• Title/Summary/Keyword: in vitro effect

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Effects of protein content and the inclusion of protein sources with different amino acid release dynamics on the nitrogen utilization of weaned piglets

  • Hu, Nianzhi;Shen, Zhiwen;Pan, Li;Qin, Guixin;Zhao, Yuan;Bao, Nan
    • Animal Bioscience
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    • v.35 no.2
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    • pp.260-271
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    • 2022
  • Objective: We aimed to investigate the effect of the differing amino acid (AA) release dynamics of two protein sources on the growth performance, nitrogen deposition, plasma biochemical parameters, and muscle synthesis and degradation of piglets when included in their diets at normal and low concentrations. Methods: Forty-eight piglets (Duroc×Landrace×Large White) with initial body weight of 7.45±0.58 kg were assigned to six groups and fed one of 6 diets. The 6 dietary treatments were arranged by 3×2 factorial with 3 protein sources and 2 dietary protein levels. They are NCAS (a normal protein content with casein), NBlend (a normal protein content with blend of casein and corn gluten meal), NCGM (a normal protein content with corn gluten meal), LCAS (a low protein content with casein), LBlend (a low protein content with blend of casein and corn gluten meal), LCGM (a low protein content with corn gluten meal). The release dynamics of AA in these diets were determined by in vitro digestion. The digestibility, utilization and biological value of nitrogen in piglets were determined by micro Kjeldahl method. Plasma insulin was measured by enzyme-linked immunosorbent assay kits. The protein expression of mediators of muscle synthesis and degradation was determined by western blotting. Results: Although the consumption of a low-protein diet supplemented with crystalline AA was associated with greater nitrogen digestion and utilization (p<0.05), the final body weight, growth performance, nitrogen deposition, and phosphorylation of ribosomal protein S6 kinase 1 and eIF4E binding protein 1 in the muscle of pigs in the low-protein diet-fed groups were lower than those of the normal-protein diet-fed groups (p<0.05) because of the absence of non-essential AA. Because of the more balanced release of AA, the casein (CAS) and Blend-fed groups showed superior growth performance, final body weight and nitrogen deposition, and lower expression of muscle ring finger 1 and muscle atrophy F-box than the CGM-fed groups (p<0.05). Conclusion: We conclude that the balanced release of AA from CAS containing diets and mixed diets could reduce muscle degradation, favor nitrogen retention, % intake and improve growth performance in pigs consuming either a normal- or low-protein diet.

P3H4 promotes renal cell carcinoma progression and suppresses antitumor immunity via regulating GDF15-MMP9-PD-L1 axis

  • Tian, Shuo;Huang, Yan;Lai, Dong;Wang, Hanfeng;Du, Songliang;Shen, Donglai;Chen, Weihao;Xuan, Yundong;Lu, Yongliang;Feng, Huayi;Zhang, Xiangyi;Zhao, Wenlei;Wang, Chenfeng;Wang, Tao;Wu, Shengpan;Huang, Qingbo;Niu, Shaoxi;Wang, Baojun;Ma, Xin;Zhang, Xu
    • Advances in nano research
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    • v.12 no.6
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    • pp.639-652
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    • 2022
  • The prolyl 3-hydroxylase family member 4 (P3H4), is associated with post-translational modification of fibrillar collagens and aberrantly activated in cancer leading to tumor progression. However, its role in clear cell renal cell carcinoma (ccRCC) is still unknown. Here we reported that P3H4 was highly expressed in renal cancer tissues and significantly positive correlated with poor prognosis. Knockdown of P3H4 inhibited the proliferation, migration and metastasis of renal cancer cells in vitro and in vivo, and also, overexpression of it enhanced the oncogenic process. Mechanistically, P3H4 depletion decreased the levels of GDF15-MMP9 axis and repressed its downstream signaling. Further functional studies revealed that inhibition of GDF15 suppressed renal cancer cell growth and GDF15 recombinant human protein (rhGDF15) supplementation effectively rescued the inhibitory effect induced by P3H4 knockdown. Moreover, decreased levels of MMP9 caused by inhibition of P3H4-GDF15 signaling constrained the expression of PD-L1 and suppression of P3H4 accordingly promoted anti-tumor immunity via stimulating the infiltration of CD4+ and CD8+ T cells in syngeneic mice model. Taken together, our findings firstly demonstrated that P3H4 promotes ccRCC progression by activating GDF15-MMP9-PD-L1 axis and targeting P3H4-GDF15-MMP9 signaling pathway can be a novel strategy of controlling ccRCC malignancy.

