• Maxillofacial Plastic and Reconstructive Surgery
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• v.28 no.5
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• pp.375-395
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• 2006

Purpose: Lingual nerve (LN) damage may be caused by either tumor resection or injury such as wisdom tooth extraction, Although autologous nerve graft is sometimes used to repair the damaged nerve, it has the disadvantage of necessity of another operation for nerve harvesting. Moreover, the results of nerve grafting is not satisfactory. The nerve growth factor (NGF) is well-known to play a critical role in peripheral nerve regeneration and its local delivery to the injured nerve has been continuously tried to enhance nerve regeneration. However, its application has limitations like repeated administration due to short half life of 30 minutes and an in vivo delivery model must allow for direct and local delivery. The aim of this study was to construct a well-functioning $rhNGF-{\beta}$ adenovirus for the ultimate development of improved method to promote peripheral nerve regeneration with enhanced and extended secretion of hNGF from the injured nerve by injecting $rhNGF-{\beta}$ gene directly into crush-injured LN in rat model. Materials and Methods: $hNGF-{\beta}$ gene was prepared from fetal brain cDNA library and cloned into E1/E3 deleted adenoviral vector which contains green fluorescence protein (GFP) gene as a reporter. After large scale production and purification of $rhNGF-{\beta}$ adenovirus, transfection efficiency and its expression at various cells (primary cultured Schwann cells, HEK293 cells, Schwann cell lines, NIH3T3 and CRH cells) were evaluated by fluorescent microscopy, RT-PCR, ELISA, immunocytochemistry. Furthermore, the function of rhNGF-beta, which was secreted from various cells infected with $rhNGF-{\beta}$ adenovirus, was evaluated using neuritogenesis of PC-12 cells. For in vivo evaluation of efficacy of $rhNGF-{\beta}$ adenovirus, the LNs of 8-week old rats were exposed and crush-injured with a small hemostat for 10 seconds. After the injury, $rhNGF-{\beta}$ adenovirus($2{\mu}l,\;1.5{\times}10^{11}pfu$) or saline was administered into the crushed site in the experimental (n=24) and the control group (n=24), respectively. Sham operation of another group of rats (n=9) was performed without administration of either saline or adenovirus. The taste recovery and the change of fungiform papilla were studied at 1, 2, 3 and 4 weeks. Each of the 6 animals was tested with different solutions (0.1M NaCl, 0.1M sucrose, 0.01M QHCl, or 0.01M HCl) by two-bottle test paradigm and the number of papilla was counted using SEM picture of tongue dorsum. LN was explored at the same interval as taste study and evaluated electro-physiologically (peak voltage and nerve conduction velocity) and histomorphometrically (axon count, myelin thickness). Results: The recombinant adenovirus vector carrying $rhNGF-{\beta}$ was constructed and confirmed by restriction endonuclease analysis and DNA sequence analysis. GFP expression was observed in 90% of $rhNGF-{\beta}$ adenovirus infected cells compared with uninfected cells. Total mRNA isolated from $rhNGF-{\beta}$ adenovirus infected cells showed strong RT-PCR band, however uninfected or LacZ recombinant adenovirus infected cells did not. NGF quantification by ELISA showed a maximal release of $18865.4{\pm}310.9pg/ml$ NGF at the 4th day and stably continued till 14 days by $rhNGF-{\beta}$ adenovirus infected Schwann cells. PC-12 cells exposed to media with $rhNGF-{\beta}$ adenovirus infected Schwann cell revealed at the same level of neurite-extension as the commercial NGF did. $rhNGF-{\beta}$ adenovirus injected experimental groups in comparison to the control group exhibited different taste preference ratio. Salty, sweet and sour taste preference ratio were significantly different after 2 weeks from the beginning of the experiment, which were similar to the sham group, but not to the control group.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.30 no.3 s.47
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• pp.393-397
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• 2004

