• Korean Journal of Pharmacognosy
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• v.25 no.3
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• pp.238-248
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• 1994

The hypoglycemic and metabolic effects of Commelina communis L. extract were investigated in alloxan induced diabetic rats. The increased blood glucose level in the diabetic rats was significantly reduced and the loss of body weight was recovered with the treatment of the plant protein fractions($30{\sim}70%$ ammonium sulfate precipitates). Administration of the plant protein fractions elicited the significant increase of glucose-6-phosphate dehydrogenase (G-6-P DH) activity and liver weight which were decreased in the diabetic rat liver. G-6-P DH was partially purified from extract- or insulin-treated diabetics, diabetic control, and normal rat liver and studied for the biochemical properties. The $K_m$ value(9.002 mM) of diabetic rat liver enzyme was greatly higher than that (0.033 mM) of normal enzyme indicating the affinity of enzyme for the substrate was significantly reduced in the diabetic rat liver. This reduced affinity of enzyme for the substrate in the diabetic rat was recovered in the extract- or insulin-treated rat liver enzyme having 0.164 or 0.208 mM of their $K_m$ values, respectively. Although there was no significant difference in the optimum pH(6.0) and optimum temperature($37^{\circ}C$) of enzyme among the experimental groups, the dependence of their activities on pH appeared to be slightly resistant in the extract- or insulin-treated group compared to the diabetic group. In order to investigate the antigenicity of rat liver enzyme among experimental groups, enzyme-linked immunosorbent assay was carried out by using anti-G-6-P DH anti-serum. Absorbance(0.102) shown in the normal rat liver was reduced even below zero in the alloxan-diabetic rat liver, but increased again in the extract- or insulin-treated rat liver(0.096 or 0.118, respectively). The result of this study suggested that G-6-P DH may be used as a marker enzyme to diagnose and to indicate the progress of the diabetics, and the hypoglycemic effect of the extracts of Commelina communis L. was certainly associated with action or mode of G-6-P DH on the rat liver.

• Biomolecules & Therapeutics
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• v.23 no.2
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• pp.189-194
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• 2015

P450 1A2 is responsible for the metabolism of clinically important drugs and the metabolic activation of environmental chemicals. Genetic variations of P450 1A2 can influence its ability to perform these functions, and thus, this study aimed to characterize the functional significance of three P450 1A2 allelic variants containing nonsynonymous single nucleotide polymorphisms (P450 $1A2^*8$, R456H; $^*15$, P42R; $^*16$, R377Q). Variants containing these SNPs were constructed and the recombinant enzymes were expressed and purified in Escherichia coli. Only the P42R variant displayed the typical CO-binding spectrum indicating a P450 holoenzyme with an expression level of ~ 170 nmol per liter culture, but no P450 spectra were observed for the two other variants. Western blot analysis revealed that the level of expression for the P42R variant was lower than that of the wild type, however the expression of variants R456H and R377Q was not detected. Enzyme kinetic analyses indicated that the P42R mutation in P450 1A2 resulted in significant changes in catalytic activities. The P42R variant displayed an increased catalytic turnover numbers ($k_{cat}$) in both of methoxyresorufin O-demethylation and phenacetin O-deethylation. In the case of phenacetin O-deethylation analysis, the overall catalytic efficiency ($k_{cat}/K_m$) increased up to 2.5 fold with a slight increase of its $K_m$ value. This study indicated that the substitution P42R in the N-terminal proline-rich region of P450 contributed to the improvement of catalytic activity albeit the reduction of P450 structural stability or the decrease of substrate affinity. Characterization of these polymorphisms should be carefully examined in terms of the metabolism of many clinical drugs and environmental chemicals.

• Journal of Veterinary Clinics
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• v.30 no.1
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• pp.57-60
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• 2013

Ingestion of grapes or raisins has been reported to the occurrence of acute renal failure (ARF) in dogs, although the mechanism remains undetermined. The prognosis often depends on the severity or clinical course of the disease at the time of presentation and is poor if the dog becomes anuric phase. To explore the characteristics and outcome of ARF caused by grape or raisin poisoning, sequentially collected data, from 2005 to 2008, of the Veterinary Medical Teaching Hospital at the Kangwon National University for clinical evaluation were retrospectively analyzed. Of the 11 clinically affected dogs, 4 cases made a full recovery, 3 died and 4 were euthanized. All but one case (raisin ingestion) had a history of grape exposure, but the exact quantity of fruit ingested was not known. The female dogs accounted for 72.7% (8 cases). Overall, the mean age was 5.3 years (range 0.2-11.3 years), and the mean body weight was 4.1 kg (range 1.4-13 kg). The average duration of hospital stay was 7.1 days (range 2-22 days). Vomiting and anorexia was reported in all dogs. Diarrhea (4 cases), oliguria (5 cases), and anuria (4 cases) with or without isosthenuria were also reported. Five dogs of 11 had mild to moderate anemia, with a decrease in packed cell volume and hemoglobin. All dogs had elevations in serum phosphorous, creatinine, and blood urea nitrogen values, but calcium values were variable; 2 dogs with hypocalcemia, 2 dogs with hypercalcemia, and the remaining 7 cases within reference interval. Dogs (n = 8) with measured on blood gas parameters had metabolic acidosis. In addition, higher serum enzyme activities were observed; amylase in 8 (72.7%) dogs, alkaline phosphatase in 7 (63.6%) dogs, and alanine aminotransferase in 5 (45.5%) dogs. Non-survived dogs revealed lower counts of platelet and lymphocyte subpopulation, as compared to the survived dogs.

