Mycoproteins and their health-promoting properties: Fusarium species and beyond.

Filamentous fungal mycoproteins have gained increasing attention as sustainable alternatives to animal and plant-based proteins. This comprehensive review summarizes the nutritional characteristics, toxicological aspects, and health-promoting effects of mycoproteins, focusing on those derived from filamentous fungi, notably Fusarium venenatum. Mycoproteins are characterized by their high protein content, and they have a superior essential amino acid profile compared to soybeans indicating excellent protein quality and benefits for human nutrition. Additionally, mycoproteins offer enhanced digestibility, further highlighting their suitability as a protein source. Furthermore, mycoproteins are rich in dietary fibers, which have been associated with health benefits, including protection against metabolic diseases. Moreover, their fatty acids profile, with significant proportions of polyunsaturated fatty acids and absence of cholesterol, distinguishes them from animal-derived proteins. In conclusion, the future of mycoproteins as a health-promoting protein alternative and the development of functional foods relies on several key aspects. These include improving the acceptance of mycoproteins, conducting further research into their mechanisms of action, addressing consumer preferences and perceptions, and ensuring safety and regulatory compliance. To fully unlock the potential of mycoproteins and meet the evolving needs of a health-conscious society, continuous interdisciplinary research, collaboration among stakeholders, and proactive engagement with consumers will be vital.

Optimized combination of Cervus nippon (Sika deer), Angelica (Dangui), and Rehmannia (Suk-jihwang) mitigates LPS-induced inflammation: exploring signaling pathways through plasma metabolomics.

This study aimed to determine the optimal combination of three anti-inflammatory materials [i.e., Cervus nippon Temminck (CT), Angelica gigas Nakai (AN), and Rehmannia glutinosa (RG)] for the strongest anti-inflammatory potential. Eighteen combinations of the three materials were tested in LPS-stimulated RAW264.7 cells via assessing nitric oxide (NO). The best combination from in vitro studies was administered to LPS-treated C57BL/6J mice for five days. Subsequently, plasma metabolites were profiled by bioinformatics analyses and validations. As results, 2, 20, and 50 µg/mL of CT, AN, and RG (TM) were the most effective combination suppressing inflammation. In mice, TM mitigated hepatic inflammatory markers. Similarly, the metabolomics indicated that TM may suppress NF-κB signaling pathway, thereby alleviating hepatic inflammation. TM also decreased systemic and hepatic pro-inflammatory cytokines. Collectively, we found the optimal combination of TM for mitigating inflammation; thus further studies on safety, mechanisms, and clinical models are warranted for human applications. Supplementary Information: The online version contains supplementary material available at 10.1007/s10068-023-01476-x.

An Amplicon-Based Application for the Whole-Genome Sequencing of GI-19 Lineage Infectious Bronchitis Virus Directly from Clinical Samples.

Infectious bronchitis virus (IBV) causes a highly contagious respiratory disease in chickens, leading to significant economic losses in the poultry industry worldwide. IBV exhibits a high mutation rate, resulting in the continuous emergence of new variants and strains. A complete genome analysis of IBV is crucial for understanding its characteristics. However, it is challenging to obtain whole-genome sequences from IBV-infected clinical samples due to the low abundance of IBV relative to the host genome. Here, we present a novel approach employing next-generation sequencing (NGS) to directly sequence the complete genome of IBV. Through in silico analysis, six primer pairs were designed to match various genotypes, including the GI-19 lineage of IBV. The primer sets successfully amplified six overlapping fragments by long-range PCR and the size of the amplicons ranged from 3.7 to 6.4 kb, resulting in full coverage of the IBV genome. Furthermore, utilizing Illumina sequencing, we obtained the complete genome sequences of two strains belonging to the GI-19 lineage (QX genotype) from clinical samples, with 100% coverage rates, over 1000 × mean depth coverage, and a high percentage of mapped reads to the reference genomes (96.63% and 97.66%). The reported method significantly improves the whole-genome sequencing of IBVs from clinical samples; thus, it can improve understanding of the epidemiology and evolution of IBVs.