L-Carnitine $({\beta}-hydroxy-{\gamma}-trimethyl-ammoniumbutyric{\;}acid)$ is a small water-soluble molecule important in mammalian fat metabolism. It is essential for the normal oxidation of fatty acids by the mitochondria, and is involved in the trans-esterification and excretion of acyl-CoA esters. In this paper, to investigate the relationship between aging and L-carnitine, we investigated the effects of in vitro matrix-metalloproteinase (MMP) inhibition and activity and expression of UYA-induced MMPs in human skin fibroblasts. Also, we studied to develop as anti-aging cosmetics with L-carnitine. Fluorometric assays of the proteolytic activities of MMP-1 (collagenase) were performed using fluorescent collagen substrates. ELISA (enzyme linked immune sorbent assay), gelatin-substrate zymography, RT-PCR ELISA techniques were used for the effects of L-carnitine on MMP expression, activity, and MMP mRNA expression in UVA irradiated fibroblast $(5\;J/cm^2)$, respectively. In addition, we performed clinical study with L-carnitine cream. L-carnitine inhibited the activities of MMP-1 in a dose-dependent manner and the $IC_{50}$ values calculated from semi-log plots were 2.45 mM, and L-carnitine showed strong inhibition on MMP-2 (gelatinase) activity in UVA irradiated fibroblast by zymography. Also, UVA induced MMP-1, 2 expression was reduced $43\%,\;53\%$ by treated with L-carnitine at 1.25 mM, and MMP-1 mRNA expression was reduced dose-dependent manner. Therefore L-carnitine was able to significantly inhibit the MMP activity, and regulate MMP expression in protein and mRNA level. The results of clinical study showed that $1.0\%$ L-carnitine treated group reduced wrinkle significantly compared with placebo treated group (P<0.05). All these results suggest that L-carnitine may be useful as new anti-aging cosmetics for protection against UVA induced Mm expression and activity.

• Journal of the Korean Society of Food Science and Nutrition
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• v.38 no.10
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• pp.1302-1309
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• 2009

This study was carried out to investigate physicochemical and functional properties of dried Brassica campestris ssp rapa (BR) sprouts. The proximate compositions of BR sprouts as dry matter basis were 2.35% of moisture content, 22.51% of crude protein, 21.60% of crude lipid, 4.35% of crude ash, and 49.19% of carbohydrate, respectively. The free sugars were identified as glucose and fructose. Analyzing total amino acids, 18 kinds of components were isolated from BR sprouts. The essential amino acid contained in BR sprouts accounted for 47.00% of total amino acid, while the non-essential amino acid accounted for 53.00%. The contents of vitamin A and vitamin E were 0.09 mg% and 3.06 mg%, respectively. Tartaric acid was the major organic acid. Among the minerals in dried BR sprouts, the content of potassium was the highest (882.50 mg%) and those of magnesium and calcium were comparatively high (342.85 mg%, 274.30 mg%). BR sprouts ethanol extract significantly inhibited the HMG-CoA reductase activity in a concentration-dependent manner in vitro. Furthermore, nitrite scavenging ability and DPPH radical scavenging activity of the ethanol extract of BR sprouts were 64.25% and 69.29% at a concentration of 1,000 ${\mu}g$/mL, respectively. These results suggest that BR sprouts possess potential antioxidative capacity and HMG-CoA reductase inhibitory activity.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• v.12 no.1
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• pp.39-45
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• 2009

Purpose: Preterm infants are very susceptible to malnutrition because of a lack of storage of nutrients, immature digestion and metabolism, and accompanying diseases associated with prematurity. The purpose of this study was to evaluate the effects of nutritional support by the pediatric nutritional support team (pNST) on the clinical course of preterm infants in the neonatal intensive care unit (NICU). Methods: Between July 2003 and July 2006, 48 preterm infants who were admitted to the NICU at Seoul National University Bundang Hospital were included. The subjects were divided into the following two subgroups according to the presence of NST activity; pre-NST group (n=23) and NST group (n=25). Hospital records were reviewed retrospectively. Results: Forty-eight preterm babies were included (M：F=27：21; gestational age, 25~33 weeks). A dietician, pharmacists, or the pNST participated in the prescription of total parenteral nutrition (TPN) more rapidly in the NST group (p=0.000). The fasting periods and TPN administration periods were significantly decreased in the NST group compared to the pre-NST group (p=0.017 & p=0.001, respectively). The doses of calories, protein, and lipids administered via TPN were significantly increased in the NST group compared to the pre-NST group (p=0.016, p=0.000, and p=0.000, respectively). The total period on antibiotic therapy was significantly decreased in the NST group compared to the pre-NST group (p=0.007). Conclusion: Because nutritional support by the pNST is of benefit to the clinical course of preterm infants in the NICU, the pNST should recommend to improve the nutritional status and clinical outcome of preterm infants.