• Korean Journal of Microbiology
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• v.49 no.1
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• pp.17-23
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• 2013

Candida biofilms are organized microbial communities growing on the surfaces of host tissues or indwelling medical devices, and the biofilms show enhanced resistance against the conventional antifungal agents. The roots of Coptidis chinensis have been widely used for medicinal purposes in East Asia. The present study was aimed to assess the effect of C. chinensis aqueous extract upon preformed biofilms of 10 clinical Candida albicans isolates and the antifungal activities which contribute to inhibit the C. albicans biofilm formation. Its effect on preformed biofilms was judged using XTT [2,3-Bis-(2-Methoxy-4-Nitro-5-Sulfophenyl)-2H-Tetrazolium-5-Carboxanilide)] reduction assay, and metabolic activity of all tested strains was reduced significantly ($57.3{\pm}14.7%$) at $98{\mu}g/ml$ of the C. chinensis extract. The extract damaged the cell membrane of C. albicans which was analyzed by fluorescein diacetate and propidium iodide staining. The anticandidal activity was fungicidal, and the extract obstructed the adhesion of C. albicans biofilms to polystyrene surfaces, arrested C. albicans cells at $G_o/G_1$ as well, and reduced the growth of biofilms or budding yeasts finally. The data suggest that C. chinensis has multiple antifungal effects on target fungi resulting in preventing the formation of biofilms. Therefore, C. chinensis holds great promise for exploring antifungal agents from natural products in treating and eliminating biofilm-associated Candida infection.

• Journal of Sericultural and Entomological Science
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• v.26 no.1
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• pp.1-20
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• 1984

Effects of Foliar spray of Kinetin on the growth and metabolism of mulberry tree was investigated in this studies. The results are summarized as follows; 1. Appropriate level of Kinetin for the optimal growth of mulberry tree was 100ppm. 2. In the non-fertilizer, N, P, K, and N+P+K treated plots supplemented with Kinetin, the growth of mulberry tree was generally promoted, especially in the roots, in comparison to non-supplemented plots. The effect was notably outstanding in the N, and N+P+K treated plots. 3. The mechanism by which the growth of root is stimulated was fundamentally attributed to the hypertrophy of unit cells. 4. The chlorophyll contents of the leaves in the Kinetin supplemented plots were higher than that of the non-supplemented, especially in the N, and N+P+K treated ones. 5. Likewise, total sugar contents of Kinetin supplemented plots were higher than that of the non-supplemented. Particularly the N+P+K treated plots showed higher level of sugar contents. In other plots, there were no significant differences in the level of sugar contents. 6. The activity of GOT and GPT was higher in the Kinetin supplemented plots, particularly in the N, and N+P+K treateated. 7. The contents of ascorbic acid were increased in plots with kinetin supplement in the order of N+P+K>potassium>Nitrogen>phosphorus$\geq$non-fertilizer. 8. Difference between Kinetin treatment and non-treatment was not recognized in the contents of inorganic and organic compounds.

• Journal of Nutrition and Health
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• v.32 no.6
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• pp.644-653
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• 1999