The inhalation effect of Osmanthus fragrans var. Aurantiacus on physiological parameters in chronically stressed rats.

This study evaluated the effects of inhaling Osmanthus fragrans var. aurantiacus (OFA) extracts in Sprague-Dawley (SD) rats experiencing chronic stress. Rats were exposed to restraint stress or circadian disruption and were inhaled either distilled water or OFA extracts. Electronic nose (E-nose) analysis identified 35 volatile aromatic compounds (VACs) in OFA extracts. Chronic stress led to a decrease in body weight initially, serotonin concentration, and the weights of the liver, kidneys, and fat pads. Additionally, circadian disruption increased melatonin levels and decreased cholesterol concentrations. Inhalation of OFA increased dietary intake during the early phase and restored the tissue weights that have changed by chronic stress. Furthermore, it led to an increase in melatonin levels and changes in cholesterol levels. Taken together, our results indicate that OFA inhalation improves physiological changes caused by chronic stress through regulating dietary intake, restoring tissue weights, and modulating hormone and cholesterol levels.

Impact of roasting conditions on physicochemical, taste, volatile, and odor-active compound profiles of Coffea arabica L. (cv. Yellow Bourbon) using electronic sensors and GC-MS-O using a multivariate approach.

This study investigated the effects of roasting conditions on the physicochemical, taste, and volatile and odor-active compound (OAC) profiles of Coffea arabica L. At 150 ℃, roasting increased chlorogenic acid, total flavonoids, and caffeine concentrations. However, umami and sourness sensor decreased during the roasting process. At 210 ℃ roasting, total flavonoid and caffeine concentrations increased during the roasting process. Aldehydes, ketones, and sulfur-containing compounds dramatically increased during the roasting at 210 ℃ for 20 and 30 min in E-nose analysis. Pyrazines were mainly generated during the roasting at 210 ℃ for 20 and 30 min, and pyrazines showed the highest concentrations among all OACs in GC-olfactometry (GC-O) analysis. E-tongue data showed the separation of beans by roasting temperature. However, the E-nose and GC-O data showed the separation of beans by both roasting temperature and time via multivariate analysis. We identified similar results and patterns in the E-nose and GC-O analyses.

Foaming with Starch: Exploring Faba Bean Aquafaba as a Green Alternative.

The demand for sustainable and functional plant-based products is on the rise. Plant proteins and polysaccharides often provide emulsification and stabilization properties to food and food ingredients. Recently, chickpea cooking water, also known as aquafaba, has gained popularity as a substitute for egg whites in sauces, food foams, and baked goods due to its foaming and emulsifying capacities. This study presents a modified eco-friendly process to obtain process water from faba beans and isolate and characterize the foam-inducing components. The isolated material exhibits similar functional properties, such as foaming capacity, to aquafaba obtained by cooking pulses. To isolate the foam-inducing component, the faba bean process water was mixed with anhydrous ethanol, and a precipitated fraction was obtained. The precipitate was easily dissolved, and solutions prepared with the alcohol precipitate retained the foaming capacity of the original extract. Enzymatic treatment with α-amylase or protease resulted in reduced foaming capacity, indicating that both protein and carbohydrates contribute to the foaming capacity. The dried precipitate was found to be 23% protein (consisting of vicilin, α-legumin, and ß-legumin) and 77% carbohydrate (amylose). Future investigations into the chemical structure of this foam-inducing agent can inform the development of foaming agents through synthetic or enzymatic routes. Overall, this study provides a potential alternative to aquafaba and highlights the importance of exploring plant-based sources for functional ingredients in the food industry.

Effects of prebiotics, probiotics, and synbiotics on the infant gut microbiota and other health outcomes: A systematic review.

