The present study was conducted to investigate effects of rabbit meat extract on energy metabolism and muscle differentiation in C2C12 myotubes. Water extract of rabbit meat (10, 50, 100, and 200 μg/ml) was used to treat differentiated C2C12 cells. Reverse transcriptase polymerase chain reaction (RT-PCR) and western blot analysis were used to determine mRNA or protein levels of energy metabolism-related genes. Total adenosine triphosphate (ATP) content was also measured. Treatment with rabbit meat extract significantly increased expression levels of muscle differentiation markers (myogenin and myosin heavy chain) and mitochondrial biogenesis regulators (PGC1α, NRF1, and TFAM) in C2C12 myotubes compared to non-treated control. Additionally, rabbit meat extract activated phosphorylation of AMPK and acetyl-coA carboxylase (ACC). Rabbit meat extract significantly increased ATP contents in myotubes. These results suggest that rabbit meat extract has the potential to improve energy metabolism in skeletal muscles.

Background: The clinical application of canine mesenchymal stem cells (MSCs) necessitates efficient and safe culture methods to produce large quantities of cells. Traditionally, fetal bovine serum (FBS) has been used for MSC expansion, but it carries risks such as contamination and adverse immune responses. Methods: In this study, we investigate the efficacy and efficiency of canine allogeneic serum as an effective alternative to FBS for the in vitro culture of canine MSCs. We measured the population doubling time of canine MSCs in allogeneic serum conditions and utilized qRT-PCR, flowcytometric analysis, and cellular staining/color-metric assay for investigating its effects on cellular senescence during long-term culture and the expression of key pluripotency-related transcriptomes. Results: Our findings demonstrate that canine MSCs cultured with allogeneic serum exhibited enhanced proliferation rates, reduced cellular senescence, and lower apoptosis levels compared to those cultured with FBS. Additionally, the expression of key pluripotency-related transcription factors, including Oct4, Sox2, and Nanog, was increased in canine MSCs cultured with allogeneic serum. Conclusions: These results highlight the potential of canine allogeneic serum to provide a safer and more effective culture environment, supporting the large-scale expansion and maintenance of canine MSCs for clinical applications.

Periodontal disease (PD) is strongly linked to increased risk of oral squamous cell carcinoma (OSCC); however, the specific mechanism through which the development of PD and OSCC is simultaneously promoted remains unclear. This study explored the impact of periodontal pathogens on OSCC progression and the contribution of periodontal pathogen-stimulated OSCC to PD development. The expression of osteoclastogenesis-inducing factors was assessed using quantitative reverse transcription polymerase chain reaction analysis following stimulation of OSCC with lipopolysaccharide (LPS) derived from the periodontal pathogen Porphyromonas gingivalis (Pg), a pathogen commonly responsible for PD. The cell counting kit-8 assay was used to determine the effects of Pg-LPS on OSCC proliferation and drug resistance to cisplatin and 5-fluorouracil. The effects of conditioned medium (CM) derived from Pg-LPS–stimulated OSCC on osteoclastogenesis was evaluated using tartrate-resistant acid phosphatase (TRAP) staining on bone marrow-derived macrophages (BMMs). Pg-LPS administration in SCC-25 and YD-8 OSCC cell lines induced a significant increase in receptor activator of nuclear factor kappa-B ligand mRNA expression; however, it did not affect cell proliferation. Treatment with CM derived from Pg-LPS–stimulated SCC-25 or YD-8 cells markedly enhanced the formation of TRAP-positive multinucleated cells during osteoclast differentiation of BMMs. Altogether, these findings demonstrate that Pg-LPS–stimulated OSCC promoted osteoclastogenesis through a paracrine mechanism.

Health concerns related to particulate matter (PM) pollution are on the rise globally. This study investigates the effects of the main components of PM on human airway epithelial cells (Calu-3), focusing on three distinct types: PM10-bound PAHs (including Benzo[a]anthracene and Benzo[b]fluoranthene), PM10-bound trace elements (containing arsenic and lead), and PM2.5-bound ions (comprising sodium and calcium). Calu-3 cells were exposed to these PM components at concentrations ranging from 2 to 100 μg/mL. Unexposed Calu-3 cells exhibited a 60% increase in metabolic activity after 12 hours. In contrast, exposure to PM components resulted in significant reductions in cell viability, with PM10-bound PAHs and PM10-bound trace elements causing decreases of 54% and 55% respectively, and PM2.5-bound ions leading to a 63% reduction at 100 μg/mL. Additionally, there was found to be a notable rise in the expression of proinflammatory cytokines IL-8 and TNF-α. Specifically, IL-8 levels increased by 456%, and TNF-α levels rose by 660% after 12 hours of exposure to PM2.5-bound ions. These findings indicate that the size and composition of fine dust particles play a critical role in their cytotoxic effects, contributing to increased cell death, membrane damage, and necrosis in airway epithelial cells.