Background : Excessive intakes of $\omega$6 polyunsaturated fatty acid (PUFA) can increase oxidative stress, which may increase insulin resistance and could be the cause of metabolic syndrome X such as diabetes mellitus. One of the ways to reduce oxidative stress is the consumption of antioxidants such as vitamin E. It is controversial that vitamin E intakes may alleviate insulin resistance. The purpose of the study was whether high vitamin E intake may influence whole body glucose disposal rate(GDR), glycogen deposites, triglyceride content, lipid peroxide levels and antioxidant enzyme activities in Sprague Dawley rats fed high $\omega$6 PUFA diest. Methods : Sprague Dawley rats were divided into three groups. The control group consumed chow diet. High and low vitamin E groups consumed 40% PUFA of total energy intakes. One kilogram of diet mixture contained 300IU of $\alpha$-tocopherol in high vitamin E group, while it had 30 IU in low vitamin E group. Diets were given for 8 weeks. After 7 were of diet consumption, indwelling catheters were inserted in carotid artery and jugular vein of all rats so that GDR could be measured in awake and unstressed state. Results : Daily PUFA intakes were lower in the control group than others. Daily vitamin E intake of high vitamin E group was about ten times higher than those of low vitamin E group and the control group(p<0.0001). $\alpha$-tocopherol content in lier was highest in the high vitamin E group. GDR of the control group was 24% higher than others, and vitamin E intakes did not affect GDR. Glycogen deposit of liver in the control group was significantly higher than others, and it was not altered by vitamin E supplementation. Muscle glycogne content showed a similar tendency as liver glycogen in different diet groups. Triglyceride deposit in muscle was not different among groups. Lipid peroxide content of liver in the high vitamin E group was lower than the low of glutathione peroxidase were lowered in low vitamin E group than others, however, those of superoxide dismutase and catalase were not different. Conclusions : High vitamin E intakes can decrease oxidative stress in rats fed high (())-6 PUFA diet, but they cannot alleviate insulin resistance. Thus, increased oxidative stress through high (())-6 PUFA diet may be minimal for influencing insulin resistance.

• The Korean Journal of Nuclear Medicine
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• v.26 no.1
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• pp.14-25
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• 1992

Thallium-201 $(^{201}T1)$ SPECT studies were performed on a normal volunteer and 12 patients with intracerebral lesions: 3 patients with gliomas, 3 patients with meningiomas, 1 patient each with metastatic tumor, brain abscess, and cerebral infarction, and 3 postirradiation patients. (2 with metastatic tumors, 1 with lymphoma). A $^{201}T1$ index, based on the ratio of $^{201}T1$ uptake in the brain lesion versus the homologous contralateral brain, was calculated and compared with tumor histology and CT/MRI findings. The SPECT $^{201}T1$ scan showed minimal uptake of tracer in a normal brain. There was substantial uptake of $^{201}T1$ in high-grade gliomas (index>1.5) with little uptake in low-grade lesions. A previously irradiated patient with recurrent astrocytoma, in whom MRI study was unable to distinguish tumor recurrence from necrosis, showed the lesions with high $^{201}T1$ indices in both hemispheric regions (2.50/1.93), suggesting tumor recurrence. Two meningiomas and a metastatic tumor showed varying degrees of $^{201}T1$ uptake (index $1.71\sim8.15$), revealing that $^{201}T1$ uptake is not exclusive to high-grade gliomas. In 2 postirradiation patients with metastatic tumors, no abnormal $^{201}T1$ uptake was found in the cerebral lesions, shortly after the initiation of radiation therapy or despite the persistence of enhancing lesions-though improved-on MR images, suggesting that $^{201}T1$ uptake may reflect the metabolic and possibly clonogenic activities of tumors and the brain $^{201}T1$ SPECT imaging might be valuable for the evaluation of tumor responsiveness to the therapy and for early detection of tumor recurrence. A patient with brain abscess on antibiotic treatment, showig increased uptake of $^{201}T1$ in the resolving lesions (index 2.87/1.52) is discussed. In a patient with cerebral infarction, there was no abnormal uptake of $^{201}T1$ in infarcted tissue. When using a threshold index of 1.5, correlation rate between $^{201}T1$ uptake and contrast enhancement of the cerebral lesions on CT/MRI was 73% (8/11). In conclusion, the brain $^{201}T1$ SPECT imaging may be useful for assessment of tumor response to the therapy and to predict low-or high-grade lesions.

• The Korean Journal of Physiology
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• v.17 no.2
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• pp.125-133
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• 1983

Red cell glycolytic intermediates, metabolites and metabolic ratios were studied. Glycolytic intermediates were measured in neutralized perchloric acid extracts of red cell suspensions after 3 hr incubation at $37^{\circ}C$ in the presence and absence of saponin. Adenosine triphosphate(ATP), adenosine diphosphate(ADP), pyruvate and lactate were measured by enzymatic procedures involving stoichiometric oxidation or reduction of a pyridine nucleotide. Glucose was determined using glucose oxidase after zinc hydroxide extraction. The redox state was calculated from the lactate dehydrogenase equilibrium. Adenosine triphosphatase activity(ATPase) was measured by determining the amount of phosphate released from ATP by washed erythrocyte membranes(ghost) during 20 min. incubation. Both total hydrolysis and the amount of hydrolysis that occured in the presence of ouabain were measured. The second measurement yields Mg-ATPase and represents nonspecific ATPase activity of the membranes. The difference between total and Mg-ATPase activity can be attributed to Na-K-ATPase. For the measurement of sodium fluxes, human erythrocytes were preincubated in $^{22}Na$ for 3 hr at $37^{\circ}C$, washed and suspended in a tracer-free medium. The amount of $^{22}Na$ transported out of cells at any time was determined by analysis of supernatant samples taken at various time after addition of the labeled cells to isotope-free medium. The cells and medium were separated and the radioactivity appearing in the medium was measured. From the total radioactivity in the suspension and the radioactivity appearing in the medium at known time, the rate constant for sodium release was computed. The results are summarized as follows: 1) ATP and ATP/ADP were found to increase at every concentration of saponin tested whereas ADP declined at every cone. of saponin. The increase in pyruvate and lactate were observed at every cone, of saponin and thus $NAD^+/NADH$ computed from pyruvate/lactate also increased. Glucose utilization was stimulated by saponin. 2) $Na^+-K^+-ATPase$ activities showed a biphasic response to saponin, first increasing in lower concentration and then decreasing in higher concentration of saponin. 3) The efflux of sodium was significantly increased by saponin in the range of 5 to 10 mg%. The stimulatory effect of saponin on the rate constants for active(ouabain-sensitive) sodium efflux was inhibited by addition of ouabain.