The primary aim of this review was to systematically evaluate the literature regarding the effect of pre-, pro-, or synbiotic supplementation in infant formula on the gastrointestinal microbiota. The Cochrane methodology for systematic reviews of randomized controlled trials (RCTs) was employed. Five databases were searched and 32 RCTs (2010-2021) were identified for inclusion: 20 prebiotic, 6 probiotic, and 6 synbiotic. The methods utilized to evaluate gastrointestinal microbiota varied across studies and included colony plating, fluorescence in situ hybridization, quantitative real-time polymerase chain reaction, or tagged sequencing of the 16S rRNA gene. Fecal Bifidobacterium levels increased with supplementation of prebiotics and synbiotics but not with probiotics alone. Probiotic and synbiotic supplementation generally increased fecal levels of the bacterial strain supplemented in the formula. Across all pre-, pro-, and synbiotic-supplemented formulas, results were inconsistent regarding fecal Clostridium levels. Fecal pH was lower with some prebiotic and synbiotic supplementation; however, no difference was seen with probiotics. Softer stools were often reported in infants supplemented with pre- and synbiotics, yet results were inconsistent for probiotic-supplemented formula. Limited evidence demonstrates that pre- and synbiotic supplementation increases fecal Bifidobacterium levels. Future studies utilizing comprehensive methodologies and additional studies in probiotics and synbiotics are warranted.

Chemical sensory investigation in green and roasted beans Coffea arabica L. (cv. Yellow Bourbon) by various brewing methods using electronic sensors.

This study investigated the effects of four brewing methods (cold and hot brew, espresso, and cezve) on the chemical sensory properties of green and roasted coffee beans (cv. Yellow Bourbon) extract. The caffeine and chlorogenic acid contents of the coffee were analyzed using HPLC. The taste and volatile aromatic compounds of coffee were analyzed using an electronic tongue and nose, respectively, and the results were analyzed using principal component analysis. For the taste components analyzed using the electronic tongue, the degree of separation was relatively large depending on the extraction method, and the degree of separation was larger depending on roasting for the volatile compounds analyzed using the electronic nose. Our findings provide basic data for the coffee industry. PRACTICAL APPLICATION: The use of an electronic sensor will provide flavor characteristics for four different types of coffee extracted from green beans and roasted beans. In this study, it was confirmed that the extraction method had a greater effect on the taste of coffee, and in the case of the volatile aromatic compounds of coffee, there was a large difference depending on the green beans and roasted beans. Therefore, our findings will provide data based on the sensory properties of coffee.

Antifatigue and Anti-Inflammatory Effects of Cervus elaphus L., Angelica gigas Nakai, and Astragalus membranaceus Bunge Complex Extracts in Physically Fatigued Mice.

Fatigue is a common complaint among people under stress, causing an array of negative effects on physical function. In this study, we investigated the antifatigue and anti-inflammatory effects of Cervus elaphus L., Angelica gigas Nakai, and Astragalus membranaceus Bunge complex extracts (CAA) using a treadmill stress test in animal models. The mice were administered various doses of CAA (50-200 mg/kg bw per day) once daily for 21 days. After exhaustive treadmill exercise, the running time of CAA-treated mice increased 1.5 times; fatigue-related biochemical parameters, including lactate dehydrogenase (∼30%), creatine kinase (∼20%), and proinflammatory cytokines interleukin (IL)-1ß (∼10%), and IL-6 (∼10%) in the serum and muscle tissue were downregulated compared with those in exercised control mice. This study provides strong evidence for the prevention of CAA-induced inflammatory incidences mediated by the blockade of nuclear factor-κB activation. Collectively, our results indicate that CAA can alleviate symptoms of fatigue in mice as an effective anti-inflammatory agent.

Canavalia gladiata Pod Extract Mitigates Ovalbumin-Induced Asthma Onset in Male BALB/c Mice via Suppression of MAPK.

