The Hydrolytic Peptides of Soybean Protein Induce Cell Cycle Arrest and Apoptosis on Human Oral Cancer Cell Line HSC-3.

Protein hydrolysates from various sources, including tuna cooking juice, soy protein isolate, sodium caseinate, wheat gluten and skin gelatin from porcine, tilapia, halibut and milkfish were analyzed to screen their antiproliferative activities against the human oral squamous carcinoma cell line, HSC-3. The soy protein isolate was selected for further investigations based on its hydrolysates with bromelain (SB) and thermolysin (ST), showing the greatest inhibition of cell growth. The SB and ST hydrolysates showed antiproliferative activities up to 35.45-76.39% against HSC-3 cells at 72 h, and their IC50 values were 0.74 and 0.60 mg/mL, respectively. SB and ST induced cell cycle arrest in the S phase through a pathway independent of p21 and p27 protein expression. Further, ST induced the apoptosis of HSC-3 cells by downregulating expression of Bcl-2, PARP, caspase 3 and caspase 9, but an upregulating expression of p53 and cleaved caspase 3. Unlike ST, SB may induce necrosis on HSC-3 cells. Thus, soybean hydrolysates may be a good source for providing antiproliferative peptides against HSC-3, while SB and ST may have the potential to be developed as functional foods.

Nobiletin Alleviates Ferroptosis-Associated Renal Injury, Inflammation, and Fibrosis in a Unilateral Ureteral Obstruction Mouse Model.

Nobiletin (Nob), a critical active flavonoid of citrus fruits, has received attention for its superior physical functions, which have shown to improve the progression of diseases. Chronic kidney disease (CKD) is recognized as a global health problem, and its mortality and morbidity rates are worsened with an increased risk of accompanying disorders. In this study, we aimed to elucidate whether Nob treatment ameliorates kidney fibrosis and also to identify the potential signaling networks in a unilateral ureteral obstructive (UUO) mouse model, which was used to mimic the progression of CKD. Six-week-old C57BL/6J mice were orally treated with 50 mg/kg of Nob for 14 constitutive days after UUO surgery. We found that the administration of Nob diminished kidney fibrosis and the expression of EMT markers, ameliorated oxidative stress and ferroptosis-associated injury, and mitigated the inflammatory response in the kidneys of UUO mice. Our results suggested that Nob treatment has antiferroptosis, anti-inflammatory, and antifibrotic effects, improving the progression of CKD in UUO mice. Nob may serve as a potential therapeutic candidate for the improvement of progressive CKD in further studies.

Establishment of optimal QuEChERS conditions of various food matrices for rapid measurement of heterocyclic amines in various foods.

The optimal conditions for QuEChERS (quick, easy, cheap, effective, rugged and safe) with superior performance were established to rapidly extract 21 heterocyclic amines (HAs) from 9 categories of food matrices including meat and poultry, eggs, soy beans and products, composite foods, fish and seafood, grains, beer, dairy foods, and coffee. The QuEChERS conditions and the developed ultra-high performance liquid chromatography-tandem mass spectrometry analysis conditions were then applied to the determination of HAs in popular food products sold in the Taiwan market. The conditions comply with the food chemical analysis specifications of Taiwan Food and Drug Administration. Coffee products and braised products that require a longer cooking time contained relatively high content of HAs (mainly Harman and Norharman), and their consumption is relatively high resulting in relatively high intake of HAs from these products. The dietary intake of HAs in plant-based protein food products should also be of concern.

Recent advances in the analysis of polycyclic aromatic hydrocarbons in food and water.

