• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.30 no.4
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• pp.37-48
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• 2017

Objectives : This Study was conducted to investigate the melanin synthesis effect of Eclipta prostrata and to determine the relationship between melanin synthesis effect of Eclipta prostrata and cAMP/PKA melanin synthesis pathway. Methods : We measured melanin contents, tyrosinase activity, expression of TRPs, p-CREB in B16F10 cells cultured with Eclipta prostrata ethanol extracts(EEP). And after treatment with H89 and dibutyryl cAMP, which inhibit or promote the activation of PKA, we observed changes in melanin synthesis and tyrosinase activity stimulated by EEP. Results : EEP increased melanin synthesis by promoting the expression of tyrosinase and TRP-1. It also promoted expression of p-CREB. H89 suppresses melanogenic effect and expression of tyrosinase, TRP-1 in B16F10 stimulated by EEP. Dibutyryl cAMP promotes melanogenic effect of EEP. Conclusions : The results of this study suggest that melanin synthesis effect of EEP is related to induction of tyrosinase and TRP-1 expression through the cAMP/PKA pathway.

• Osong Public Health and Research Perspectives
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• v.9 no.6
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• pp.325-333
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• 2018

Objectives: The purpose of this study was to determine the presence of IMP and OXA genes in clinical strains of Pseudomonas aeruginosa (P. aeruginosa) that are carriers of the ampC gene. Methods: In this study, 105 clinical isolates of P. aeruginosa were collected. Antibiotic resistance patterns were determined using the disk diffusion method. The strains carrying AmpC enzymes were characterized by a combination disk method. Multiplex-PCR was used to identify resistance and virulence genes, chi-square test was used to determine the relationship between variables. Results: Among 105 isolates of P. aeruginosa, the highest antibiotic resistance was to cefotaxime and aztreonam, and the least resistance was to colictin and ceftazidime. There were 49 isolates (46.66%) that showed an AmpC phenotype. In addition, the frequencies of the resistance genes were; OXA48 gene 85.2%, OXA199, 139 3.8%, OXA23 3.8%, OXA2 66.6%, OXA10 3.8%, OXA51 85.2% and OXA58 3.8%. The IMP27 gene was detected in 9 isolates (8.57%) and the IMP3.34 was detected in 11 isolates (10.47%). Other genes detected included; lasR (17.1%), lasB (18%) and lasA (26.6%). There was a significant relationship between virulence factors and the OX and IMP genes ($p{\leq}0.05$). Conclusion: The relationship between antibiotic resistance and virulence factors observed in this study could play an important role in outbreaks associated with P. aeruginosa infections.

• Journal of Animal Reproduction and Biotechnology
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• v.35 no.4
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• pp.357-365
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• 2020

The aim of present study was to investigate regulatory mechanism of alpha-linolenic acid (ALA) during in vitro maturation (IVM) on nuclear and cytoplasmic maturation of porcine oocytes. Basically, immature cumulus-oocyte complexes (COCs) were incubated for 22 h in IVM-I to which hormone was added, and then further incubated for 22 h in IVM-II without hormone. As a result, relative cumulus expansion was increased at 22 h after IVM and it was enhanced by treatment of ALA compared with control group (p < 0.05). During IVM process within 22 h, cAMP level in oocytes was decreased at 6 h (p < 0.05) and it was recovered at 12 h in ALA-treated group, while oocytes in control group recovered cAMP level at 22 h. In cumulus cells, it was reduced in all time point (p < 0.05) and ALA did not affect. Treatment of ALA enhanced metaphase-I (MI) and MII population of oocytes compared with oocytes in control group at 22 and 44 h, respectively (p < 0.05). Intracellular GSH levels in ALA group was increased at 22 and 44 h after IVM (p < 0.05), whereas it was increased in control group at 44 h after IVM (p < 0.05). In particular, the GSH in ALA-treated oocytes during 22 h of IVM was higher than control group at 22 h (p < 0.05). Lipid amount in oocytes from ALA group was higher than control group (p < 0.05). Treatment of ALA did not influence to absorption of glucose from medium. Cleavage and blastocyst formation of ALA-treated oocytes were enhanced compared with control group (p < 0.05). These findings suggest that supplementation of ALA could improve oocyte maturation and development competence through increasing GSH synthesis, lipid storage, and regulation of cAMP accumulation during early 22 h of IVM, and these might be mediated by cumulus expansion.