Asthma is one of the most common inflammatory diseases of the lung worldwide. There has been considerable progress in recent studies to treat and prevent allergic asthma, however, various side effects are still observed in clinical practice. Six-week-old male BALB/c mice were orally administered with either sword bean pod extracts (SBP; 100 or 300 mg/kg) or dexamethasone (DEX; 5 mg/kg) once daily over 3 weeks, followed by ovalbumin sensitization (OVA/Alum.; intraperitoneal administration, 50 µg/2 mg/per mouse). Scoring of lung inflammation was performed to observe pathological changes in response to SBP treatment compared to OVA/Alum.-induced lung injury. Additionally, inflammatory cytokines were quantified in serum, bronchoalveolar lavage fluid (BALF), and lung tissue using ELISA and Western blot analyses. SBP treatment significantly reduced the infiltration of inflammatory cells, and release of histamine, immunoglobulin E, and leukotriene in serum and BALF. Moreover, the therapeutic effect of SBP was also assessed to analyze the inflammatory changes in the lung tissues. SBP markedly suppressed the activation of the MAPK signaling pathway and the expression of key inflammatory proteins (e.g., TNF-α) and Th2 type cytokines (IL-5 and IL-13). SBP was effective in ameliorating the allergic inflammation against OVA/Alum.-induced asthma by suppressing pulmonary inflammation.

Inhalation of low-dose basil (Ocimum basilicum) essential oil improved cardiovascular health and plasma lipid markers in high fat diet-induced obese rats.

This study investigated the antiobesogenic effects of the inhalation of volatile compounds derived from basil essential oil (BEO) in high fat diet-induced obese rats. A total of 47 volatile compounds were identified in BEO using gas chromatography-mass spectrometry. Major volatile compounds identified by olfactory testing include linalool oxide, linalool, 1-menthene, and carvone. White adipose tissue significantly decreased in the rats that inhaled 0.3% BEO (more than +10%) compared to the control. Plasma marker analysis showed increased high-density lipoprotein-cholesterol (ca. double fold) and decreased low-density lipoprotein-cholesterol (more than -30%) levels in inhaled 1% BEO group compared to the control. Leptin significantly decreased in the 0.3 and 1% BEO groups (more than -70 and -85%, respectively). Last, systolic blood pressure at week 12 was significantly lower in inhaled 1% BEO group (more than -15%) compared to the control. The results of this study suggest that BEO inhalation may be effective in managing plasma lipid markers (cholesterols and leptin) and possibly metabolic disorders such as obesity. Practical Application: Changes in metabolic health markers, which are effected by inhalation of volatiles in basil (Ocimum basilicum) essential oil, will provide physiological variations in vivo to the public. In this study, the opposite effects were identified between 0.3% and 1% inhalation, respectively. Therefore, our findings will provide optimized and useful guidance for inhalation of basil essential oil.

Acrolein, an environmental toxicant and its applications to in vivo and in vitro atherosclerosis models: An update.

Cardiovascular disease, the foremost cause of death worldwide, is an overarching disease term that encompasses a number of disorders involving the heart and circulatory system, including atherosclerosis. Atherosclerosis is a primary cause of cardiovascular diseases and is caused by buildup of plaque and narrowing of blood vessels. Epidemiological studies have suggested that environmental pollutants are implicated in atherosclerosis disease progression. Among many environmental pollutants, acrolein (Acr) is an abundant reactive aldehyde and is ubiquitously present in cigarette smoke as well as food products (e.g., overheated oils and wine). Despite its ubiquitous presence and potential impact on the etiology of cardiovascular disease, a limited consensus has been made in regard to Acr exposure conditions to induce atherosclerosis in vivo. This mini-review summarizes in vivo atherosclerosis models using Acr to investigate biochemical and phenotypic changes related to atherosclerosis and in vitro mechanistic studies involving Acr and atherosclerosis.

Olfactory Stimulation with Volatile Aroma Compounds of Basil (Ocimum basilicum L.) Essential Oil and Linalool Ameliorates White Fat Accumulation and Dyslipidemia in Chronically Stressed Rats.