The Effect of Platelet Derived Growth Factor - BB Loaded Chitosan/Calcium Metaphosphate on Bone Regeneration (혈소판유래성장인자를 함유한 Chitosan/Calcium Metaphosphate의 골조직재생효과에 관한 연구)

  • Lee, Seung-Yeol;Seol, Yang-Jo;Lee, Yong-Moo;Lee, Ju-Yeon;Lee, Seung-Jin;Kim, Suk-Young;Ku, Young;Rhyu, In-Chul;Han, Soo-Boo;Choi, Sang-Mook;Chung, Chong-Pyoung
    • Journal of Periodontal and Implant Science
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    • v.31 no.1
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    • pp.1-23
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    • 2001
  • Chitosan is biodegradable natural polymer that has been demonstrated its ability to improve wound healing, and calcium metaphosphate(CMP) is a unique class of phosphate minerals having a polymeric structure. In this study, chitosan/CMP and platelet derived growth factor(PDGF-BB) loaded chitosan/CMP sponges were developed, and the effect of the sponges on bone regeneration and their possibility as scaffolds for bone formation by three-dimensional osteoblast culture were examined. PDGF-BB loaded chitosan/CMP sponges were prepared by freeze-drying of a mixture of chitosan solution and CMP powder, and soaking in a PDGF-BB solution. Fabricated sponge retained its 3-dimensional porous structure with $100-200\;{\mu}m$ pores. The release kinetics of PDGF-BB loaded onto the sponge were measured in vitro with $^{125}I-labeled$ PDGF-BB. In order to examine their possibility as scaffolds for bone formation, fetal rat calvarial osteoblastic cells were isolated, cultured, and seeded into the sponges. The cell-sponge constructs were cultured for 28 days. Cell proliferation, alkaline phosphatase activity were measured at 1, 7, 14 and 28 days, and histologic examination was performed. In order to examine the effect on the healing of bone defect, the sponges were implanted into rat calvarial defects. Rats were sacrificed 2 and 4 weeks after implantation and histologic and histomorphometrical examination were performed. An effective therapeutic concentration of PDGF-BB following a high initial burst release was maintained throughout the examination period. PDGF-BB loaded chitosan/CMP sponges supported the proliferation of seeded osteoblastic cells as well as their differentiation as indicated by high alkaline phosphatase activities. Histologic findings indicated that seeded osteoblastic cells well attached to sponge matrices and proliferated in a multi-layer fashion. In the experiments of implantation in rat calvarial defects, histologic and histomorphometric examination revealed that chitosan/CMP sponge promoted osseous healing as compared to controls. PDGF-BB loaded chitosan/CMP sponge further echanced bone regeneration. These results suggested that PDGF-BB loaded chitosan/CMP sponge was a feasable scaffolding material to grow osteoblast in a three-dimentional structure for transplantation into a site for bone regeneration.

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Suppressive effects of ethanol extract of Aralia elata on UVB-induced oxidative stress in human keratinocytes (자외선 B를 조사한 인간유래각질세포에서 두릅순 에탄올추출물의 산화적 스트레스 억제효과)

  • Kwak, Chung Shil;Yang, Jiwon
    • Journal of Nutrition and Health
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    • v.49 no.3
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    • pp.135-143
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    • 2016
  • Purpose: Ultraviolet (UV)-induced oxidative stress contributes to several adverse biological effects on skin. Many phenolic phytochemicals have been shown to have antioxidant properties and protect skin cells from UV-induced oxidative damage. In this study, we investigated whether or not Aralia elata (AE) has a protective effect against UVB-induced reactive oxygen species (ROS), ultimately leading to photoaging. Methods: Phenolic content of dried AE and antioxidant properties of AE extract in 70% ethanol weredetermined by measuring DPPH and ABTS radical scavenging activities and ferric reducing antioxidant power (FRAP). The effect of AE extract on cellular ROS generation and expression levels of oxidative stress-response proteins such as superoxide dismutase (SOD)-1, catalase, nuclear factor-erythroid 2-related factor (Nrf)-2, and heme oxygenase (HO)-1 in UVB-irradiated ($75mJ/cm^2$) human keratinocytes (HaCaT) were further determined by 2'-7'-dichlorofluoresceine diacetate assay and Western blotting, respectively. Results: The total phenolic and flavonoid contents of dried AE were 20.15 mg tannic acid/g and 18.75 mg rutin/g, respectively. The $IC_{50}$ of AE extract against DPPH radical was $98.5{\mu}g/mL$, and ABTS radical scavenging activity and FRAP upon treatment with $1,000{\mu}g/mL$ of AE extract were $41.8{\mu}g\;ascorbic\;acid\;(AA)\;eq./mL$ and $29.7{\mu}g\;AA\;eq./mL$,m respectively. Pretreatment with AE extract significantly reduced (p < 0.05) ROS generation compared to that in UVB-irradiated control HaCaT cells. Pretreatment with AE extract reversed reduction of Nrf-2 and SOD-1 protein expression and induction of HO-1 protein expression caused by UVB exposure in HaCaT cells, whereas it did not affect catalase expression. Conclusion: AE extract in 70% ethanol demonstrated a protective effect against UVB-induced oxidative stress and decreased expression of Nrf-2 and SOD-1 in human keratinocytes. These findings suggest that AE ethanol extract might have potential as a natural resource for a skin anti-photoaging product in the food and cosmetic industry.