• Molecules and Cells
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• v.45 no.4
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• pp.180-192
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• 2022

Nuclear receptor coactivator 6 (NCOA6) is a transcriptional coactivator of nuclear receptors and other transcription factors. A general Ncoa6 knockout mouse was previously shown to be embryonic lethal, but we here generated liver-specific Ncoa6 knockout (Ncoa6 LKO) mice to investigate the metabolic function of NCOA6 in the liver. These Ncoa6 LKO mice exhibited similar blood glucose and insulin levels to wild type but showed improvements in glucose tolerance, insulin sensitivity, and pyruvate tolerance. The decrease in glucose production from pyruvate in these LKO mice was consistent with the abrogation of the fasting-stimulated induction of gluconeogenic genes, phosphoenolpyruvate carboxykinase 1 (Pck1) and glucose-6-phosphatase (G6pc). The forskolin-stimulated inductions of Pck1 and G6pc were also dramatically reduced in primary hepatocytes isolated from Ncoa6 LKO mice, whereas the expression levels of other gluconeogenic gene regulators, including cAMP response element binding protein (Creb), forkhead box protein O1 and peroxisome proliferator-activated receptor γ coactivator 1α, were unaltered in the LKO mouse livers. CREB phosphorylation via fasting or forskolin stimulation was normal in the livers and primary hepatocytes of the LKO mice. Notably, it was observed that CREB interacts with NCOA6. The transcriptional activity of CREB was found to be enhanced by NCOA6 in the context of Pck1 and G6pc promoters. NCOA6-dependent augmentation was abolished in cAMP response element (CRE) mutant promoters of the Pck1 and G6pc genes. Our present results suggest that NCOA6 regulates hepatic gluconeogenesis by modulating glucagon/cAMP-dependent gluconeogenic gene transcription through an interaction with CREB.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2023.04a
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• pp.56-56
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• 2023

The cone of Pinus rigida × Pinus taeda (PRT), a plant in the Pinaceae family, has long been used in traditional medicine to treat hemostasis, bruises, and burns. Previous research has shown that regulating oxidation-reduction reactions in reactive oxygen species can help inhibit melanogenesis, the process of melanin synthesis, which is a common target for addressing hyperpigmentation. Inhibiting tyrosinase is also known to be effective in this regard. Based on these findings, we conducted an investigation into the inhibitory effect of the ethyl acetate fraction of PRT (ERT) on melanogenesis in B16 F10 cells. We know that the expression levels of melanin biosynthesis-related proteins, including tyrosinase, TRP-1, and TRP-2, are regulated by MITF (microphthalmia-associated transcription factor) and cAMP, with cAMP affecting the activity of protein kinase A (PKA). PKA can reduce melanogenesis, and CREB reduces the phosphorylation of melanin-producing enzymes. In addition, the MAPK signaling pathway, composed of ERK, JNK, p38, and other factors, is also known to play a role in the inhibition of melanogenesis in melanocytes. Our immunoblotting results showed that ERT inhibited the expression of melanin production-related proteins (tyrosinase, TRP-1, TRP-2, and MITF) that were significantly increased by a-MSH treatment to promote melanin production. Furthermore, the phosphorylation levels of factors related to cAMP/PKA/CREB and MAPK signaling pathways were significantly reduced without affecting the total form. In conclusion, we believe that treatment with ERT can inhibit melanin synthesis by modulating the phosphorylation of cAMP/PKA/CREB and MAPK signaling pathways at the cellular level. These findings suggest the potential of ERT as a raw material for functional cosmetics and pharmaceuticals, thanks to its antioxidant activity and ability to inhibit melanogenesis. We thought that these findings of ERT as a natural plant resource will inspire further research and development in this area.

