High risk human papillomavirus (HR-HPV) E2 proteins play roles in transcriptional regulation and are commonly functionally disrupted when the HPV genome integrates into host chromosomes. Some 15-40% of cancer cases, however, contain an intact E2 gene or episomal HPV. In these cases, polymorphism of the E2 gene might be involved. This study aimed to determine polymorphisms of the E2 gene in episomal HPV16 detected in high grade squamous intraepithelial lesions and squamous cell carcinomas and altered functions compared to the E2 prototype. The E2 gene was amplified and sequenced. Two expression vectors containing E2 gene polymorphisms were constructed and transfected in SiHa and C33A cells, then E6 gene as well as Il-10 and TNF-${\alpha}$ expression was determined by quantitative RT-PCR. Expression vectors and reporter vectors containing the HPV16 long control region (LCR) were co-transfected and transcriptional activity was determined. The results showed that a total of 32 nucleotides and 23 amino acids were changed in all 20 cases of study, found in the transactivation (TA) domain, hinge (H) region and DNA binding (DB) domain with 14, 5 and 13 nucleotide positions. They mostly caused amino acid change. The expressing vectors containing different E2 gene polymorphisms showed E6 mRNA suppression, TNF-${\alpha}$ mRNA suppression and IL-10 induction but no statistically significant differences when compared to the E2 prototype. Moreover, promoter activity in HPV16 LCR was not affected by E2 protein with different gene polymorphisms, in contrast to nucleotide variations in LCR that showed an effect on transcription activity. These results demonstrated that E2 gene polymorphisms of episomal HPV16 did not affect transcriptional regulation and suggested that nucleotide variation as well as epigenetic modification of the LCR might play a role in inducing malignant transformation of cells containing episomal HPV16.
This study was performed to establish an effective extraction method of pig placenta extract that could be used for a putative functional food supplement with immunomodulatory effects. In the present study, we used different temperatures (4, 37, 60, 80, and $100^{\circ}C$) and different solvents (chloroform, NaOH, and phosphate buffered saline [PBS]) to extract the pig placenta. Among the different placenta extracts yielded by the different extraction methods, placenta extract (PE) in PBS at $80^{\circ}C$ for 30 min (referred to as PE-PBS80) showed a significant increase of nitric oxide production of up to 22.97 ${\mu}M/10^5$ cells at a 1 mg/mL dose (p<0.05 ) in J774A.1 cells than other extracts and control tested. Using PE-PBS80, further animal challenges were performed to identify the immune-enhanced effects. As a result, orally administered PE-PBS80 showed a significant increase in blood T and B cell activities and immunoglobulin (IgG and IgM) production. IgG and IgM levels increased to 41.53 mg/mL at a 20 mg dose on day 7 and to 27.38 mg/mL at a 10 mg dose on day 14, respectively (p<0.05). Furthermore, PE-PBS80 was also able to significantly enhance the immune modulator cytokine levels (p<0.05) compared to the control and vehicle treatments. Among the evaluated cytokines, the tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) level increased to 28.89 pg/mL at extract doses of 20 and 50 mg, the interleukin-$1{\beta}$ (IL-$1{\beta}$) level increased to 21.52 pg/mL at extract doses of 10, 20, 50 and 75 mg and the interferon (IFN)-${\gamma}$ level increased to 18.24 pg/mL at extract doses of 10, 20, and 50 mg. Therefore, this study presents an effective method for extracting pig placenta extracts and also demonstrates that pig placenta extracts had significant immunomodulatory effects not only at the cellular level but also in a mouse model, suggesting that this material could be used as an excellent candidate functional food supplement.