Polycyclic aromatic hydrocarbons (PAHs), a class of harmful and persistent organic contaminant, are widely distributed in the environment and eventually accumulated in water and food. Also, they are formed in different varieties and varying amounts during processing of food depending on the food composition, cooking method and processing condition. According to the International Agency for Research on Cancer (IARC), various PAHs are classified under Group 1 to 3 category, with Group 1 designated as carcinogenic to humans, Group 2A as probable carcinogen, Group 2B as possible carcinogen and Group 3 as noncarcinogenic. Therefore, it is imperative to develop rapid and highly sensitive analytical methods for determination of PAHs in food and water. This article aims to overview the recent advances of various chromatographic methods as well as electrochemical and SERS-based optical sensing methods for analysis of PAHs in food and water. Initially, several conventional sample preparation methods along with the advanced extraction for isolation of PAHs were summarized, followed by reviewing various gas chromatographic methods coupled with various detection techniques for PAHs analysis in various food products including meat/meat products, seafood, oil, milk/milk products, baby foods, honey, vegetable, cocoa products, tea/coffee, juice, rice, flour, noodle and cake. In addition, high performance liquid chromatographic methods coupled with fluorescence, diode array or mass/tandem mass detection techniques as well as an emerging supercritical fluid chromatographic technique employed for determination of PAHs in different food and water matrices were also overviewed. Finally, various electrochemical sensors and SERS-based optical sensors developed recently for onsite detection of PAHs were tabulated and discussed. Thus, this review article can provide a research update on chromatography and sensor-based analytical methods for PAH analysis as well as enable elucidation of research gaps for future studies.

Evaluation of optimal QuEChERS conditions of various food matrices for rapid determination of EU priority polycyclic aromatic hydrocarbons in various foods.

Optimal QuEChERS (quick, easy, cheap, effective, rugged and safe) conditions with good accuracy, repeatability and precision were established to rapidly extract the European Union (EU) priority polycyclic aromatic hydrocarbons (PAHs) from various food matrices (Category: Poultry and Meat, Fish and seafood, Grains, Soy beans and products, Root vegetables and Coffee). The QuEChERS conditions combined with the established high performance liquid chromatography-fluorescence detection conditions were used to rapidly determine the PAHs in 19 popular cooked foods in Taiwan and their corresponding original materials. These conditions also meet the EU and Taiwan Food and Drug Administration specifications. Charcoal grilled, gas stove grilled and smoked foods had higher PAHs contents, while fried and electric oven-baked/baked foods had lower PAHs contents. In addition to the effects of cooking methods, the contamination of original materials by PAHs in the environment should also have an important impact on the contents of PAHs in these cooked foods.

Implementation of food safety management systems that meets ISO 22000:2018 and HACCP: A case study of capsule biotechnology products of chaga mushroom.

In our study, a food safety management system was developed for a chaga mushroom biotechnology product manufacturer, with the purpose of meeting the quality demands of customers and enhancing the manufacturer's reputation. The study focused on identifying the potentially significant hazards present at each stage of the production process for chaga mushroom capsule products, and on ensuring that the biotechnology company in question has fully implemented ISO22000:2018 and the HACCP methodology. The results indicate that, in the 1 year following the implementation of ISO 22000:2018, there was a statistically significant drop (P < 0.05) in the coliforms level of the tested biotechnology product samples that started in the ninth month following implementation. The rapid screening of mycotoxin, heavy metal, and pesticide residue levels also increased monitoring intensity and strengthened the periodic rotation plan, enabling control over potential problems relating to raw materials and ensuring product quality. This finding reveals the importance and necessity of rapid screening for small- and medium-sized food industry enterprises. Furthermore, 1 year after the system's implementation, the defect rate for chaga products was also observed to have declined, and the number of process flow anomalies requiring correction was also found to have decreased significantly (P < 0.05), indicating that safety and quality levels were improving and stabilizing. If implemented over a longer period of time, the food safety management system's benefits would stand out even more, and significant improvements would be observed for more monitored items. PRACTICAL APPLICATION: Few studies have touched on food safety management systems (FSMSs) developed for capsule health food products. The enterprise examined in this study had actively worked to improve its quality system and meet its customers' needs through the implementation of the FSMSs.

Effects of operating conditions on biogas production in an anaerobic digestion system of the food and beverage industry.

