Comparative analysis of mozzarella cheeses fortified with whey protein hydrolysates, diverse in hydrolysis time and concentrations.

The objective of the present study was to improve the quality of mozzarella cheese using whey protein concentrates (WPCs) hydrolyzed for varying lengths of time (1 and 3 h). Four types of cheeses were made incorporating hydrolyzed WPCs in milk 3 and 6 % level and evaluated for nutritional, structural, and functional properties during 28 days storage at 4 °C. Whey protein hydrolysates (WPHs) incorporation increased protein, lactose, minerals, water-soluble-protein, non-protein-nitrogen. Mozzarella incorporated with WPHs hydrolyzed for 3 h had higher fat contents, favorable meltability and lower browning effect, stretchability, brittleness, springiness, and cohesiveness compared to mozzarella fortified with WPHs hydrolyzed for 1 h. The incorporation of hydrolyzed WPCs significantly influenced rheological and functional characteristics of mozzarella cheese. The cheeses made with hydrolyzed WPCs showed fewer changes in whiteness than control during storage. It was observed that both extent of hydrolysis and levels of WPHs incorporation had significant effect on the characteristics of mozzarella cheeses.

Cannabidiol mediates epidermal terminal differentiation and redox homeostasis through aryl hydrocarbon receptor (AhR)-dependent signaling.

BACKGROUND: Cannabidiol, a non-psychoactive phytocannabinoid, has antioxidant and anti-inflammatory activity in keratinocytes. However, the signaling pathway through which cannabidiol exerts its effect on keratinocytes or whether it can modulate keratinocyte differentiation has not been fully elucidated yet. OBJECTIVE: We investigated whether cannabidiol modulates epidermal differentiation and scavenges reactive oxygen species through the aryl hydrocarbon receptor (AhR) in keratinocytes and epidermal equivalents. METHODS: We investigated the cannabidiol-induced activation of AhR using AhR luciferase reporter assay, qRT-PCR, western blot, and immunofluorescence assays. We also analyzed whether keratinocyte differentiation and antioxidant activity are regulated by cannabidiol-induced AhR activation. RESULTS: In both keratinocytes and epidermal equivalents, cannabidiol increased both the mRNA and protein expression of filaggrin, involucrin, NRF2, and NQO1 and the mRNA expression of the AhR target genes, including CYP1A1 and aryl hydrocarbon receptor repressor. Additionally, cannabidiol showed antioxidant activity that was attenuated by AhR knockdown or co-administration with an AhR antagonist. Moreover, cannabidiol increased the ratio of OVOL1/OVOL2 mRNA expression, which is a downstream regulator of AhR that mediates epidermal differentiation. In addition to increased expression of barrier-related proteins, cannabidiol-treated epidermal equivalent showed a more prominent granular layer than the control epidermis. The increased granular layer by cannabidiol was suppressed by the AhR antagonist. CONCLUSION: Cannabidiol can be a modulator of the AhR-OVOL1-filaggrin axis and AhR-NRF2-NQO1 signaling, thus indicating a potential use of cannabidiol in skin barrier enhancement and reducing oxidative stress.

Alternative splicing variant of NRP/B promotes tumorigenesis of gastric cancer.

Gastrointestinal cancer is associated with a high mortality rate. Here, we report that the splice variant of NRP/B contributes to tumorigenic activity in highly malignant gastric cancer through dissociation from the tumor repressor, HDAC5. NRP/B mRNA expression is significantly higher in the human gastric cancer tissues than in the normal tissues. Further, high levels of both the NRP/B splice variant and Lgr5, but not the full-length protein, are found in highly tumorigenic gastric tumor cells, but not in non-tumorigenic cells. The loss of NRP/B markedly inhibits cell migration and invasion, which reduces tumor formation in vivo. Importantly, the inhibition of alternative splicing increases the levels of NRP/B-1 mRNA and protein in AGS cells. The ectopic expression of full-length NRP/B exhibits tumor-suppressive activity, whereas NRP/B-2 induces the noninvasive human gastric cancer cells tumorigenesis. The splice variant NRP/B-2 which loses the capacity to interact with tumor repressors promoted oncogenic activity, suggesting that the BTB/POZ domain in the N-terminus has a crucial role in the suppression of gastric cancer. Therefore, the regulation of alternative splicing of the NRP/B gene is a potential novel target for the treatment of gastrointestinal cancer. [BMB Reports 2022; 55(7): 348-353].

Structural and Functional Validation of a Full-Thickness Self-Assembled Skin Equivalent for Disease Modeling.