Physicochemical parameters, plankton community structure, and sediment were surveyed from 1988 to 2002, at two months interval, in a eutrophic coastal lagoon (Lake Songji, Korea). The lake basin is separated from the sea by a narrow sand dune, and a shallow sill divides the lake basin into two sub-basins. The stable stratifications and chemoclines are maintained all through the year at 1-2 m depth. DO was often very low (<1 $mgO_2\;{\cdot}\;L^{-1}$) in the monimolimnion. Secchi disc transparency was in the range of 0.5-2.7 m. TP, TN, and Chl. a concentration in the mixolimnion were 0.015-0.396 $mgP\;{\cdot}\;L^{-1}$), 0.223-3.521 $mgN\;{\cdot}\;L^{-1}$, and 0.5-129.8 mg ${\cdot}\;m^{-3}$, respectively. TSI was in the eutrophic range of 54 to 62. Sediment was composed of silt and coarse silt. COD, TP, and TN content of the sediment were 51.4-116.9 $mgO_2\;{\cdot}\;gdw^{-1}$, 0.04-1.46 $mgP\;{\cdot}\;gdw^{-1}$ and, 0.12-1.03 $mgN\;{\cdot}\;gdw^{-1}$, respectively. The 49 phytoplankton species were identified. The maximum phytoplankton abundance obscured the lake in September 2001 (max. density: 23,350 cells ${\cdot}\;mL^{-1}$. The Chlorophyte Schroederia judayi was dominant species in summer (max. density: 20,417 cells ${\cdot}\;mL^{-1}$). The lake showed unique limnological features of a brackish lagoon in respect to biological community, chemical characteristics, and physical phenomena.
Jun Ha;Yongrae Kim;Cheolwoong Park;Young Choi;Jeongwoo Lee
Journal of the Korean Institute of Gas
/
v.27
no.3
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pp.52-58
/
2023
With the increasing awareness of the importance of carbon neutrality in response to global climate change, the utilization of hydrogen as a carbon-free fuel source is also growing. Hydrogen is commonly used in fuel cells (FC), but it can also be utilized in internal combustion engines (ICE) that are based on combustion. Particularly, ICEs that already have established infrastructure for production and supply can greatly contribute to the expansion of hydrogen energy utilization when it becomes difficult to rely solely on fuel cells or expand their infrastructure. However, a disadvantage of utilizing hydrogen through combustion is the potential generation of nitrogen oxides (NOx), which are harmful emissions formed when nitrogen in the air reacts with oxygen at high temperatures. In particular, for the EURO-7 exhaust regulation, which includes cold start operation, efforts to reduce exhaust emissions during the warm-up process are required. Therefore, in this study, the characteristics of nitrogen oxides and fuel consumption were investigated during the warm-up process of cooling water from room temperature to 88℃ using a 2-liter direct injection spark ignition (SI) engine fueled with hydrogen. One advantage of hydrogen, compared to conventional fuels like gasoline, natural gas, and liquefied petroleum gas (LPG), is its wide flammable range, which allows for sparser control of the excessive air ratio. In this study, the excessive air ratio was varied as 1.6/1.8/2.0 during the warm-up process, and the results were analyzed. The experimental results show that as the excessive air ratio becomes sparser during warm-up, the emission of nitrogen oxides per unit time decreases, and the thermal efficiency relatively increases. However, as the time required to reach the final temperature becomes longer, the cumulative emissions and fuel consumption may worsen.