• Childhood Kidney Diseases
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• v.9 no.2
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• pp.201-212
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• 2005

Purpose : Acute pyelonephritis is one of the most common causes of unexplained fever in children. It may lead to the development of progressive renal damage. However, the deteclion of acute pyelonephritis can be difficult, especially in infants. The objective of this study was to evaluate the diagnostic value of various lab tests and imaging studies for acute renal parenchymal changes in children with APN. We correlated the clinical and laboratory manifestations of acute pyelonephritis with the Imaging studies. Methods : We reviewed the records of 115 children (85 males and 30 females) who were hospitalized Outing the period of January 1998 to December 2002 with initial clinical symptoms suggestive of pyelonephritis. The patients' age, sex, duration of fever, laboratory findings, and causative organisms were compared with the findings of imaging studies (Technetium-99m dimercaptosuccinic acid renal scan, renal ultrasonography, intravenous pyelography, voiding cystourethrography). Results : No significant relation between the number of febrile days, leukocyte count, causative organism, and the renal abnormalities in the imaging studies were observed. On the other hand, both C-reactive protein and erythrocyte sedimentation rate levels were significantly elevated in children with positive dimercaptosuccinic acid renal scan. Furthermore, females and children older than 1 year presented with significantly higher rate of abnormal dimercaptosuccinic acid renal scan findings and vesicoureteral reflux presented by voiding cystourethrography. Conclusion : We recommend females and children older than 1 year who are suspected of acute pyelonephritis be evaluated carefully for renal involvement by performing imaging studies including dimercaptosuccinic acid renal scan and voiding cystourethrography. (J Koroan Soc Pediatr Nephrol 2005;9:201-212)

• Korean Journal of Fisheries and Aquatic Sciences
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• v.12 no.3
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• pp.191-200
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• 1979

For the use of antarctic krill as a fond protein source its compositional characteristics were investigated as the first part of the work includes other subjects such as processing of drill paste, concentrates, and fermented or seasoned product. In general composition of fresh frozen and preboiled frozen krill on board, the contents of crude fat and free amino nitrogen were higher in the former than in the latter which contained a high amount of ash. VBN was rather high as much as 37.6 and $26.4\;mg\%$ in both fresh frozen and preboiled krill. The pH of drill homogenates was 7.1 to 7.2 in both cases. Such a low pH might be attributed to a long term storage and temperature fluctuations during frequent transshipping. The amino acid competition of fresh frozen krill meat showed relatively high amount of glutamic acid, aspartic acid, lysine, proline, and leucine while methionine, histidine, serine, tyrosine, and phenylalanine were lower. Among the essential amino acids lysine and leucine were higher and methionine was lower. In tile composition of free amino acid proline, lysing, arginine, and alanine were higher comparatively to the contents of histidine, aspartic acid, serine, and threonine. It is noteworthy for nutritional qualification that tile essential amino acids particularly as lysine were abundant similarly to that of fishes. Heavy metal contents of krill meat 0.039 to 0.048 ppm as Hg, 0.06 to 0.11 ppm as Pb, less than 0.32 ppm as Zn, 0.008 to 0.012 ppm as Cd, 0.61 to 0.68 ppm as Fe, 0.87 to 1.37 ppm as Cu, and nondetective as Cr. A high Cu content seems to be resulted by tile blood pigment of crustacea. The ratio,1 of edible portion to non-edible portion were 37:63 in fresh frozen and 42:58 in preboiled frozen krill respectively. Release of drip after thawing was more in fresh frozen than in preboiled frozen drill marking $36\%$ and $24\%$ of both respectively.

• The Korean Journal of Pharmacology
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• v.30 no.3
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• pp.273-289
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• 1994