BACKGROUND: Food residuals (FR) were anaerobically biotransformed to produce biogases (e.g. methane and hydrogen), and different pre-treatment conditions, including particle size, oil content, pH and salt content, were controlled in this study. The bio-solids of a municipal solid waste (MSW) from a wastewater treatment plant were added to assess its effect on anaerobic transformation efficiency and gas yields. RESULTS: The breaking of FR and the application of MSW were effective in enhancing the transformation efficiency and yield of biogases. The energy transfer efficiency value of the combined FRs used in this study was probably 23%. However, it can be very cost effective to apply arbitrary proportions to treat two types of FR in the anaerobic digestion tank of a wastewater treatment plant. It was also found that the alkalinity and pH value were two major parameters that controlled the success of the transformation. About 0.16-0.17 kg of alkalinity was needed during the anaerobic digestion of 1 kg dry FR, but this requirement was decreased by the treatment applying MSW. Olive oil had higher reducing rates when used as a substitute for heat-oxidized oil to study the effect of oil content on methylation. CONCLUSION: The conditions for anaerobic digestion established in this study were practical for the digestion of FR in wastewater treatment plants in Taiwan. However, we nonetheless found that it was cost effective to use arbitrary proportions for both types of FR and integrate the anaerobic digestion process used in wastewater treatment plants. © 2020 Society of Chemical Industry.

Comparison and establishment of appropriate methods to determine EU priority PAHs in charcoal-grilled chicken drumsticks with different treatments and their dietary risk assessments.

Three major polycyclic aromatic hydrocarbons (PAHs) analytical methods, gas chromatography-mass spectrometer (GC-MS), high performance liquid chromatography-photodiode-array detector (HPLC-PDA) and HPLC-fluorescence detector (FLD) were compared to analyze the European Union (EU) priority PAHs. In addition to the highest sensitivity, HPLC-FLD could be developed to rapidly determine the PAHs. A QuEChERS method was also established to rapidly extract the PAHs from chicken drumsticks. The method detection limits (0.004-0.25 ng/g), method quantification limits (0.01-0.75 ng/g), recoveries (67-114%), and coefficients of variations for intra- (1-15%) and inter- (1-21%) assays for the determination of the PAHs were in compliance with the EU and Taiwan Food and Drug Administration (TFDA) requirements. The PAHs contents in charcoal-grilled chicken drumsticks (without skin, with skin, and skin removal after processing) at different processing times were also determined by the developed methods. Although the risk assessment results for all samples showed a low level of concern, the presence of skin and long-term grilling had a greater impact on PAHs levels and dietary risks. Therefore, excessive processing should be avoided, and the skin of chicken drumsticks should be removed before and after charcoal-grilling to reduce the risk of PAHs intake.

Core food model of the Taiwan food supply for total diet study.

Following the core food (CF) models of the USA and France reported in the 2000s, this study presents the first oriental CF model, for total diet studies in Taiwan. First, we combined the four latest national dietary recall surveys from 2005 to 2012, data sets from the Nutrition and Health Survey in Taiwan (NAHSIT) covering all ages (1 to 65+ years old). In total, 3,098 NAHSIT food codes from 349,497 records of responses from 7,580 individuals were aggregated within a three-tiered scheme into 237 CFs in 54 subcategories and 16 categories. As with Western foods, Taiwanese foods were mainly aggregated by food type and major ingredient, processing method, cooking method or serving habit. In addition, we propose several categories typical to Taiwanese foods: (1) processed foods (e.g. rice paste, rice noodle, bean curd); (2) mixed foods (e.g. tomato fried with egg, Guangdong porridge) and stuffed staple foods (e.g. rice cake, zongzi); (3) Chinese Dim Sum (e.g. dumpling, salted bun); and (4) animal offal (e.g. pig liver, chicken kidney) and by-products (e.g. chicken head, pig foot). Systematic nomenclature and coding methods are also proposed. A computerised web model was developed using a graphical user interface (GUI) for automatic CF aggregation and inquiry of various consumption estimates according to the residing area, age group, and gender of consumers. Point estimates of weighted means were calculated for consumption rates (CR) based on the whole group (WG) or consumers only (CO). The CR and body weight (BW) of each individual were matched to calculate mean intake per unit body weight (g/kg bw/day). A general sample design procedure with the CF model is suggested for a total diet study (TDS) on target hazards. Strategies are also discussed for improving the quality of food matching in sample designs.

A study to evaluate the potential of an in silico approach for predicting dipeptidyl peptidase-IV inhibitory activity in vitro of protein hydrolysates.