Choi, Da Hee;Jeon, Gyeongchan;Yoon, Jihee;Min, Jiho;Park, Si Jun;Kim, Jung Su;Hwang, Ee Taek;Hwang, Hyung Seo
Journal of the Society of Cosmetic Scientists of Korea
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v.45
no.4
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pp.389-397
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2019
Lysosomes are cellular organelles involved in energy metabolism and intracellular digestion in eukaryotic cells, including protease, nuclease, glycosidase, lipase, and phosphatase. Our previous studies have confirmed that egg white lysosomes had melanin decolorization and reduction activity. However, there have been few studies on skin barrier and skin regeneration as well as inhibition of melanin production by egg white lysosomes on B16F10 melanocyte cell line. In this study, we attempted to identify the effect of lysosome-related organelle extract (LOE) extracted from egg white on the melanin content change and skin barrier enhancement in cells. First, cytotoxicity evaluation was performed on B16F10 melanocyte cell line to confirm the whitening efficacy of LOE. Cytotoxicity by LOE was not observed at 20 mg/mL concentration, but cytotoxicity was observed at 40 mg/mL, and the maximum concentration value was set to 20 mg/mL in all subsequent experiments. LOE samples of 5, 10, 20 mg/mL inhibited melanin production by 61.5 ± 4.0%, 61.4 ± 7.3%, 58.3 ± 8.3%, respectivly, compared to α-MSH, a negative control in melanin contents assay. MITF mRNA expression was reduced by about 39.7 ± 3.2% compared to the α-MSH treatment group. TEER assay using HaCaT showed that LOE increased TEER resistance in a dose-dependent manner, indicating that LOE is involved in strengthening the skin barrier. LOE also increased the TEER resistance under TNF-α treatment. Skin barrier was normally restored by LOE even under the condition of inflammation. LOE had a positive effect on cell division and cell migration promotion, confirmed by the observing the effect of promoting cell migration by LOE through cell migration assay. Taken together, we expect that LOE can be developed as a cosmetic material to enhance has effects on skin regeneration and skin barrier strengthening as well as whitening function if enzyme stabilization and formulation technology are combined.
Objectives: The aim of this study was to analyze the effects of Cheongsimyeonja-tang on the human body. Methods: This was a retrospective study, conducted using electronic medical records (EMR). Appropriate clinical cases were identified and databased through EMR. Changes of vital signs, blood tests, and functional tests before and after taking Cheongsimyeonja-tang were determined. Results: There were several significant changes after taking Cheongsimyeonja-tang. Hemoglobin, total blood proteins, and total bilirubin were increased. Eosinophil, aspartate aminotransferase, total cholesterol, and low-density lipoprotein cholesterol were decreased. Conclusions: Cheonseomyeonja-tang had significant effects, including improvement of blood cell count (increase of red blood cells and hemoglobin), improvement of blood lipid level (decrease of total cholesterol and LDL-cholesterol), and blood pressure control (reduction of systolic blood pressure).
Cervical cancer is one of the leading causes of female death. Viral oncoproteins E6 and E7 are selectively retained and expressed in carcinoma cells infected with HPV (Human pa pilloma virus) type 16 and cooperated in immotalization and transformation of primary keratinocyte. E6 and E7 oncoproteins interfere the functions of tumor suppressor proteins p53 and retinoblasoma protein (pRb), respectively. Among a lots of natural products, Mentha arvensis Linne var.piperascens have inhibitory effects on bindings between E6 oncoprotein and tumor suppressor p53, E3 ubiqutin- protein ligase (E6AP). HPV oncoprotein inhibitors from Mentha piperita L. were isolated by solvent partition and column chromatography (Silica gel, RP-18) and inhibitory compounds were finally purified by HPLC using an ELISA screening system based on binding between E6 and E6AP. The aim of this study is to identify the structure of inhibitory compounds and to investigate whether these compounds have inhibitory effects on functions of E6 oncoprotein. We investigated whether caffeic acid methyl ester (CAM) extracted from Mentha piperita L. could inhibit the function of E6 oncoprotein. CAM inhibited the in vitro binding of E6 and E6AP which are essential for the binding and degradation of the tumor suppressor p53 and also inhibited the proliferation of human cervical cancer cell lines (SiHa and CaSKi) in a dose response manner. These results suggest that CAM inhibited the function of E6 oncoprotein, suggesting that it can be used as a potential drug for the treatment of cervical cancers infected with HPV.
