Hybrid Vibration and UV Fluorescence Technology for Rapid Imaging and Guidance for Manual Removal of Fish Bones from Fish Floss.

The objective of the proposed human-machine cooperation (HMC) workstation is to both rapidly detect calcium-based fish bones in masses of minced fish floss and visually guide operators in approaching and removing the detected fish bones by hand based on the detection of fingernails or plastic-based gloves. Because vibration is a separation mechanism that can prevent absorption or scattering in thick fish floss for UV fluorescence detection, the design of the HMC workstation included a vibration unit together with an optical box and display screens. The system was tested with commonly used fish (swordfish, salmon, tuna, and cod) representing various cooking conditions (raw meat, steam-cooked meat, and fish floss), their bones, and contaminating materials such as derived from gloves made of various types of plastic (polyvinylchloride, emulsion, and rubber) commonly used in the removal of fish bones. These aspects were each investigated using the spectrum analyzer and the optical box to obtain and analyze the fluorescence spectra and images. The filter was mounted on a charge-coupled device, and its transmission-wavelength window was based on the characteristic band for fish bones observed in the spectra. Gray-level AI algorithm was utilized to generate white marker rectangles. The vibration unit supports two mechanisms of air and downstream separation to improve the imaging screening of fish bones inside the considerable flow of fish floss. Notably, under 310 nm ultraviolet B (UVB) excitation, the fluorescence peaks of the raw fillets, steam-cooked meat, and fish floss were observed at for bands at longer wavelengths (500-600 nm), whereas those of the calcium and plastic materials occurred in shorter wavelength bands (400-500 nm). Perfect accuracy of 100% was achieved with the detection of 20 fish bones in 2 kg of fish floss, and the long test time of around 10-12 min results from the manual removal of these fish bones.

Analysis of heterocyclic amines in meat products by liquid chromatography - Tandem mass spectrometry.

Heterocyclic amines (HCAs), a class made up of more than 25 compounds, are unintended hazardous substances that are generated by the heating or processing of proteinaceous foods at high temperatures. The International Agency for Research on Cancer (IARC) has classified four such HCAs (IQ, MeIQ, MeIQx, and PhIP) as being probable or possible human carcinogens. In this study, two sample preparation strategies, liquid-liquid extraction (LLE) with solid-phase extraction (SPE) and a rapid, easy, cheap, effective, rugged, and safe extraction (QuEChERS) method, were investigated for the determination of 11 types of HCAs in meat products by LC-MS/MS. The HCAs in the samples were first extracted with acetonitrile by LLE, and followed by SPE. In the case of QuEChERS extraction, acetonitrile is used as the LLME solvent, and PSA, C18EC and MgSO4 were used as the dSPE sorbent. Both methods showed good performance with respect to precision (RSD < 15.15%), accuracy (79.80-117.64%), recovery (52.39-116.88%), limit of quantitation for a spiked meat extract (0.01-10 ppb) and correlation coefficients (>0.993). The QuEChERS extraction strategy provided a better linear dynamic range and superior sensitivity in comparison with the LLE-SPE approach. HCAs were successfully quantified in real samples using the two proposed approaches by LC-MS/MS.

Concentration variation and molecular characteristics of soluble (1,3;1,6)-ß-D-glucans in submerged cultivation products of Ganoderma lucidum mycelium.

(1,3)-ß-D-Glucans with (1,6)-ß-D-glucosyl branches are bioactive polysaccharides in fruiting bodies and mycelia of Ganoderma lucidum, a mushroom used in traditional Chinese medicine. Submerged cultivation of mycelium is one of the more efficient means of generating polysaccharides from this fungus. Twelve mycelium samples examined in this study demonstrated the quantitative and qualitative molecular characteristics of soluble (1,3;1,6)-ß-D-glucans. It was observed that the concentration of soluble (1,3;1,6)-ß-D-glucan varied substantially from 1.3 to 79.9 mg/dL. (1,3;1,6)-ß-D-Glucans also preserved their molecular characteristics with degrees of branching (DB) of 0.21-0.36 and molecular masses of 10(5)-10(6) g/mol for those samples with substantial quantities of ß-D-glucan. Using the high aggregating tendency of these molecules, (1,3;1,6)-ß-D-glucans were successfully purified via fractional precipitation with 35% (v/v) ethanol. (1,3;1,6)-ß-D-Glucan was proposed as a putative bioactive marker for immunomodulation because it was the most abundant polysaccharide in G. lucidum mycelium products to stimulate macrophage RAW 264.7 cells to release TNF-α.

Establishment of reporter platforms capable of detecting NF-κB mediated immuno-modulatory activity.

Cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) are transcriptional targets of nuclear factor kappa B (NF-κB) that are involved in inflammatory responses. The aim of this study is to develop a method for efficiently detecting inflammation modulatory activities. Here we established RAW264.7 macrophage cells stably expressing a luciferase reporter gene directed by iNOS or COX-2 promoter. Lipopolysaccharide (LPS) treatment stimulated the luciferase activity which paralleled with increased iNOS and COX-2 mRNA levels determined by RT-q-PCR. The LPS-stimulated luciferase activity was blocked by NF-κB inhibitor CAPE and by nobiletin, an anti-inflammatory natural product from citrus peels. We have applied the platforms to screen various mushroom species; analysis by scatter plot revealed a strong correlation to the results obtained by ELISA-based detection of TNF-α. Together we have established luciferase reporter systems sensitive to NF-κB-dependent iNOS and COX-2 activation, which provides an alternative screening method for identifying food components with immune-modulatory activities.