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The three-dimensional fluorescence spectroscopy features the advantage of obtaining emission spectra at different excitation wavelengths and providing more detailed information. This study established a simple method to discriminate both the producer and grade of matcha tea by coupling three-dimensional fluorescence spectroscopy analysis and distance discrimination. The matcha tea was extracted three times and three-dimensional fluorescence spectroscopies of these tea infusions were scanned; then, the dimension of three-dimensional fluorescence spectroscopies was reduced by the integration at three specific areas showing local peaks of fluorescence intensity, and a series of vectors were constructed based on a combination of integrated vectors of the three tea infusions; finally, four distances were used to discriminate the producer and grade of matcha tea, and two discriminative patterns were compared. The results indicated that proper vector construction, appropriate discriminative distance, and correct steps are three key factors to ensure the high accuracy of the discrimination. The vector based on the three-dimensional fluorescence spectroscopy of all three tea infusions resulted in a higher accuracy than those only based on spectroscopy of one or two tea infusions, and the first tea infusion was more sensitive than the other tea infusion. The Mahalanobis distance had a higher accuracy that was up to 100% when the vector is appropriate, while the other three distances were about 60-90%. The two-step discriminative pattern, identifying the producer first and the grade second, showed a higher accuracy and a smaller uncertainty than the one-step pattern of identifying both directly. These key conclusions above help discriminate the producer and grade of matcha in a quick, accurate, and green method through three-dimensional fluorescence spectroscopy, as well as in quality inspections and identifying the critical parameters of the producing process.
Introduction Cochlear spiral ganglion neurons (SGNs) could be damaged by ototoxic drug, cisplatin (Cis), during which process autophagy was involved. FAM134B, the first detected endoplasmic reticulum autophagy (ER-phagy) receptor, plays an important part in the dynamic remodelling of the ER, the mutation of which affects sensory and autonomic neurons. However whether FAM134B-mediated ER-phagy involved in Cis-induced SGN damage or not was unknown. The present study was designed to determine whether FAM134B is expressed in SGNs of C57BL/6 mice and, if so, to explore the potential function of FAM134B in Cis-induced SGN damage in vitro . Methods Middle turns of neonatal murine cochleae were cultured and treated with 30 μM Cis in vitro . The distribution of FAM134B, morphological changes of SGNs, and the colocalization of ER segments with lysosomes were measured by immunofluorescence (IF). Apoptosis was measured by TUNEL staining. The expression of FAM134B, proteins associated with ER stress, autophagy and apoptosis was measured by western blot. The reactive oxygen specie (ROS) levels were evaluated by MitoSOX Red and 2′,7′-Dchlorodihydrofluorescein diacetate (DCFH-DA) probe. Anc80- Fam134b shRNA was used to knockdown the expression of FAM134B in SGNs. Results We first found the expression of FAM134B in the cytoplasm of SGNs, especially in the fourth postnatal day mice. Results showed decreases in the number of SGNs and FAM134B expression, as well as increases of ROS level, ER stress, ER-phagy, and apoptosis after Cis stimulus. Inhibiting autophagy increased the expression of FAM134B, and aggravated Cis-induced SGN damage, while the opposite changes were observed when autophagy was activated. Additionally, co-treatment with the N-Acetyl-L-Cysteine (NAC), ROS scavenger, alleviated Cis-induced ER stress, ER-phagy, and apoptosis. What’s more, knockdown the expression of FAM134B in SGNs made SGNs more vulnerable to cisplatin-induced injury. Discussion The present study revealed the expression pattern of FAM134B in C57BL/6 murine SGNs for the first time. Moreover, our work further verified the protective function of FAM134B mediated by ER-phagy in Cis-induced SGN apoptosis, at least partially, correlated with the accumulation of ROS and induction of ER stress, though the detailed regulatory mechanism through which needs much more work to reveal.
Previous research has shown that co-fermentation of Pichia kudriavzevii EP1 (EP) and Lactobacillus sanfranciscensis enhances the aroma of sourbread. This study aimed to investigate the mechanism behind this phenomenon by focusing on the fermentation of dough using EP1 alone and analyzing its impact on sourbread. Various methods, including gas chromatography-mass spectrometry, non-targeted metabolomics, and proteomic analysis, were employed to study the effects. The results indicated that EP1 yeast had limited utilization of monosaccharides, leading to decreased levels of organic acids but increased levels of amino acids. Analysis of volatile organic compounds (VOCs) revealed a higher presence of hexanal, a compound responsible for the characteristic flavor. Metabolomic analysis further revealed that EP1 yeast produced more oligosaccharides, alcohol substances, and small molecular peptides. Additionally, the precursor substance hexanol, associated with hexanal production, exhibited higher levels. Proteomic analysis indicated upregulation of proteins involved in fatty acid metabolism, protein metabolism, and amino acid metabolism in the EP1 yeast, supporting the observed changes in metabolite levels. Furthermore, a significant increase in the expression of alcohol dehydrogenase, associated with hexanal synthesis, was identified. Changes in protein expression showed a negative correlation with sugar metabolism changes and a positive correlation with changes in organic acid and amino acid metabolism.
Abstract Ferroptosis, characterized by iron‐dependent lipid reactive oxygen species (ROS) accumulation, plays a pivotal role in cisplatin‐induced ototoxicity. Existing research has suggested that in cisplatin‐mediated damage to auditory cells and hearing loss, ferroptosis is partially implicated. 4‐Octyl itaconate (4‐OI), derived from itaconic acid, effectively permeates cell membranes, showcasing potent anti‐inflammatory as well as antioxidant effects in several disease models. Our study aimed to investigate the effect of 4‐OI on cisplatin‐induced ferroptosis and the underlying molecular mechanisms. The survival rates of HEI‐OC1 cells and mice cochlea hair cells were measured by CCK8 and immunofluorescence, respectively. The auditory brainstem response (ABR) audiometry was used to detect changes in hearing thresholds in mice before and after treatment. Levels of ROS were evaluated by DCFH‐DA. Real‐time PCR quantified inflammatory cytokines TNF‐α, IL‐6 and IL‐1β. Network Pharmacology and RNA sequencing (RNA‐seq) analysis of the potential mechanism of 4‐OI resistance to cisplatin‐induced ferroptosis. The expressions of ferroptosis‐related factors (GPX4, SLC7A11 and PTGS2) and important antioxidant factors (NRF2, HO‐1, GCLC and NQO1) were tested by real‐time PCR, Western blot and immunofluorescence. Results demonstrated cisplatin‐induced significant ROS and inflammatory factor release, reduced NRF2 expression, hindered nuclear translocation and activated ferroptosis. Pretreatment with 4‐OI exhibited anti‐inflammatory and antioxidant effects, along with resistance to ferroptosis, ultimately mitigating cisplatin‐induced cell loss. In the present study, we show that 4‐OI inhibits cisplatin‐induced ferroptosis possibly through activation of the NRF2/HO‐1 signalling pathway, thereby exerting a protective effect against cisplatin‐induced damage to auditory cells, and providing a new therapeutic strategy for cisplatin‐induced hearing loss.
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