Background: The skin acts as a barrier to protect organisms against harmful exogenous agents. Compound K (CK) is an active metabolite of ginsenoside Rb1, Rb2 and Rc, and researchers have focused on its skin protective efficacy. In this study, we hypothesized that increased expression of the serine protease inhibitor Kazal type-5 (SPINK5) may improve skin barrier function. Methods: We screened several ginsenosides to increase SPINK5 gene promoter activity using a transactivation assay and found that CK can increase SPINK5 expression. To investigate the protective effect of CK on the skin barrier, RT-PCR and Western blotting were performed to investigate the expression levels of SPINK5, kallikrein 5 (KLK5), KLK7 and PAR2 in UVB-irradiated HaCaT cells. Measurement of transepidermal water loss (TEWL) and histological changes associated with the skin barrier were performed in a UVB-irradiated mouse model and a 1-chloro-2,4-dinitrobenzene (DNCB)-induced atopic dermatitis-like model. Results: CK treatment increased the expression of SPINK5 and decreased the expression of its downstream genes, such as KLKs and PAR2. In the UVB-irradiated mouse model and the DNCB-induced atopic dermatitis model, CK restored increased TEWL and decreased hydration and epidermal hyperplasia. In addition, CK normalized the reduced SPINK5 expression caused by UVB or DNCB, thereby restoring the expression of the proteins involved in desquamation to a level similar to normal. Conclusions: Our data showed that CK contributes to improving skin-barrier function in UVB-irradiated and DNCB-induced atopic dermatitis-like models through SPINK5. These results suggest that therapeutic attempts with CK might be useful in treating barrier-disrupted diseases.
Cervical cancer is one of the leading causes of female death. Viral oncoproteins E6 and E7 are selectively retained and expressed in carcinoma cells infected with HPV (Human papillomavirus) type 16. The HPV is cooperated in immotalization and transformation of primary keratinocyte. E6 and E7 oncoproteins interfere the functions of tumor suppressor proteins p53 and retinoblasoma protein (pRb), respectively. Among a lots of natural products, Artemisia scoparia Waldstein et Kitamura has inhibitory effects on the binding between E6 oncoprotein and tumor suppressor p53, or the binding between E6 and E6 associated protein (E6AP), an E3 ubiquitin-protein ligase. HPV oncoprotein inhibitors from Artemisia scoparia W. were isolated by solvent partition and column chromatography (Silica gel, RP-18) and the inhibitory compounds were finally purified by HPLC using an ELISA screening system based on the binding between E6 and E6AP. The aim of this study is to identify the structure of inhibitory compounds and to investigate whether these compounds have inhibitory effects on the functions of E6 oncoprotein. We investigated whether 3,5-di-O-caffeoylquinic acid (DCQA) extracted from Artemisia scoparia W. Could inhibit the function of E6 oncoprutein. DCQA inhibited the in vitro binding of E6 and E6AP which are essential for the binding and degradation of the tumor suppressor p53 and also inhibited the proliferation of human cervical cancer cell lines (SiHa and CaSKi) in a dose response manner. These results suggest that DCQA inhibited the function of E6 oncoprotein, suggesting that it can be used as a potential drug for the treatment of cervical cancers infected with HPV.
Three groups of black seabream (Acanthopagrus schlegeli) were fed with treatment diets containing certain concentrations of conjugated linoleic acid (CLA) and carotenoids. The control group feed contained 0% CLA and 0% carotenoids, the CP10 group feed contained 1% CLA and 0.1% carotenoids, and the CP25 group feed contained 2.5% CLA and 0.1% carotenoids. The CP10 and CP25 groups demonstrated the enhanced growth and increased feed conversion efficiency of black seabream. The specific growth rates (SGRs) were 0.74, 0.81, and 0.97, while the feed conversion ratios (FCRs) were 2.65, 2.46, and 2.04 for the control, CP10, and CP25 groups, respectively. The total contents of high unsaturated fatty acid (HUFA) for the control, CP10, and CP25 groups were 41.0%, 41.7%, and 43.5%, respectively. CLA was deposited to the extent of 2.8% and 5.6% in the muscle, and 4.0% and 8.3% in the viscera of the CP10 and CP25 groups, respectively. Meanwhile, treatment with the viscera lipid extract (VLE) from CP25 fish evidently lowered 3T3-L1 adipocytes viability. The lipid extract from the muscle and viscera of black seabream contained ample amounts of beneficial substances, such as CLA, carotenoids, EPA, and DHA. CLA, which enriched black seabream muscle, could be categorized as a functional food and serve as a well-being food. Meanwhile, the fish oil from its viscera could serve as a high function supplement.
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