We explored the physiological effects of inhaling basil essential oil (BEO) and/or linalool and identified odor-active aroma compounds in BEO using gas chromatography/mass spectrometry (GC-MS) and GC-olfactometry (GC-O). Linalool was identified as the major volatile compound in BEO. Three groups of rats were administered BEO and linalool via inhalation, while rats in the control group were not. Inhalation of BEO for 20 min only reduced the total weight gain (190.67 ± 2.52 g) and increased the forced swimming time (47.33 ± 14.84 s) compared with the control group (219.67 ± 2.08 g, 8.33 ± 5.13 s). Inhalation of BEO for 5 min (392 ± 21 beats/min) only reduced the pulse compared with the control group (420 ± 19 beats/min). Inhalation of linalool only reduced the weight of white adipose tissue (5.75 ± 0.61 g). The levels of stress-related hormones were not significantly different among the groups. The total cholesterol and triglyceride levels decreased after inhalation of BEO for 20 min (by more than -10% and -15%, respectively). Low-density lipoprotein cholesterol levels were lowered (by more than -10%) by the inhalation of BEO and linalool, regardless of the inhalation time. In particular, BEO inhalation for 20 min was associated with the lowest level of low-density lipoprotein cholesterol (53.94 ± 2.72 mg/dL). High-density lipoprotein cholesterol levels increased after inhalation of BEO (by more than +15%). The atherogenic index and cardiac risk factors were suppressed by BEO inhalation. Animals exposed to BEO and linalool had no significant differences in hepatotoxicity. These data suggest that the inhalation of BEO and linalool may ameliorate cardiovascular and lipid dysfunctions. These effects should be explored further for clinical applications.

Citrus junos Tanaka Peel Extract and Its Bioactive Naringin Reduce Fine Dust-Induced Respiratory Injury Markers in BALB/c Male Mice.

Particulate matter (PM) 10 refers to fine dust with a diameter of less than 10 µm and induces apoptosis and inflammatory responses through oxidative stress. Citrus junos Tanaka is a citrus fruit and contains bioactive flavonoids including naringin. In the present study, we aimed to identify the preventive effect of Citrus junos Tanaka peel extract (CPE) against PM10-induced lung injury. As a proof of concept, NCI-H460 cells were treated with CPE (800 µg/mL, 12 h) in conjunction with PM10 to examine intracellular antioxidative capacity in the pulmonary system. In an in vivo model, male BALB/c mice (n = 8/group) were randomly assigned into five groups: NEG (saline-treated), POS (PM10 only), NAR (PM10 + naringin, 100 mg/kg), CPL (PM10 + CPE low, 100 mg/kg), and CPH (PM10 + CPE high, 400 mg/kg). Intervention groups received dietary supplementations for 7 days followed by PM10 exposure (100 mg/kg, intranasal instillation). Compared to the NEG, the CPE decreased to 22% of the ROS generation and significantly increased cell viability in vitro. The histological assessments confirmed that pulmonary damages were alleviated in the PM10 + CPL group compared to the POS. Pro-inflammatory cytokines and NF-κB/apoptosis signaling-related markers were decreased in the PM10 + CPL group compared to the POS. These results indicated that CPE showed promising efficacy in preventing pulmonary injuries in vivo. Such protection can be explained by the anti-oxidative capacity of CPE, likely due to its bioactives, including naringin (7.74 mg/g CPE). Follow-up human intervention, as well as population-level studies, will further shed light on the preventive efficacy of CPE against pulmonary damage in humans.

Olfactory Stimulation by Fennel (Foeniculum vulgare Mill.) Essential Oil Improves Lipid Metabolism and Metabolic Disorders in High Fat-Induced Obese Rats.

In this study, odor components were analyzed using gas chromatography/mass spectrometry (GC/MS) and solid-phase microextraction (SPME), and odor-active compounds (OACs) were identified using GC-olfactometry (GC-O). Among the volatile compounds identified through GC-O, p-anisaldehyde, limonene, estragole, anethole, and trans-anethole elicit the fennel odor. In particular, trans-anethole showed the highest odor intensity and content. Changes in body weight during the experimental period showed decreasing values of fennel essential oil (FEO)-inhaled groups, with both body fat and visceral fat showing decreased levels. An improvement in the body's lipid metabolism was observed, as indicated by the increased levels of cholesterol and triglycerides and decreased levels of insulin in the FEO-inhaled groups compared to group H. Furthermore, the reduction in systolic blood pressure and pulse through the inhalation of FEO was confirmed. Our results indicated that FEO inhalation affected certain lipid metabolisms and cardiovascular health, which are obesity-related dysfunction indicators. Accordingly, this study can provide basic research data for further research as to protective applications of FEO, as well as their volatile profiles.