Obesity, a global health concern characterized by excessive fat accumulation, necessitates the discovery of anti-obesity compounds. Rottlerin, known for its anti-cancer effects as a mitochondrial uncoupler, has been a subject of interest. However, its impact on reducing intracellular lipid accumulation remains a gap in our understanding. This study aimed to fill this gap by dissecting the mechanism of rottlerin in 3T3-L1 adipocytes. We treated differentiated 3T3-L1 cells with 0-20 mM of rottlerin for 48 hours to assess its capability to induce lipid accumulation. Notably, we observed no cytotoxicity associated with the treatment of rottlerin up to 20 mM, indicating its safety at these concentrations. Lipid accumulation, measured by oil Red O, was downregulated dose-independently by rottlerin. We also found that key lipogenic enzymes, including SCD1 and DGAT1, were decreased. The transcription factor of lipogenic genes, SREBP1, was reduced by approximately 80% with rottlerin. LRP6, a crucial link between de novo lipogenesis mechanism reactions and Wnt signaling, was also degraded by around 70%. Interestingly, the downstream regulation of LRP6, b-catenin, and TCFL2 was diminished by rottlerin. Our data indicate that rottlerin alleviates adipocyte lipid accumulation by suppressing the LRP6/β-catenin/SREBP1c pathway. These findings underscore the potential of rottlerin as a safe nutraceutical for combating obesity.

Human bitter taste-sensing type 2 receptors (hTAS2Rs) are expressed in various human tissues and may be associated with various cell signaling pathways, cell progression, and cell physiology in each tissue. hTAS2Rs can be a potential drug target because it is also expressed in some cancer cells. Xanthorrhizol (XNT) has various biological activities, such as anticancer, antimicrobial, anti-inflammatory, and antioxidant. XNT produces a bitter taste, but the specific hTAS2R activated is unknown, and the hTAS2R-mediated effect of XNT on cancer cells has not been studied. This study discovered the target receptor of XNT among 25 hTAS2Rs and confirmed the possibility of the hTAS2R-mediated inhibition of cancer cell proliferation. XNT activated only one receptor, hTAS2R38 (EC50=1.606±0.021 g/mL), and its activity was inhibited by probenecid, a hTAS2R38 antagonist. When HepG2 and MCF-7 cells were treated with XNT or phenylthiocarbamide (PTC), a known hTAS2R38 agonist, both chemicals inhibited cancer cell proliferation. XNT targets the human bitter taste receptor TAS2R38 and inhibits the proliferation of HepG2 and MCF-7 cells mediated by TAS2R38. This suggests that TAS2R38 may be a new target for disease treatment and a potential new factor for drug development.

Inflammatory bowel disease (IBD) is a chronic condition characterized by continuous inflammation of the gastrointestinal tract that varies in intensity over time. IBD is caused by several factors including aberrant gut flora, immunological dysregulation, altered environmental conditions, and genetic variations. However, the pathogenesis of IBD remains unclear. Studies have indicated that an imbalance between T helper 17 (Th17) and regulatory T (Treg) cells contributes significantly to the development of IBD. Intestinal Tregs suppress inflammation and are critical for maintaining tissue homeostasis. Th17 cells are known to play an important role in the development and pathogenesis of IBD and provide non-inflammatory support for the integrity of the intestinal barrier against bacterial and fungal infections. Therefore, the Th17/Treg cell balance is crucial in the pathogenesis of IBD and gut integrity. The microenvironment of the intestinal mucosal immunity is regulated by the secretion of cytokines associated with Th17 cells and Tregs. Several studies have indicated that the gut bacteria contribute to the control of the immune milieu and play a key role in the regulation of Th17 cell development. Intestinal bacteria and cytokines control Th17 cell development. Th17 cells secrete cytokines that regulate the immune microenvironment in the gut mucosa. This review provides an overview of Th17 cells and examines the strategies for treating patients with IBD using Th17 cell-targeted drugs.

Sprout products, such as broccoli, alfalfa, and cabbage, have positive health effects. Thus far, sprout foods have attracted attention owing to their good bioavailability. In particular, young broccoli sprouts exhibit anti-inflammatory, antioxidant, and anti-cancer effects. They contain 100 times more chemoprotective substances than adult broccoli. This study examined the anti-inflammatory effects of freeze-dried young sprout broccoli (FD-YB) in vitro using RAW264.7 macrophage cells. The FDYB powder antioxidant ability test showed that the radical-scavenging activity and superoxide dismutase enzyme activity increased in a dose-dependent manner. In addition, FD-YB was not cytotoxic to RAW264.7 cells, and nitric oxide production decreased after the FD-YB treatment of lipopolysaccharide-stimulated RAW264.7 cells in a dose-dependent manner. Furthermore, FD-YB significantly decreased the expression of inflammation-related proteins (Cyclooxygenase-2, Inducible nitric oxide synthase, and Prostaglandin E Synthase 2) and cytokines (Tumor necrosis factor- and Interleukin-6). In conclusion, FD-YB can be a potential nutraceutical for preventing and regulating excessive immune responses during inflammation.