• Korean Journal of Organic Agriculture
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• v.31 no.4
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• pp.381-394
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• 2023

This study was conducted to monitor the antibiotics resistance of human-harmful bacteria isolated in the agricultural environment for hot peppers (Capsicum annuum) and tomato (Lycopersicon esculentum). As a result, we isolated 120 bacterial species (34 on fruits, 48 in soil, 21 in water, and 17 in manure), identified them with the 16S rRNA sequence, analyzed minimum inhibitory concentration (MIC) for 26 antibiotics using Sensititre ARIS Hi-Q system and then evaluated whether each bacterial genus acquired resistance for the tested antibiotics or not, according to the CLSI criteria. From difference in MIC between eco-friendly (EFM) and practical (PFM) cultivation farms, Klebsiella spp. isolated from EFM was resistant to ampicillin (AMP) and nalidixic acid (NAL), and that isolated from PFM was resistant to streptomycin (STR) and tetracycline (TET). Enterobacter spp. isolated from EFM was resistant to AMP and azithromycin (AZI), and that isolated from PFM was resistant to AMP, AZI, and STR. Meanwhile, Pseudomonas spp. isolated from EFM and PFM were all resistant to AMP, AZI, cefotaxime (FOT), cefoxitin (FOX), ceftriaxone (AXO), CHL, NAL, and STR. Staphylococcus spp. isolated from EFM and PFM were resistant to gentamycin (GEN), STR, and kanamycin (KAN), and in particular, that from EFM showed resistance for erythromycin (ERY). In conclusion, our study suggested that EFM lead STR antibiotics resistance for human-harmful bacteria to decrease, because only the bacteria isolated from hot pepper and tomato crop with PFM have showed resistance against STR antibiotics, regardless of bacterial genus.

• International Journal of Stem Cells
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• v.16 no.3
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• pp.315-325
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• 2023

Background and Objectives: Glioblastoma (GBM) is an aggressive primary brain tumor characterized by its heterogeneity and high recurrence and lethality rates. Glioblastoma stem cells (GSCs) play a crucial role in therapy resistance and tumor recurrence. Therefore, targeting GSCs is a key objective in developing effective treatments for GBM. The role of Parathyroid hormone-related peptide (PTHrP) in GBM and its impact on GSCs remains unclear. This study aimed to investigate the effect of PTHrP on GSCs and its potential as a therapeutic target for GBM. Methods and Results: Using the Cancer Genome Atlas (TCGA) database, we found higher expression of PTHrP in GBM, which correlated inversely with survival. GSCs were established from three human GBM samples obtained after surgical resection. Exposure to recombinant human PTHrP protein (rPTHrP) at different concentrations significantly enhanced GSCs viability. Knockdown of PTHrP using target-specific siRNA (siPTHrP) inhibited tumorsphere formation and reduced the number of BrdU-positive cells. In an orthotopic xenograft mouse model, suppression of PTHrP expression led to significant inhibition of tumor growth. The addition of rPTHrP in the growth medium counteracted the antiproliferative effect of siPTHrP. Further investigation revealed that PTHrP increased cAMP concentration and activated the PKA signaling pathway. Treatment with forskolin, an adenylyl cyclase activator, nullified the antiproliferative effect of siPTHrP. Conclusions: Our findings demonstrate that PTHrP promotes the proliferation of patient-derived GSCs by activating the cAMP/PKA signaling pathway. These results uncover a novel role for PTHrP and suggest its potential as a therapeutic target for GBM treatment.

• International Journal of Molecular Medicine
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• v.44 no.3
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• pp.1161-1171
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• 2019