Antioxidative Effects of Korean Bamboo Trees, Wang-dae, Som-dae, Maengjong-juk, Jolit-dae and O-juk (한국산 왕대, 솜대, 맹종죽, 조릿대 및 오죽의 항산화 효과)

  • Lee, Min-Ja;Moon, Gap-Soon
    • Korean Journal of Food Science and Technology
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    • v.35 no.6
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    • pp.1226-1232
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    • 2003
  • Bamboo trees have been used for a long time as folk remedies for treatment of hypertension and stroke symptoms in Oriental regions. These pharmaceutical effects of bamboos look like to be related to its antioxidant capacity and phytochemicals in bamboos. To evaluate the antioxidative effects of bamboo trees, five kinds of bamboo varieties dominant in Korean peninsular were chosen and determined its total antioxidaive activities, free radical scavenging activities and nitrite scavenging activities by TEAC (Trolox Equivalent Antioxidant Capacity) assay, DPPH and Griess reagent assay using in vitro system, respectively. To evaluate the correlation between antioxidative activities and Maillard reaction during hot water extraction, contents of reducing sugar and total nitrogen and brown color intensity at 420 nm were determined. When total antioxidative activities, free radical scavenging activities and nitrite scavenging activities of five kinds of bamboo trees were compared, wang-dae (Phyllostachys bambusoides S. et Z.) showed the strongest effect among samples, although all kinds of extracts showed relatively strong effects against oxidation. The bamboo culms extract showed stronger antioxidative effects than that of bamboo leaves. In each fraction obtained from 70% ethanol extract, antioxidative effect were increased in order of dichloromethane>ethyl acetate>butanol>water>hexane fraction. In reducing sugar analysis of extracts, reducing sugar contents of water extracts were higher than that of 70% ethanol extracts and wang-dae water extract showed the highest level which was 708.92 mg/g. Total nitrogen contents of the extracts were $1.785{\sim}2.605\;mg%$ and contents in water extracts were lower than that in 70% ethanol extracts. Brown color intensity at 420 nm showed similar tendency with results in reducing sugar contents.

Inhibitory Effect of Camp Antagonist and Pka Inhibitors, and Stimulatory Effect of Adenylate Cyclase Agonist on Cathepsin K Processing in Cultured Mouse Osteoclasts (cAMP 길항제와 PKA 억제제 및 Adenylate Cyclase 촉진제의 백서 파골세포에서 Cathepsin K 생성에 대한 효과)

  • Shim, Youn-Soo
    • Journal of dental hygiene science
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    • v.6 no.1
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    • pp.1-9
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    • 2006
  • Cathepsin K (cat K) is the major cysteine protease expressed in osteoclasts and was thought to play a key role in matrix degradation during bone resorption. It was shown that the intracellular maturation of cat K was prevented by the cAMP antagonist, Rp-cAMP, and the protein kinase A (PKA) inhibitors of KT5720 and H89. In contrast, forskolin, a adenylate cyclase agonist, rather induced Cat K processing and maturation in osteoclasts. Furthermore, to determine whether cat K processing and maturation signaling involves protein kinase C (PKC), mouse total bone cells were treated with calphostin C, a specific inhibitor of PKC, however, no effect was observed, indicating that calphostin C did not affect to osteoclast-mediated cat K processing and maturation. Thus, it is indicated that the cAMP-PKA signaling pathway regulates cat K maturation in osteoclasts. Since secreted proenzymes have the potential to reenter the cell via M6P receptor, to prevent this possibility, it was tested cAMP antagonist Rp-cAMP and the PKA inhibitors KT5720 and H89 in the absence or presence of M6P. Inhibition of cat K processing by Rp-cAMP, KT5720, or H89 was observed in a dose-dependent manner. Furthermore, the addition of M6P resulted in enhanced potency of Rp-cAMP, KT5720 and H89. These dose-dependently inhibited in vitro bone resorption with a potency similar to that observed for inhibition of cat K processing.