• Reproductive and Developmental Biology
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• v.34 no.1
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• pp.47-53
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• 2010

Glycoprotein hormones have a common $\alpha$-subunit that is involved in the signaling pathway together with G protein, adenylcyclase and cAMP induction; however, it is an unclear how this common structure is related to hormonal action. To determine the biological functions of the COOH-terminal amino acids in the $\alpha$-subunit of these glycoprotein hormones, a tethered-molecule was constructed by fusing the $NH_2$-terminus of the $\alpha$-subunit to the COOH-terminus of the $\beta$-subunit of equine chorionic gonadotropin (eCG). The following deletion mutants were created by PCR; Ile was inserted at position 96 to form ${\Delta}96$, Lys was substituted at position 95 to form ${\Delta}95$, His was inserted at position 93 to form ${\Delta}93$ and Tyr was substituted at position 87 to form ${\Delta}87$. Each mutant was transfected into CHO-K1 cells. Tethered-wt eCG, and ${\Delta}96$, ${\Delta}95$, and ${\Delta}93$ mutants were efficiently secreted into the medium but the ${\Delta}87$ mutant was not secreted. Interestingly, the RT-PCR, real-time PCR, and northern blot analyses confirmed that the RNA was transcribed in the ${\Delta}87$ mutant. However, the ${\Delta}87$ mutant protein was not detected in the medium or the intracellular fraction of the cell lysates. The LH- and FSH-like activities of the recombinant proteins were assayed in terms of cAMP production using rat LH/CG and rat FSH receptors. The metabolic clearance rate (MCR) was determined by injecting rec-eCG (2 IU) into the tail vein. The ${\Delta}95$ and ${\Delta}93$ mutants were completely inactive in both the LH- and FSH-like activity assays. The ${\Delta}96$ mutant showed slight activity in the LH-like activity assay. In comparison to the wild type, the activity of the ${\Delta}96$ mutant in the FSH-like activity assay was the highest among all the mutants. The MCR assay in which rec-eCG was injected showed a peak at 10 min in all the treatment groups, which disappeared 4 h after injection. These results imply a direct interaction between the receptor and the COOH-terminal region of the a-subunit. The data also reveal a significant difference in the mechanism by which the eCG hormone interacts with the rLH and rFSH receptors. The COOH-terminal region of the $\alpha$-subunit is very important for the secretion and functioning of this hormone.

• International Journal of Oral Biology
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• v.35 no.1
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• pp.27-33
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• 2010

Periodontal disease is a major oral disorder and comprises a group of infections that lead to inflammation of the gingiva and the destruction of periodontal tissues. $PPAR{\gamma}$ plays an important role in the regulation of several metabolic pathways and has recently been implicated in inflammatory response pathways. However, its effects on periodontal inflammation have yet to be clarified. In our current study, we evaluated the anti-inflammatory effects of $PPAR{\gamma}$ on periodontal disease. Human gingival fibroblasts (HGFs) treated with lipopolysaccharide (LPS) showed high levels of intracellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), matrix metalloproteinase-2 (MMP-2), and -9 (MMP-9). Moreover, these cells also showed upregulated activities for extracellular signal regulated kinase (ERK1/2), inducible nitric oxide synthase (iNOS) and cyclooxygnase-2. However, cells treated with Ad/$PPAR{\gamma}$ and rosiglitazone in same culture system showed reduced ICAM-1, VCAM-1, MMP-2, -9 and COX-2. Finally, the anti-inflammatory effects of $PPAR{\gamma}$ appear to be mediated via the suppression of the ERK1/2 pathway and consequent inhibition of NF-kB translocation. Our present findings thus suggest that $PPAR{\gamma}$ indeed has a pivotal role in gingival inflammation and may be a putative molecular target for future therapeutic strategies to control chronic periodontal disease.