Survival of MNV-1, <i>E. coli</i> K-12 and <i>L. innocua</i> on different contaminated tableware before and after 1 hour air-drying at 24±2°C.

<p>Survival of MNV-1, <i>E. coli</i> K-12 and <i>L. innocua</i> on different contaminated tableware before and after 1 hour air-drying at 24±2°C.</p

Inactivation of MNV-1 in stock solution and in inoculated milk by the Control, Chlorine (200 ppm) and QAC sanitizing (200 ppm) solutions at 49°C for 10 sec.

<p>Inactivation of MNV-1 in stock solution and in inoculated milk by the Control, Chlorine (200 ppm) and QAC sanitizing (200 ppm) solutions at 49°C for 10 sec.</p

Devices used during manual ware-washing to clean the different tableware items.

<p>a) Cylindrical sponge b) Sponge attached to the spring-loaded tool.</p

Survival of MNV-1, <i>E. coli</i> K-12 and <i>L. innocua</i> on contaminated tableware items after washing and sanitizing, using the mechanical dishwasher.

<p>Survival of MNV-1, <i>E. coli</i> K-12 and <i>L. innocua</i> on contaminated tableware items after washing and sanitizing, using the mechanical dishwasher.</p

Survival of MNV-1, <i>E. coli</i> K-12 and <i>L. innocua</i> on contaminated tableware items after washing and sanitizing, during manual ware-washing.

<p>Survival of MNV-1, <i>E. coli</i> K-12 and <i>L. innocua</i> on contaminated tableware items after washing and sanitizing, during manual ware-washing.</p

Label-Free On-Chip Selective Extraction of Cell-Aggregate-Laden Microcapsules from Oil into Aqueous Solution with Optical Sensor and Dielectrophoresis

Microfluidic encapsulation of cells or tissues in biocompatible solidlike hydrogels has wide biomedical applications. However, the microfluidically encapsulated cells/tissues are usually suspended in oil and need to be extracted into aqueous solution for further culture or use. Current extracting techniques are either nonselective for the cell/tissue-laden hydrogel microcapsules or rely on fluorescence labeling of the cells/tissues, which may be undesired for their further culture or use. Here we developed a microelectromechanical system (MEMS) to achieve label-free on-chip selective extraction of cell-aggregate-laden hydrogel microcapsules from oil into aqueous solution. The system includes a microfluidic device, an optical sensor, a dielectrophoretic (DEP) actuator, and microcontrollers. The microfluidic device is for encapsulating cell aggregates in hydrogel microcapsules using the flow-focusing function with microchannels for extracting microcapsules. The optical sensor is to detect the cell aggregates, based on the difference of the optical properties between the cell aggregates and surrounding solution before their encapsulation in hydrogel microcapsules. This strategy is used because the difference in optical property between the cell-aggregate-laden hydrogel microcapsules and empty microcapsules is too small to tell them apart with a commonly used optical sensor. The DEP actuator, which is controlled by the sensor and microcontrollers, is for selectively extracting the targeted hydrogel microcapsules by DEP force. The results indicate this system can achieve selective extraction of cell-aggregate-laden hydrogel microcapsules with ∼100% efficiency without compromising the cell viability, and can improve the purity of the cell-aggregate-laden microcapsules by more than 75 times compared with nonselective extraction

Simultaneous Multiplexed Stripping Voltammetric Monitoring of Marine Toxins in Seafood Based on Distinguishable Metal Nanocluster-Labeled Molecular Tags

Marine toxins from microscopic algae can accumulate through the food chain and cause various neurological and gastrointestinal illnesses for human health. Herein, we designed a new ultrasensitive multiplexed immunoassay protocol for simultaneous electrochemical determination of brevetoxin B (BTX-2) and dinophysistoxin-1 (DTX-1) in seafood using distinguishable metal nanocluster-labeled molecular tags as traces on bifunctionalized magnetic capture probes. To construct such a bifunctionalized probe, monoclonal mouse anti-BTX-2 (mAb₁) and anti-DTX-1 (mAb₂) antibodies were co-immobilized on a magnetic bead (MB–mAb_1,2). The distinguishable metal nanoclusters including cadmium nanoclusters (CdNC) and copper nanoclusters (CuNC) were synthesized using the artificial peptides with amino acid sequence CCCYYY, which were used as distinguishable signal tags for the label of the corresponding bovine serum albumin–BTX-2 and bovine serum albumin–DTX-1 conjugates. A competitive-type immunoassay format was adopted for the online simultaneous monitoring of BTX-2 and DTX-1 on a homemade flow-through magnetic detection cell. The assay was based on the stripping voltammetric behaviors of the labeled CdNC and CuNC at the various peak potentials in pH 2.5 HCl containing 0.01 M KCl using square wave anodic stripping voltammetry (SWASV). Under optimal conditions, the multiplexed immunoassays enabled simultaneous detection of BTX-2 and DTX-1 in a single run with wide working ranges of 0.005–5 ng mL^–1 for two marine toxins. The limit of detection (LOD) and limit of quantification (LOQ) were 1.8 and 6.0 pg mL^–1 for BTX-2, while those for DTX-1 were 2.2 and 7.3 pg mL^–1, respectively. No non-specific adsorption and electrochemical cross-talk between neighboring sites were observed during a series of procedures to detect target analytes. The covalent conjugation of biomolecules onto the nanoclusters and magnetic beads resulted in good repeatability and intermediate precision down to 9.5%. The method featured unbiased identification of negative (blank) and positive samples. No significant differences at the 0.05 significance level were encountered in the analysis of 12 spiked samples, including Sinonovacula constricta, Musculista senhousia, and Tegillarca granosa, between the multiplexed immunoassay and commercially available enzyme-linked immunosorbent assay (ELISA) for analysis of BTX-2 and DTX-1