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Inhibitory Effect of Potato Sprouting Inhibitor Chlorpropham on Dry Rot (감자 맹아억제제 Chlorpropham의 마른썩음병 억제 효과)

  • Kyusuk Han;Byung Sup Kim;Sae Jin Hong;Nam Sook Kim
    • Journal of Bio-Environment Control
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    • v.32 no.2
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    • pp.165-171
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    • 2023
  • Potato dry rot is one of the potato storage diseases caused by Fusarium species and is a representative pathological disorder that induced post-harvest loss during storage. Chlorpropham treatment for sprouting inhibition is mainly used for room temperature storage of potatoes for processing. In this study, the inhibitory effect of chlorpropham on Fusarium-induced dry rot of potato 'Dano'. To investigate the mycelial growth rate of the dry rot fungus (Fusarium solani Appel & Wollenw), mycelial growth was investigated in a chlorpropham (5.0, 50.4, 503.8, and 5,038 ppm) and prochloraz (0.1, 1.0, 10.0, and 100.0 ppm) medium containing F. oxysporum mycelia. Mycelia were more inhibited as the concentration of chlorpropham and prochloraz increased during incubation at 20℃, and the inhibition rate was 98.2% and 100% when treated with 503.8 ppm of chlorpropham and 10ppm of prochloraz in 14 days, respectively. Potato Dano tubers inoculated with F. oxysporum were dipped in chlorpropham (5.0, 50.4, and 503.8 ppm) and prochloraz (100 ppm) to investigate the effect of preventing dry rot during cold storage at 20℃ and 4℃ in vivo. The disease diameter of potatoes stored at room temperature (about 20℃) was reduced to 13.0 mm in the prochloraz 100 ppm teatment, and 10.7 mm in the chlorpropham 50.4 ppm treatment compared to 13.7 mm in the control tuber at 70 days of storage. The disease progression in all treatments including control was similar with no statistically significant difference at 4℃ air temperature. From the results of this study, it is considered that treatment with 50.4 ppm of chlorpropham after harvest will be useful for suppressing dry rot of stored potatoes.

Antioxidant Activities and Hepato-protective Effects of Stauntonia hexaphylla Fruit Extract Against H2O2-induced Oxidative Stress and Acetaminophen-induced Toxicity (멀꿀 열매 추출물의 항산화 활성 및 H2O2로 유도된 산화적 스트레스와 아세트아미노펜 독성 모델에서의 간 보호효과)

  • Lee, Gyuok;Kim, Jaeyong;Kang, Huwan;Bae, Donghyuck;Choi, Chul-yung
    • Journal of Life Science
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    • v.28 no.6
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    • pp.708-717
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    • 2018
  • The antioxidant activity and protective effects of a hot water extract from the Stauntonia hexaphylla fruit (WESHF) were investigated in vitro and in vivo. The total polyphenol and flavonoid contents of WESHF were $16.13{\pm}0.27mg$ gallic acid equivalent/g and $4.7{\pm}0.80mg$ catechin equivalent/g, respectively. In addition, the DPPH radical-scavenging activity ($SC_{50}$) and the Oxygen Radical Absorbance capacity of WESHF were $63.62{\pm}4.10{\mu}g/ml$ and $90.63{\pm}5.29{\mu}M$ trolox equivalent/g, respectively. The hepatoprotective effect of WESHF against hydrogen peroxide-induced oxidative damage was investigated. $H_2O_2$-induced liver damage on HepG2 cells was prevented by $200{\mu}g/ml$ of WESHF. Furthermore, to investigate the protection mechanism of WESHF on hydrogen peroxide-induced cytotoxicity in HepG2 cells, pre-treatment with $200{\mu}g/ml$ of WESHF significantly attenuated a decrease in the activities of CAT, SOD, GR, and GPx. The hepatoprotective activity of WESHF was evaluated in an experimental model of hepatic damage induced by acetaminophen (APAP). The levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were significantly decreased in the livers of mice treated with 200 mg/kg of WESHF compared to the APAP-treated group. The lipid peroxidation level, which increased after APAP administration, was significantly reduced in the WESHF group. In addition, histological examinations of the liver showed the same protective effect of WESHF treatment. Based on these findings, it is suggested that WESHF has potent hepatoprotective effects, and the mechanism that causes this type of protection could be related to antioxidant pathways.