Background: Aflatoxin B1 (AFB1) is a toxic metabolite generated by Aspergillus species and is commonly detected during the processing and storage of food; it is considered a group I carcinogen. The hepatotoxic effects, diseases, and mechanisms induced by AFB1 owing to chronic or acute exposure are well documented; however, there is a lack of research on its effects on the intestine, which is a crucial organ in the digestive process. Dogs are often susceptible to chronic AFB1 exposure owing to lack of variation in their diet, unlike humans, thereby rendering them prone to its effects. Therefore, we investigated the effects of AFB1 on canine small intestinal epithelial primary cells (CSIc). Methods: We treated CSIc with various concentrations of AFB1 (0, 1.25, 2.5, 5, 10, 20, 40, and 80 μM) for 24 h and analyzed cell viability and transepithelial-transendothelial electrical resistance (TEER) value. Additionally, we analyzed the mRNA expression of tight junction-related genes (OCLN, CLDN3, TJP1, and MUC2), antioxidant-related genes (CAT and GPX1), and apoptosis-related genes (BCL2, Bax, and TP53). Results: We found a significant decrease in CSIc viability and TEER values after treatment with AFB1 at concentrations of 20 μM or higher. Quantitative polymerase chain reaction analysis indicated a downregulation of OCLN, CLDN3, and TJP1 in CSIc treated with 20 μM or higher concentrations of AFB1. Additionally, AFB1 treatment downregulated CAT , GPX1, and BCL2. Conclusions: Acute exposure of CSIc to AFB1 induces toxicity, and exposure to AFB1 above a certain threshold compromises the barrier integrity of CSIc.

Background: In healthy dentin conditions, odontoblasts have an important role such as protection from invasion of pathogens. In mammalian teeth, progenitors such as mesenchymal stem cells (MSCs) can migrate and differentiate into odontoblast-like cells, leading to the formation of reparative dentin. For differentiation using stem cells, it is crucial to provide conditions similar to the complex and intricate in vivo environment. The purpose of this study was to evaluate the potential of differentiation into odonto/ osteoblasts, and compare co-culture with/without epithelial cells. Methods: MSCs and epithelial cells were successfully isolated from dental tissues. We investigated the influences of epithelial cells on the differentiation process of dental pulp stem cells into odonto/osteoblasts using co-culture systems. The differentiation potential with/without epithelial cells was analyzed for the expression of specific markers and calcium contents. Results: Differentiated odonto/osteoblast derived from dental pulp tissue-derived mesenchymal stem cells with/without epithelial cells were evaluated by qRT-PCR, immunostaining, calcium content, and ALP staining. The expression of odonto/ osteoblast-specific markers, calcium content, and ALP staining intensity were significantly increased in differentiated cells. Moreover, the odonto/osteogenic differentiation capacity with epithelial cells co-culture was significantly higher than without epithelial cells co-culture. Conclusions: These results suggest that odonto/osteogenic differentiation co-cultured with epithelial cells has a more efficient application.

Probiotic lactic acid bacteria are live microorganisms that provide health benefits when administered in adequate amounts and may exhibit antiproliferative effects on various cancer cell lines, including colon cancer. This study investigates the cytotoxic effects of three Lactobacillus strains - Limosilactobacillus (L.) reuteri VA 102, Ligilactobacillus (L.) animalis VA 105, and Limosilactobacillus (L.) reuteri KCTC 3594 (ATCC 23272) - on mouse colon carcinoma cells (CT-26). Live cells, heat-killed cells, and cell-free supernatant (CFS) of Lactobacillus sp. were prepared and used to treat CT-26 cells at different concentrations. The cytotoxic effect was assessed using the MTT assay. The results indicated that the CFS of all strains significantly reduced the viability of CT-26 cells in a dose-dependent manner, with the VA 102 strain showing the most pronounced effect. Heat-killed cells of L. reuteri VA 102 and L. reuteri KCTC 3594 (ATCC 23272) also reduced cell viability. These findings suggest the potential anticancer properties of these Lactobacillus strains and indicate that CFS and heat-killed cells may offer a safer and more effective alternative to live bacteria for therapeutic applications. Our study contributes to the understanding of the potential of Lactobacillus strains, particularly L. reuteri VA 102, L. reuteri KCTC 3594 (ATCC 23272), and L. animalis VA 105, as possible candidates for cancer treatment and control.

Helicobacter pylori are known as a causative agent of gastritis, gastric duodenum and peptic ulcer, and gastric cancer, and multiple drug use is associated with various side effects in patients. The discovery of antibacterial substances against H. pylori from Korean resource plants is an important substitute for antibiotics. 52 species of Korean resource plants were collected and extracted with 50% ethanol, and antibacterial activity against H. pylori was measured using the disk diffusion method. The toxicity of plant extracts to human gastric adenocarcinoma(AGS) cells was measured by MTT assay, and the level of IL-8 secreted when gastric epithelial cells were inoculated with H. pylori was measured. As a result of measuring the antibacterial activity of H. pylori, antibacterial activity was confirmed in 38 plant extracts. The plant species with the strongest antibacterial activity were Chrysanthemum indicum, Rheum rhabarbarum, Patrinia scabiosaefolia and Petasites japonicus. C. indicum was not cytotoxic to H. pyroli-infected AGS cells and showed anti-inflammatory effects. This study's results can be used to develop healthy, functional foods and medical materials.