Apiaceous vegetables protect against acrolein-induced pulmonary injuries through modulating hepatic detoxification and inflammation in C57BL/6 male mice.

Acrolein (Acr) is a reactive aldehyde in the environment. Acr causes oxidative stress and a cascade of catalytic events and has, thereby, been associated with increased risk of pulmonary diseases. Whether apiaceous vegetables (API) consumption can prevent Acr-induced pulmonary toxicity has not yet been explored hence, we investigated the effects of API on Acr-induced pulmonary damages in C57BL/6J mice. The mice were assigned into either negative control [NEG group; American Institute of Nutrition (AIN)-93G diet only], positive control (POS group; AIN-93G+Acr) or API intervention group (API group; AIN-93G+21% API+Acr). After 1 week of dietary intervention, the POS and API mice were exposed to Acr (10 µmol/kg body weight/day) for 5 days. During the exposure period, assigned diets remained the same. Prominent indicators lung of toxicity of POS mice were found, including mucus accumulation, macrophage infiltration, and hemorrhage, all of which were ameliorated by the API. Serum and lung inflammation markers, such as a tumor necrosis factor alpha were also increased by Acr while reduced by API. In the liver, API upregulated expression of glutathione S-transferases, which enhanced the metabolism of Acr into water-soluble 3-hydroxypropyl mercapturic acid for excretion. This is consistent with observed reductions in serum Acr-protein adducts. Taken together, our results suggest that API may provide protection against Acr-induced pulmonary damages and inflammation via enhancement of the hepatic detoxification of Acr.

Detection of a novel porcine circovirus 4 in Korean pig herds using a loop-mediated isothermal amplification assay.

A novel porcine circovirus 4 has been recently identified in China and Korea. A sensitive and specific diagnostic method is urgently required to detect the virus in field samples. We developed a loop-mediated isothermal amplification (LAMP) the assay for the visual detection of PCV4 and evaluated its sensitivity, specificity, and applicability in clinical samples. This assay's results can be directly visualized by the naked eye using hydroxynaphthol blue after incubation for 40 min at 64 °C. The assay specifically amplified PCV4 DNA and no other viral nucleic acids. The sensitivity of the assay was <50 DNA copies/reaction, which was 10 times more sensitive than conventional polymerase chain reaction (cPCR) and comparable to real-time PCR (qPCR). Clinical evaluation revealed that the PCV4 detection rate in individual pig samples and at the farm level was 39.3 % (57/145) and 45.7 % (32/70), respectively, which were higher than cPCR (46 samples, 24 farms) and qPCR (52 samples, 29 farms) results. Cumulatively, owing to the advantages of high sensitivity and specificity, direct visual monitoring of the results, no possibility for cross-contamination, and being a low-cost equipment, the developed LAMP assay will be a valuable tool for the detection of the novel PCV4 in clinical samples, even in resource-limited laboratories.

A simple colorimetric detection of porcine epidemic diarrhea virus by reverse transcription loop-mediated isothermal amplification assay using hydroxynaphthol blue metal indicator.