Studies on the Standardization of Chinese Cabbage Kimchi (배추김치의 표준화 연구)

  • Cho, Eun-Ju;Lee, Seon-Mi;Rhee, Sook-Hee;Park, Kun-Young
    • Korean Journal of Food Science and Technology
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    • v.30 no.2
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    • pp.324-332
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    • 1998
  • In order to standardize the chinese cabbage kimchi, the preparation method, kinds of ingredients and levels of the ingredients were determined by the statistical survey of literatures obtained from cooking books, scientific papers and kimchi manufacturing factory. The standardized ingredient kinds and ratio of chinese cabbage kimchi were $13.0{\pm}7.0$ of radish, $2.0{\pm}0.5$ of green onion, $3.5{\pm}0.8\;or\;2.5{\m}0.3$ of red pepper powder, $1.4{\pm}0.4$ of garlic, $0.6{\pm}0.3$ of ginger, $2.2{\pm}1.6$ of anchovy juice, and $1.0{\pm}0.3$ of sugar in the proportion of 100 salted chinese cabbage, and the final salt concentration was adjusted to 2.7% using salt. Red pepper powder level was quite different from the literature sources, so sensory evaluation, chemical properties and antimutagenic effect and growth inhibitory effect on human cancer cells of the kimchi samples were studied to decide the proper ratio of the red pepper powder as an ingredient. Red pepper powder 3.5% (average level for kimchi manufacturing factory) added kimchi was better in quality than red pepper powder 2.5% (average level for cooking books and scientific papers) added kimchi in sensory evaluation and chemical properties. The juice of red pepper powder 3.5% added kimchi showed not only the stronger antimutagenicity against aflatoxin $B_1$ in Salmonella typhimurium TA100 but also the higher inhibitory effect on the growth of AGS human gastric adenocarcinoma cells in SRB assay than that of red pepper powder 2.5% added kimchi. In conclusion, the standardized ratio of the ingredients was 13.0 radish, 2.0 green onion, 3.5 red pepper powder, 1.4 garlic, 0.6 ginger, 2.2 anchovy juice, 1.0 sugar, and 2.7 final salt concentration in the proportion of 100 salted chinese cabbage.

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Effects of Herbal Complex on Blood Glucose in Streptozotocin-induced Diabetic Rats and in Mice Model of Metabolic Syndrome (생약복합제의 Streptozotocin 유발 당뇨 및 대사성증후군 모델 동물에서의 혈당에 미치는 효과)

  • Park, Han-Seok;Lee, Yeon-Sil;Choi, Se-Jin;Kim, Jin-Kyu;Lee, Yun-Lyul;Kim, Hyun-Gwen;Koo, Sam-Hoi;Ku, Dae-Hoy;Ki, Seung-Il;Lim, Soon-Sung
    • Korean Journal of Pharmacognosy
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    • v.40 no.3
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    • pp.196-204
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    • 2009
  • This study was carried out to investigate the in vivo and in vitro inhibitory effect of a traditional herbal complex (HC) extract prepared from a mixture of four oriental herbs (Dioscorea Rhizoma, Glycine soja Sieb. et Zucc, Bombycis corpus, Fermented Glycine soja) that have been widely used for the treatment and prevention of diabetes mellitus on hyperglycemia. The water extract of HC showed potent inhibitory effect on $\alpha$-glucosidase with $IC_{50}$ value of 1.24 mg/mL. Additionally, the ethanol extract of HC was also found to exhibit significant inhibitory effect against protein tyrosine phosphatase $1{\beta}$ ($PTP1{\beta}$), which is known as a major regulator of both insulin and leptin signaling. In the $PTP1{\beta}$ inhibitory assay, the most active n-hexane fraction obtained from the ethanol extract of HC, was identified as a mixture of fatty acid derivatives by gas chromatography-mass spectrometry (GC-MS). In high-fat diet-low dose streptozotocin (STZ)-induced diabetic rat, the water extract of HC improved the oral glucose intolerance as compared with rosiglitazone. HC also caused a marked decrease of body weight and fasting blood glucose and a significant improvement on glucose tolerance in metabolic syndrome mice model. These findings support that this traditional HC may be useful in the control of blood glucose in diabetes mellitus and metabolic syndrome.