Reversible brain ischemia was produced by occluding both common carotid arteries for 5 min, and the effects of aminoguanidine (AG), $DL-{\alpha}-difluoromethylornithine$ (DFMO), MK-801, and nimodipine (NM) on the ischemia induced changes of the polyamine, glutamate and acetylcholine levels in the hippocampus CA1 subfield and the specific $[^3H]\;MK-801$ binding to the hippocampus synaptosomal membranes were studied with a histological reference of the cresyl violet stained hippocampus. The basal putrescine level $(PT:\;74.4{\pm}8.8\;nM)$ showed a rapid increase (up to 1.7 fold) for 5 min of ischemia, remained significantly increased for 6 h, and then resumed the further increase to amount gradually up to about 3 fold 96 h after recirculation. However, the level of spermidine was little changed, and the spermine level showed a transient increase during ischemia followed by a sustained decrease to about 40% of the preischemic level after recirculation. The increase of PT level induced by brain ischemia was enhanced with AG or MK-801, but it was reduced by DFMO or NM. The basal glutamate level $(GT:\;0.90{\pm}0.l4\;{\mu}M)$ rapidly increased to a peak level of $8.19{\pm}1.14\;{\mu}M$ within 5 min after onset of the ischemia and then decreased to the preischemic level in about 25 min after recirculation. And NM reduced the ischemia induced increase of GT level by about 25%, but AG, DFMO and MK-801 did not affect the GT increase. The basal acetylcholine level $(ACh:\;118.0{\pm}10.5\;{\mu}M)$ did little change during/after brain ischemia and was little affected by AG or NM. But DFMO and MK-801, respectively, produced the moderate decrease of ACh level. The specific $[^3H]\;MK-801$ binding to the hippocampus synaptosomal membrane was little affected by brain ischemia for 5 min. The control value (78.9 fmole/mg protein) was moderately decreased by AG and MK-801, respectively but was little changed by DFMO or NM. The microscopic findings of the brains extirpated on day 7 after ischemia showed severe neuronal damage of the hippocampus, particularly CA1 subfield. NM and AG moderately attenuated the delayed neuronal damage, and DFMO, on the contrary, aggravated the ischemia induced damage. However, MK-801 did not protect the hippocampus from ischemic damage. These results suggest that unlike to the mode of anti-ischemic action of NM, AG might protect the hippocampus from ischemic injury as being negatively regulatory on the N-methyl-D-aspartate (NMDA) receptor function in the hippocampus.

• Journal of the korean academy of Pediatric Dentistry
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• v.30 no.1
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• pp.171-180
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• 2003

The development of calvarial bones is tighly co-ordinated with the growth of the brain and needs of harmonious interactions between different tissues within the calvarial sutures. Premature fusion of cranial sutures, known as craniosynostosis, presumably involves disturbance of these interactions. Mutations in the homeobox-containg gene Msx2 cause human craniosynostosis syndrome. Msx genes, which are consist of Msx1, Msx2 and Msx3, are homeobox-containg transcripton factors, and were originally identified as homologue of Drosophila msh(muscle segment homeobox) gene. Msx1 and Msx2 genes, expressed mostly in overlapping patterns at multiple site of tissue interactions during vertebrate development, are associated with epithelial-mesenchymal interactions during organogenesis, targets of BMP and FGF signaling. To elucidate the function of Msx genes in the early morphogenesis of mouse cranial suture, we analyzed the expression of them by in situ hybridization during embryonic(E15-E18) stage, and did vivo experiments in E15.5 mouse using rhBMP-2, rhFGF-2 protein soaked bead. In the sagittal suture, Msx1 was expressed in the mesenchyme of suture and the dura mater, Msx2 was intensely expressed in the sutural mesenchyme and the dura mater. In the coronal suture both of Msx genes were expressed intensely in the sutural mesenchyme and expressed in the periosteum also. Msx1 had a broader expression pattern than Msx2. BMP2 beads induced expression of both Msx1 and Msx2, FGF2 beads induced expression of Msx1, but not Msx2. Taken together, these data suggest that Msx1 and Msx2 genes have important role in regulating the morphogenesis and maintenance of embryonic cranial suture. Both of Msx genes are expressed similarly but because of their upstream signaling, they function dependently or cooperatively according to change of signaling molecule.

• THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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• v.6 no.1
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• pp.81-97
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• 2000