Label-Free On-Chip Selective Extraction of Cell-Aggregate-Laden Microcapsules from Oil into Aqueous Solution with Optical Sensor and Dielectrophoresis

Microfluidic encapsulation of cells or tissues in biocompatible solidlike hydrogels has wide biomedical applications. However, the microfluidically encapsulated cells/tissues are usually suspended in oil and need to be extracted into aqueous solution for further culture or use. Current extracting techniques are either nonselective for the cell/tissue-laden hydrogel microcapsules or rely on fluorescence labeling of the cells/tissues, which may be undesired for their further culture or use. Here we developed a microelectromechanical system (MEMS) to achieve label-free on-chip selective extraction of cell-aggregate-laden hydrogel microcapsules from oil into aqueous solution. The system includes a microfluidic device, an optical sensor, a dielectrophoretic (DEP) actuator, and microcontrollers. The microfluidic device is for encapsulating cell aggregates in hydrogel microcapsules using the flow-focusing function with microchannels for extracting microcapsules. The optical sensor is to detect the cell aggregates, based on the difference of the optical properties between the cell aggregates and surrounding solution before their encapsulation in hydrogel microcapsules. This strategy is used because the difference in optical property between the cell-aggregate-laden hydrogel microcapsules and empty microcapsules is too small to tell them apart with a commonly used optical sensor. The DEP actuator, which is controlled by the sensor and microcontrollers, is for selectively extracting the targeted hydrogel microcapsules by DEP force. The results indicate this system can achieve selective extraction of cell-aggregate-laden hydrogel microcapsules with ∼100% efficiency without compromising the cell viability, and can improve the purity of the cell-aggregate-laden microcapsules by more than 75 times compared with nonselective extraction

Complex Disease Networks of Trait-­‐Associated SNPs Unveiled by Information Theory

Objective: Thousands of complex disease SNPs have been discovered in Genome Wide Association Studies (GWAS). However, these intragenic SNPs have not been collectively mined to unveil the genetic architecture between complex clinical traits. We hypothesize that biological annotations of host genes of trait-associated SNPs may reveal the biomolecular modularity across complex disease traits and offer insights for drug repositioning. Methods: In this study, we used trait-to-polymorphism (SNPs) associations confirmed in GWAS. We developed a novel method to quantify trait-trait similarity anchored in Gene Ontology annotations of human proteins and information theory. We then validated these results with the shortest paths of physical protein interactions between biologically similar traits. Results: We constructed a network consisting of 280 significant intertrait similarities among 177 disease traits, which covered 1,438 well-validated disease-associated SNPs. 39% of intertrait connections were confirmed by curators and the following additional studies demonstrated the validity of a proportion of the remainder. On a phenotypic trait level, higher Gene Ontology similarity between proteins correlated with smaller "shortest distance" in protein interaction networks of complexly inherited diseases (Spearman p<2.2x10-16). Further, "cancer traits" were similar to one another, as were "metabolic syndrome traits"(FET p=0.001 and 3.5x10-7). Conclusion: We report an imputed disease network by information-anchored functional similarity from GWAS trait-associated SNPs. We also demonstrate that small shortest path of protein interactions correlates with complex disease function. Taken together, these findings provide the framework for investigating drug targets with unbiased functional biomolecular networks rather than worn-out single gene and subjective canonical pathway approaches

Fish Oil Ameliorates Vibrio parahaemolyticus Infection in Mice by Restoring Colonic Microbiota, Metabolic Profiles, and Immune Homeostasis

The effect of fish oil (FO) on colonic function, immunity, and microbiota was investigated in Vibrio parahaemolyticus (Vp)-infected C57BL/6J mice. Mice intragastrically presupplemented with FO (4.0 mg) significantly reduced Vp infection as evidenced by stabilizing body weight and reducing disease activity index score and immune organ ratios. FO minimized colonic pathological damage, strengthened the mucosal barrier, and sustained epithelial permeability by increasing epithelial crypt depth, goblet cell numbers, and tight junctions and inhibiting colonic collagen accumulation and fibrosis protein expression. Mechanistically, FO enhanced immunity by decreasing colonic CD3+ T cells, increasing CD4+ T cells, downregulating the TLR4 pathway, reducing interleukin-17 (IL-17) and tumor necrosis factor-α, and increasing immune cytokine IL-4 and interferon-γ levels. Additionally, FO maintained colonic microbiota eubiosis by improving microbial diversity and boosting Clostridium, Akkermansia, and Roseburia growth and their derived propionic acid and butyric acid levels. Collectively, FO alleviated Vp infection by enriching beneficial colonic microbiota and metabolites and restoring immune homeostasis