A total of 294 edible protein sequences and 5 commercial proteases listed in the BIOPEP database were analyzed in silico. The frequency (A), a parameter in silico described previously, was examined further to calculating the ratio of truncated peptides with Xaa-proline and/or Xaa-alanine to all peptide fragments in a protein hydrolyzed with a protease, using the BIOPEP database. Then the in vitro DPP-IV inhibitory activity was determined using the same 15 protein and protease combinations to evaluate their relationship. The result shows that A values considering the number of Xaa-proline+Xaa-alanine exhibited a strong correlation with in vitro DPP-IV inhibition rates by Pearson's correlation analysis (r=0.6993; P<0.05). Therefore, the in silico approach is effective to predict DPP-IV inhibitory activities in vitro of protein hydrolysates.

In silico, in vitro and in vivo analyses of dipeptidyl peptidase IV inhibitory activity and the antidiabetic effect of sodium caseinate hydrolysate.

The frequency (A), a novel in silico parameter, was developed by calculating the ratio of the number of truncated peptides with Xaa-proline and Xaa-alanine to all peptide fragments from a protein hydrolyzed with a specific protease. The highest in vitro DPP-IV inhibitory activity (72.7%) was observed in the hydrolysate of sodium caseinate by bromelain (Cas/BRO), and the constituent proteins of bovine casein also had relatively high A values (0.10-0.17) with BRO hydrolysis. 1CBR (the <1 kDa fraction of Cas/BRO) showed the greatest in vitro DPP-IV inhibitory activity of 77.5% and was used for in vivo test by high-fat diet-fed and low-dose streptozotocin-induced diabetic rats. The daily administration of 1CBR for 6 weeks was effective to improve glycaemic control in diabetic rats. The results indicate that the novel in silico method has the potential as a screening tool to predict dietary proteins to generate DPP-IV inhibitory and antidiabetic peptides.

Isolation of prolyl endopeptidase inhibitory peptides from a sodium caseinate hydrolysate.

Prolyl endopeptidase (PEP) has been associated with neurodegenerative disorders, and the PEP inhibitors can restore the memory loss caused by amnesic compounds. In this study, we investigated the PEP inhibitory activity of the enzymatic hydrolysates from various food protein sources, and isolated and identified the PEP inhibitory peptides. The hydrolysate obtained from sodium caseinate using bromelain (SC/BML) displayed the highest inhibitory activity of 86.8% at 5 mg mL(-1) in the present study, and its IC50 value against PEP was 0.77 mg mL(-1). The F-5 fraction by RP-HPLC (reversed-phase high performance liquid chromatography) from SC/BML showed the highest PEP inhibition rate of 88.4%, and 9 peptide sequences were identified. The synthetic peptides (1245.63-1787.94 Da) showed dose-dependent inhibition effects on PEP as competitive inhibitors with IC50 values between 29.8 and 650.5 µM. The results suggest that the peptides derived from sodium caseinate have the potential to be PEP inhibitors.

The development of bioactive peptides from dietary proteins as a dipeptidyl peptidase IV inhibitor for the management of type 2 diabetes.

One of the new approaches to the management of type 2 diabetes mellitus (T2DM) consists of orally administered dipeptidyl peptidase-IV (DPP-IV) inhibitors. These synthetic drug inhibitors are reported to have some side effects and that subsequently limits their applications. There is a growing interest to develop natural DPP-IV inhibitors that will be potent without undesirable side effects. Many in vitro and some in vivo studies have highlighted the potential of food-derived peptides functioning as effective DPPIV inhibitors. Bioactive peptides within original food-derived proteins are inactive but can be activated by being released during food processing (by enzymatic hydrolysis or fermentation) or during gastrointestinal digestion. Hence, the utilization of computer-aided techniques as screening tools may be helpful in predicting the potential of food proteins as precursors of DPP-IV inhibitory peptides. This paper reviews the current literature on DPP-IV inhibitory peptides, focusing on their in vitro activity and in vivo antidiabetic effects. In addition, the feasibility of various in silico approaches is also summarized in this review.

Dipeptidyl-peptidase IV inhibitory activity of peptides derived from tuna cooking juice hydrolysates.