Objectives: This experimental study was carried out to evaluate the effects of aqueous and methanol extracts of Hedyotis diffusa which has long been used for cancer treatment in oriental medicines on the induction of apoptotic cell death in human lymphoid leukemia cell line, HL-60. Methods: Cells were treated with various concentrations (200 to $0.4{\mu}g$) and periods (6 to 30 hr) of $H_2O$ and methanol extracts of Hedyotis diffusa. Then, cells were tested for viability by MTT assay. Cells wrere treated with $200{\mu}g/ml$ of methanol extract fork various periods. Genomic DNA was isolated, separated, on 1.5% agarose gels, stained with ethidium bromide and visualized under UV light. Cells were treated with $200{\mu}g/ml$ of each extract for 16 hr. Then, cells were treated with Hoechst dye 33342 and observed by fluorescence microscopy. Cells were treated with various doses of each for 12 hr and $100{\mu}g/ml$ of methanol extract for various periods. Lysate from the cells used to measure the activity of Caspase-1 and-3 proteases by using fluorogenic peptide substrates including acetyl-YVAD-AMC and acetyl-DEVD-AMC, respectively. Cells were treated with $200{\mu}g/ml$ of each extract for various periods. Cell lysates were immunoprecipated with anti-JNKl antibodies. The immune complex was reacted with $32^p-ATP$ and c-Jun as a substrate. The phosphotransferase activity of JNKI was measured by using PhosphoImage analyzer (Fuji Co., Japan). Nuclear extracts were isolated and incubated with oligonucleotide probe of $NF-{\kappa}B$. Transcriptional activation of ${\kappa}B$ was measured by using EMSA and visualized by PhosphoImage analyzer (Fuji Co, Japan). Cell lysates were prepared and analyzed by Western blotting with anti-Bc12 antibodies and anti-Bax antibodies. Cells were pretreated with various doses of methanol extract for 2 hr. Then, the extract was removed by centrifugation. Cells were resuspended with RPMI-1640 media containing 0.3% agarose, 10% FBS, overlayred onto bottom layer agarose and incubated at $CO_2$ incubator for 6 days. The number of colony was counted under light microscopy ($\time100$). Results: The death of HL-60 cells was markedly induced by the addition of methanol extract of Hedyotis diffusa in a dose and time-dependent manners. The apoptotic characteristic ladder pattern of DNA strand break was observed in death of HL-60 cells. In addition, it was shown nucleus chromatin condensation and fragmentation under Hoechst staining. Therefore, Hedyotis diffusa extract-induced death of HL-60 cells is mediated by apoptotic signaling processes. The activity of Caspase 3-like proteases remained in a basal level in HL-60 cells treated with aqueous extract of Hedyotis diffusa. However, it was markedly increased in HL-60 cells treated with methanol extract of Hedyotis diffusa. In addition, the phosphotransferase activity of JNKl was increased in HL-60 cells treated with methanol extract of Hedyotis diffusa. Furthermore, the activation of transcriptional activator, $NF-{\kappa}B$ was markedly induced by methanol extract of Hedyotis diffusa. Anti-apoptotic Bc12 was cleaved into 23Kda fragment by treatment of methanol extract of Hedyotis diffusa. However, expression of proapoptotic Bax protein was increased by treatment of methanol extract of Hedyotis diffusa in a time-dependent manner. Furthermore, methanol extract markedly inhibited the colony forming efficiency of HL-60 cells in semisolid agar culture. Conclusions: Above results suggest that methanol extract of Hedyotis diffusa induces the apoptotic death of human leukemic HL-60 cells via activations of Caspase-3 proteases, JNKI, transcriptional activator $NF-{\kappa}B$, In addition, our results also suggest that methanol extract of Hedyotis diffusa reduces the malignant potential of HL-60 cells via down regulation of colony forming effciency through cleavage of Bc12 as well as induction of Bax.

• Food Science and Preservation
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• v.23 no.1
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• pp.42-48
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• 2016

In order to investigate the utilization of the Omija (Schizandra chinensis Baillon) extract as a natural coagulant for manufacturing soybean curd, the quality characteristics of white (Baktae) and black (Seoritae) soybean curds, coagulated by the Omija extract or $MgCl_2$, were evaluated. Crude protein ($6.14{\pm}0.30$ and $6.25{\pm}0.18%$, respectively) and crude lipid ($10.86{\pm}1.74$ and $11.29{\pm}1.69%$, respectively) contents of white and black soybean curds coagulated using the Omija extract were higher than those coagulated using $MgCl_2$. Black soybean curds coagulated using the Omija extract showed higher L, a, and b values than those using $MgCl_2$. The most abundant amino acid in white and black soybean curds coagulated using the Omija extract was arginine (3.74 and 3.71 mg/100 g, dry basis, respectively). The amounts of Ca, K, Mg, and Na were the highest in both soybean curds prepared with the Omija extract. The sensory evaluation (color, flavor, taste, texture, and overall preference) showed that white and black soybean curds coagulated using the Omija extract were more preferred than those produced using $MgCl_2$. The results suggested that using the Omija extract as a natural coagulant agent could improve the quality and sensory characteristics of soybean curds.