The present study investigated whether glucagon like peptide-1 (GLP-1) improves glucose uptake through glucose transporter type 4 (GLUT4), mediated by the activation of sirtuin 1 (SIRT1), in skeletal muscle cells with palmitate induced-insulin resistance. The levels of glucose uptake, GLUT4, protein kinase A (PKA), and cyclic adenosine monophosphate (cAMP) were determined in human skeletal muscle myotubes (HSMMs) exposed to palmitate and GLP-1. Then, to determine whether PKA/cAMP were downstream signals of GLP-1, a PKA inhibitor was used. To determine whether SIRT-1 contributes to GLP-1 action in HSMMs with palmitate-induced insulin resistance, the levels of peroxisome proliferator-activated receptor γ coactivator 1α (PGC1α) deacetylation and SIRT-1 activity were assessed using a SIRT1 inhibitor and small interfering RNA (siRNA). The phosphorylation levels of protein kinase B (Akt) and insulin receptor substrate 1 (IRS-1) as insulin signaling pathways, were assessed in GLP-1-treated HSMMs exposed to palmitate. The influence of SIRT1 on the GLP-1-induced activation of insulin signaling pathway was determined using a SIRT1 inhibitor. GLP-1 restored the palmitate-induced reductions in the levels of glucose uptake, GLUT4 mRNA, GLUT4 promoter activity, and GLUT4 protein in HSMMs. PKA and cAMP, as GLP-1 downstream signals, played a role in this process. GLP-1 increased the deacetylation levels of PGC1α, and stimulated SIRT1 in HSMMs. Moreover, the SIRT1 inhibitor and siRNA of SIRT1 suppressed the effect of GLP-1 on GLUT4 expression in HSMMs exposed to palmitate. The SIRT1 inhibitor also prevented the GLP-1-induced phosphorylation of IRS-1 and Akt in palmitate-treated HSMMs. The present findings suggest that in palmitate-induced insulin-resistant HSMM, GLP-1 activates SIRT1 through the PKA/cAMP pathway, which in turn enhances glucose uptake through GLUT4 and the insulin signaling pathway.

• Korean Journal of Food Science and Technology
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• v.6 no.3
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• pp.177-184
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• 1974

File fish Navodon modestus was dehydrated in cabinet type hot-sir drier at $48-50^{\circ}C$ for 11 hours and also yellowfin puffer Fugu xanthopterum was dried in open air at $26-28^{\circ}C$ for 30 hours. Nucleotides and their related compounds were collected by extraction with cold perchloric acid and their amounts were determined by anion exchange column chromatography. The contents of ADP, IMP, ATP and hypoxanthine in fresh file fish muscle were 22.9, 12.1, 4.9, and 3.2 ${\mu}mole/g,$ dry wt. respectively. AMP and inosine were 0.9 ${\mu}mole/g,$ dry wt. equally. In fresh yellowfin puffer muscle, the contents of ADP, ATP, AMP, inosine and hypoxanthine were 25.6, 2.4, 1.6, 0.3, 0.6, and 0.4 ${\mu}mole/g,$ dry wt. respectively. In the case of file fish, ADP and ATP tended to degrade rapidly during hot-air dehydration. The contents of IMP were decreased slightly while AMP and inosine were increased. And another case of yellowfin puffer, ADP also tended to degrade rapidly during sun drying while AMP, IMP, inosine and hypoxanthine were increased. Especially, in both case of file fish and yellowfin puffer, inosine was increased twenty five and thirty five times during drying respectively.

• Clinical and Experimental Reproductive Medicine
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• v.14 no.2
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• pp.149-158
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• 1987

The present investigation has been undertaken to understand the mechanism of implantation process, by demonstrating the role of ovarian steroids in the differentiation of uterine endometrium for implantation. In particular, an attempt was made to examine the activity of alkaline phosphatase (ALP) in the either luminal, stroma or endometrium tissue sites under the pseudopregnant state induced by ovarian steroid hormones. Attempt was also made to demonstrate the correlate function of ovarian steroids with the cAMP concentration and prolactin level. The higher activity of ALP in the uterine endometrium was observed on day 3. However, the higher activity of ALP in the stroma and epithelium was observed on Day 6. This study, therefore, clearly demonstrates that progesterone is consecutive effect in stroma differ entiation. The cAMP concentrations on Day 3 treated with E or P was lower than those of control. On the other hand concentration on Day 6 treated with hormones was increased than those of control. It is, therefore, concluded that the concentration of cAMP in the uterine tissue undergoing differentiation is decreased. The prolactin level of the treated groups was the lower levels than those of the control groups. It is indicated that there is no effect of ovarian steroid hormone on the prolactin synthesis in this pseudopregnant state.