The in vitro DPP-IV inhibitory activity of isolated peptides from of tuna cooking juice hydrolyzed by Protease XXIII (PR) and orientase (OR) was determined. The results showed that the peptide fractions with the molecular weight over 1,422 Da possessed the greatest DPP-IV inhibitory activity. The amino acid sequences of the three peptides isolated from PR and OR hydrolysates were identified by MALDI-TOF/TOF MS/MS, and they were Pro-Gly-Val-Gly-Gly-Pro-Leu-Gly-Pro-Ile-Gly-Pro-Cys-Tyr-Glu (1412.7 Da), Cys-Ala-Tyr-Gln-Trp-Gln-Arg-Pro-Val-Asp-Arg-Ile-Arg (1690.8 Da) and Pro-Ala-Cys-Gly-Gly-Phe-Try-Ile-Ser-Gly-Arg-Pro-Gly (1304.6 Da), while they showed the dose-dependent inhibition effect of DPP-IV with IC(50) values of 116.1, 78.0 and 96.4 µM, respectively. In vitro simulated gastrointestinal digestion retained or even improved the DPP-IV inhibitory activities of the three peptides. The results suggest that tuna cooking juice would be a good precursor of DPP-IV inhibitor, and the DPP-IV inhibitory peptides can successfully passed through the digestive tract.

Peptides derived from atlantic salmon skin gelatin as dipeptidyl-peptidase IV inhibitors.

The dipeptidyl-peptidase IV (DPP-IV)-inhibitory activity of peptides derived from Atlantic salmon skin gelatin hydrolyzed by alcalase (ALA), bromelain (BRO), and Flavourzyme (FLA) was determined. The FLA hydrolysate with the enzyme/substrate ratio of 6% showed the greatest DPP-IV-inhibitory activity. The hydrolysate was fractionated by ultrafiltration with 1 and 2.5 kDa cutoff membranes, and the <1 kDa fraction had the highest DPP-IV-inhibitory activity with an IC(50) value of 1.35 mg/mL. The F-1 fraction further isolated by HPLC showed the IC(50) value against DPP-IV of 57.3 µg/mL, and the peptide sequences were identified as Gly-Pro-Ala-Glu (372.4 Da) and Gly-Pro-Gly-Ala (300.4 Da). The synthetic peptides showed dose-dependent inhibition effects on DPP-IV with IC(50) values of 49.6 and 41.9 µM, respectively. The results suggest that the peptides derived from Atlantic salmon skin gelatin would be beneficial ingredients for functional foods or pharmaceuticals against type 2 diabetes.

Effect of different high pressure treatments on shucking, biochemical, physical and sensory characteristics of oysters to elaborate a traditional Taiwanese oyster omelette.

BACKGROUND: Whole oysters (Crassostrea gigas) were processed using high-pressure (HP) treatment (150-300 MPa) to determine their shucking and biochemical properties. Subsequently, HP-treated oysters were cooked at 160 degrees C for 90 s, as when preparing the oyster omelette dish, to evaluate their physical and sensory characteristics as compared to raw oysters. RESULTS: The treatments of 250 and 300 MPa for 2 min and 0 min, respectively, resulted in 100% release. The pH of HP-treated oysters increased slightly from 6.50 to 6.82, and the moisture contents of the HP-treated oysters with or without further cooking were all higher than those of the control. The brightness, yellowness and cutting strength of HP-treated oysters with further cooking changed insignificantly, while the redness decreased compared to the control. Sensory evaluation showed that oysters treated at 250 and 300 MPa oysters after cooking received higher quality scores than the control. CONCLUSIONS: HP processing at 250 and 300 MPa proved to be a good method for oyster shucking. The HP-treated oysters cooked in the oyster omelette are acceptable to consumers. Overall, the application of HP as a processing method to improve the quality and acceptability of oysters and their related products would be possible.

Comparative study on histological structures of the vitelline membrane of hen and duck egg observed by cryo-scanning electron microscopy.

