fingerprint image algorithm was used for classification, and the fingerprints were classified into eight types. The relationship between personality traits and fingerprint type was derived from the results of the questionnaire survey. After comparison of pre-test and post-test results, this study determined the induction ability of personality traits from fingerprint type. Experimental results showed that the left/right thumbprint type of a majority of subjects was left loop/right loop and that the personalities of individuals with this fingerprint type were moderate with no significant differences in the 14 personality constructs.

[Construction and expression of an eukaryotic recombinant plasmid from a hybrid gene of antigen for Plasmodium falciparum].

OBJECTIVE: To construct an eukaryotic recombinant plasmid with HGFSP, a hybrid gene encoding the antigen epitopes of MSA1, MSA2, RESA and CSP in different developing stages of Plasmodium falciparum (P. f.). METHODS: HGFSP was sub-cloned into an eukaryotic expression plasmid pcDNA3 from a prokaryotic recombinant plasmid pSK-HGFSP to construct the eukaryotic recombinant expression plasmid pc-HGFSP. The identified recombinant was then transfected into HepG2 cells with liposome-mediated method. The G418-selected positive cell clones were tested to identify the immunogenicity of HGFSP-expressing antigen. RESULTS: It was evidenced that HGFSP was correctly inserted into pcDNA3 by restriction enzymes map analysis. HGFSP-expressing antigen-specific fluorescent response was observed in pc-HGFSP-transfected HepG2 cells. The results of SDS-PAGE and Western-blotting showed that there was a 23 kD protein band, which can be specifically recognized by anti-sera of HGFSP-expressing protein in pc-HGFSP-transfected HepG2 cell lysis. CONCLUSION: Pc-HGFSP, an eukaryotic recombinant plasmid encoding hybrid antigen epitopes of P. f., was constructed successfully and the antigenicity of pc-HGFSP-expressing protein was confirmed.