A simple reverse transcription loop-mediated isothermal amplification combined with visual detection method (vRT-LAMP) assay was developed for rapid and specific detection of porcine epidemic diarrhea virus (PEDV) in this study, which overcomes the shortcomings of previously described RT-LAMP assays that require additional detection steps or pose a risk of cross-contamination. The assay results can be directly detected by the naked eye using hydroxynaphthol blue after incubating for 40 min at 62 °C. The assay specifically amplified PEDV RNA and no other viral nucleic acids. The limit of detection of the assay was less than 50 RNA copies per reaction, which was 100 times more sensitive than conventional reverse transcription polymerase chain reaction (RT-PCR) and comparable to real-time RT-PCR (RRT-PCR). In the clinical evaluation, the PEDV detection rate of vRT-LAMP was higher than that of RRT-PCR, showing 99 % concordance, with a kappa value (95 % confidence interval) of 0.97 (0.93-1.01). Considering the advantages of high sensitivity and specificity, simple and direct visual monitoring of the results, no possibility for cross-contamination, and being able to be used as low-cost equipment, the developed vRT-LAMP assay will be a valuable tool for detecting PEDV from clinical samples, even in resource-limited laboratories.

Fermentation of Chestnut (Catanea crenata Sieb) Inner Shell Enhances Anti-Obesity Effects in 3T3-L1 and C3H10T1/2 Adipocytes.

Chestnut inner shell (CIS) is rich in phenols and flavonoids such as gallic acid and ellagic acid, which are known to exhibit effective antioxidant and anti-obesity properties. Fermentation using lactic acid bacteria can enhance the physiological activity by increasing the contents of such functional ingredients. In this study, we evaluated the anti-obesity effects of a CIS extract subjected to a fermentation process (fermented CIS [FCIS]). Treatment with CIS and FCIS extracts (125, 250, and 500 µg/mL) increased cell viability and did not induce apoptosis, indicating no toxicity. The extract suppressed the gene expression of adipogenic factors, peroxisome proliferation-activated receptor gamma, CCAAT/enhancer binding protein (C/EBP) alpha, and C/EBP beta (by 7.75% and 67.59%, 21.41% and 66.27% in 500 µg/mL, respectively), and consequently suppressed the expression of downstream lipogenic factors such as fatty acid synthase, stearoyl CoA desaturase-1, citrate synthase, and ATP citrate lyase. The expression of factors involved in fat catabolism and ß-oxidation increased in a dose-dependent manner, thereby preventing fat accumulation. This observation was consistent with the significant decrease in the staining intensity for lipid droplets, which indicated that lipid accumulation was decreased by 15.46% and 29.44% in 3T3L-1 and 27.01% and 46.68% in C3H10T1/2. Together, these results demonstrate the higher anti-obesity effects of FCIS extract than that of CIS extract, indicating the potential applicability of FCIS as an effective natural raw material to curb obesity.

The role of microRNA-33 as a key regulator in hepatic lipogenesis signaling and a potential serological biomarker for NAFLD with excessive dietary fructose consumption in C57BL/6N mice.

Limited studies reported mechanisms by which microRNAs (miRNA) are interlinked in the etiology of fructose-induced non-alcoholic fatty liver disease (NAFLD). Here, we aimed to investigate the significance of miRNAs in fructose-induced NAFLD pathogenesis through unbiased approaches. In experiment I, C57BL/6N mice were fed either water or 34% fructose for six weeks ad libitum. In experiment II, time course effects of fructose intervention were monitored using the same conditions; mice were killed at the baseline, fourth, and sixth weeks. Bioinformatic analyses for hepatic proteomics revealed that SREBP1 is the most significant upstream regulator influenced by fructose; miR-33-5p (miR-33) was identified as the key miRNA responsible for SREBP1 regulation upon fructose intake, which was validated by in vitro transfection assay. In experiment II, we confirmed that the longer mice consumed fructose, the more severe liver injury markers (e.g., serum AST) appeared. Moreover, hepatic Srebp1 mRNA expression was increased depending upon the duration of fructose consumption. Hepatic miR-33 was time-dependently decreased by fructose while serum miR-33 expression was increased; these observations indicated that miR-33 from the liver might be released upon cell damage. Finally we observed that fructose-induced ferroptosis might be a cause of liver toxicity, resulting from oxidative damage. Collectively, our findings suggest that fructose-induced oxidative damage induces ferroptosis, and miR-33 could be used as a serological biomarker of fructose-induced NAFLD.