The histological structures of the vitelline membranes (VM) of hen and duck eggs were observed by cryo-scanning electron microscopy (cryo-SEM), and the chemical characteristics were also compared. The outer layer surface (OLS) of duck egg VM showed networks constructed by fibrils and sheets (0.1-5.2 microm in width), and that of hen egg presented networks formed only by sheets (2-6 microm in width). Thicker fibrils (0.5-1.5 microm in width) with different arrangement were observed on the inner layer surface (ILS) of duck egg VM as compared to those (0.3-0.7 microm in width) of hen egg VM. Upon separation, the outer surface of the outer layer (OSOL) and the inner surface of the inner layer (ISIL) of hen and duck egg VMs were quite similar to fresh VM except that the OSOL of duck egg VM showed networks constructed only by sheets. Thin fibrils interlaced above a bumpy or flat structure were observed at the exposed surface of the outer layer (ESOL) of hen and duck egg VMs. The exposed surfaces of inner layers (ESIL) of hen and duck egg VMs showed similar structures of fibrils, which joined, branched, and ran in straight lines for long distances up to 30 microm; however, the widths of the fibrils shown in ESOL and ESIL of duck egg VM were 0.1 and 0.7-1.4 microm, respectively, and were greater than those (<0.1 and 0.5-0.8 microm) of hen egg VM. The continuous membranes of both hen and duck egg VMs were still attached to the outer layers when separated. The content of protein, the major component of VM, was higher in duck egg VM (88.6%) than in hen egg VM (81.6%). Four and six major SDS-soluble protein patterns with distinct localization were observed in hen and duck egg VMs, respectively. Overall, the different histological structures of hen and duck egg VMs were suggested to be majorly attributable to the diverse protein components.

High-pressure treatment for shelf-life extension and quality improvement of oysters cooked in a traditional Taiwanese oyster omelet.

Whole oysters were processed using high-pressure (HP) treatment at 250 and 300 MPa for 0 to 10 min and stored at 4 degrees Celsius for up to 28 days. HP-treated oysters and untreated oysters were evaluated for lipid oxidation, growth of microorganisms, and sensory characteristics after cooking at 160 degrees Celsius for 90 s. Microbial counts after HP treatment revealed that the bacterial load was initially reduced at all pressures. HP-treated oysters had significantly higher pH and moisture (P < 0.05) relative to control (untreated) oysters during storage. HP treatment increased lipid oxidation with unpleasant odor during storage compared with the control. HP treatment decreased redness but did not significantly affect the brightness and yellowness of cooked oysters. From tests of mechanical properties, 300 MPa-treated oysters after cooking had significantly increased toughness as measured by cutting force. HP-treated oysters after cooking received higher quality scores than did the control during the storage trial. Results indicated that 300 MPa for 2 min is the optimum HP treatment that results in oysters most acceptable for oyster omelets during storage at 4 degrees Celsius, and this treatment may extend the shelf life of these oysters to 21 days.

Effects of hydrostatic pressure/heat combinations on water uptake and gelatinization characteristics of japonica rice grains: a kinetic study.

The combination effects of pressure (200 to 500 MPa) and temperature (20, 40, and 50 degrees C) on the water uptake and gelatinization characteristics of japonica rice (Tainung 71) grains were investigated. Pressure greater than 200 MPa at all temperatures increased the moisture content and volume of rice grains; meanwhile, the increase content of rice grain volume showed a high correlation with that of moisture content (r(2)= 0.96). The highest degree of gelatinization of 73% was observed at 500 MPa and 50 degrees C for 120 min, while gelatinization did not occur at pressures below 300 MPa and temperatures of 20 and 40 degrees C. The rate of gelatinization followed the 1st-order kinetics at each temperature and pressure. The higher pressures and temperatures would result in higher values of rate constant k which could be correlated with both pressure and temperature by combining Arrhenius and Eyring models.

Histological structures of native and cooked yolks from duck egg observed by SEM and cryo-SEM.

A method was used to fix duck egg yolk while retaining its original sol structure to elucidate the fine structure of native yolk by using fixation with liquid nitrogen and cryo-scanning electron microscopy (cryo-SEM). Native yolk spheres showed a polyhedron shape with a diameter at approximately 50 to 100 µm and packed closely together. Furthermore, the interior microstructure of the native yolk spheres showed that a great amount of round globules ranging from 0.5 to 1.5 µm were embedded in a continuous phase with a lot of voids. After cooking, the sizes of the spheres were almost unchanged, and the continuous phase became a fibrous network structure observed by SEM with chemical fixation probably constituted of low density lipoprotein (LDL). The fine structure of the native yolk can be observed by cryo-SEM; however, the microstructure of yolk granules and plasma from cooked shell eggs can be observed by SEM with chemical fixation.