Recently, various types of in vitro-reconstructed 3D skin models have been developed for drug testing and disease modeling. Herein, we structurally and functionally validated a self-assembled reconstructed skin equivalent (RSE) and developed an IL-17a-induced in vitro psoriasis-like model using a self-assembled RSE. The tissue engineering approach was used to construct the self-assembled RSE. The dermal layer was generated using fibroblasts secreting their own ECM, and the epidermal layer was reconstructed by seeding keratinocytes on the dermal layer. To generate the psoriatic model, IL-17A was added to the culture medium during the air-liquid interface culture period. Self-assembled RSE resulted in a fully differentiated epidermal layer, a well-established basement membrane, and dermal collagen deposition. In addition, self-assembled RSE was tested for 20 reference chemicals according to the Performance Standard of OECD TG439 and showed overall sensitivity, specificity, and accuracy of 100%, 90%, and 95%, respectively. The IL-17a-treated psoriatic RSE model exhibited psoriatic epidermal characteristics, such as epidermal hyperproliferation, parakeratosis, and increased expression of KRT6, KRT17, hBD2, and S100A9. Thus, our results suggest that a self-assembled RSE that structurally and functionally mimics the human skin has a great potential for testing various drugs or cosmetic ingredients and modeling inflammatory skin diseases.

PFN1 Prevents Psoriasis Pathogenesis through IκBζ Regulation.

PFN1 is an actin-binding protein that regulates actin polymerization, cell proliferation, apoptosis, angiogenesis, and carcinogenesis. Its dysregulation has been reported in diverse pathologic diseases; however, the role of PFN1 in psoriasis has not yet been elucidated. In this study, we show that PFN1 expression is increased in both skin and serum of patients with psoriasis. PFN1 was markedly expressed in the epidermis of psoriatic lesions, and its expression positively correlated with psoriasis severity. IL-17A treatment of keratinocytes increased PFN1 expression, whereas TNF-α induced PFN1 expression and secretion. In addition, knockdown of PFN1 with short hairpin RNA resulted in an altered expression of psoriasis-associated inflammatory markers, HBD2, S100A7, S100A9, and Ki-67, and recombinant PFN1 suppressed the IL-17A‒induced inflammatory response in keratinocytes. Interestingly, recombinant PFN1 also suppressed IL-17A‒induced IκBζ, an important player in immune response in psoriasis. Collectively, our results show that PFN1 acts as a negative regulator of psoriatic inflammation through the suppression of IκBζ and that the balanced level of PFN1 is important for IκBζ regulation. Thus, the expression of PFN1 can be used as a biomarker for psoriasis severity, and it can be considered as a possible target for the treatment of psoriasis.

Quercetin intake, MATE1 polymorphism, and metabolic syndrome in Korean population: Hallym aging study.

Multidrug and toxic compound extrusion transporter-1 (MATE1) is a quercetin transporter. We examined the associations of quercetin intake and polymorphism of MATE1 in relation to metabolic syndrome (MetS) in Hallym Aging Study. Quercetin intake and the measurements for MetS were assessed in 2004. Six tagging single nucleotide polymorphisms (SNPs) at MATE1 gene were genotyped in 428 Korean adults in 2012. We found a lower prevalence of MetS with quercetin intake; compared to the lowest quartile, odds ratios (ORs, 95% confidence intervals; CIs) were 0.44 (0.24-0.84) for the 3rd quartile. Individuals with the minor allele of MATE1, rs2453589, tended to have a lower prevalence of MetS compared to those with the major allele (OR=0.69; CI=0.36-1.34). However, interactions between quercetin intake and six MATE1 polymorphisms in relation to MetS were not significant (p for interaction ≥0.37). In conclusion, intake of quercetin was associated with MetS in Korean populations.

Physicochemical and Microbiological Properties of Yogurt-cheese Manufactured with Ultrafiltrated Cow's Milk and Soy Milk Blends.

The objective of this study was to develop yogurt-cheese using cow's milk, ultrafiltrated cow's milk, and soy milk. The addition of soy milk and ultrafiltrated milk increased the amount of protein in the yogurt-cheese. Yogurt-cheeses were made using cheese base using 10% and 20% soy milk with raw and ultrafiltrated cow's milk, and stored at 4℃ during 2 wk. The yield of yogurt-cheeses made with added soy milk was decreased and the cutting point was delayed compared to yogurt-cheese made without soy milk. Yogurt-cheese made using ultrafiltrated cow's milk showed the highest yield. However, yogurt-cheese made with added soy milk had higher protein content and titratable acidity than yogurt-cheese made using raw and ultrafiltrated cow's milk. Fat and lactose contents in the yogurt-cheese made with added soy milk were lower. Yogurt-cheeses made with added soy milk contained several soy protein bands corresponding to the sizes of α2-, ß-, and κ-casein band. Yogurt-cheese made with added soy milk had similar elasticity to yogurt-cheese made without soy milk but had lower cohesiveness. There was no significant difference in the number of lactic acid bacteria in the different cheeses, as all had over 8.0 Log CFU/g. Considering these data and the fact that proteins and fats of vegetable origin with high biological value were observed as well as unsaturated fats, yogurt-cheese made with added soy milk can be considered to be a functional food.