Structure Determination of Heishuixiecaoline A from Valeriana fauriei and Its Content from Different Cultivated Regions by HPLC/PDA Analysis

A germacrane-type sesquiterpenoid was isolated and purified from a methanol extract of the roots of Valeriana fauriei (RVF) through open column chromatography using silica gel. This compound was verified to be heishuixiecaoline A by spectroscopic analysis. This compound was isolated for the first time from RVF.Quantitative analysis of heishuixiecaoline A from RVF cultivated from three different regions (Eumseong, Jinbu, and Jinan regions) was performed by combining high-performance liquid chromatography with a photodiode array detector. The extract of RVF cultivated in the Jinbu region showed the highest content (9.23 mg/g). In addition, a significant amount of the compound was detected in all RVF samples, which could be expected since it is a characteristic compound of RVF. The sesquiterpenoid group heishuixiecaoline A was isolated from RVF, a resource for various pharmacological substances, and quantitative analysis of RVF cultivated from three different regions was performed. As a result of these experiments, basic data on RVF that can be used in the development and application of pharmaceuticals and health functional foods in the future were obtained.

Effects of Tianeptine on Adult Rats Following Prenatal Stress

OBJECTIVE: Exposing a pregnant female to stress during the critical period of embryonic fetal brain development increases the risk of psychiatric disorders in the offspring. The objective of this study was to investigate the effect of antidepressant tianeptine on prenatally stressed (PNS) rats. METHODS: In this study, a repeated variable stress paradigm was applied to pregnant rats during the last week of gestation. To investigate the effects of antidepressant tianeptine on PNS rats, behavioral and protein expression analyses were performed. Forced swim test, open field test, and social interaction test were performed to determine changes in PNS rats compared to non-stressed offspring. Haloperidol was used as a positive control as an antipsychotic drug based on previous studies. RESULTS: Behavioral changes were restored after treatment with tianeptine or haloperidol. Western blot and immunohistochemical analyses of the prefrontal cortex revealed downregulation of several neurodevelopmental proteins in PNS rats. After treatment with tianeptine or haloperidol, their expression levels were increased. CONCLUSION: Downregulation of several proteins in PNS rats might have caused subsequent behavioral changes in PNS rats. After tianeptine or haloperidol treatment, behavioral changes in PNS rats were restored. Therefore, tianeptine might decrease incidence of prenatal stress related-psychiatric disorders such as depression and schizophrenia.

Effects of tianeptine on symptoms of fibromyalgia via BDNF signaling in a fibromyalgia animal model

Previous reports have suggested that physical and psychological stresses may trigger fibromyalgia (FM). Stress is an important risk factor in the development of depression and memory impairments. Antidepressants have been used to prevent stress-induced abnormal pain sensation. Among various antidepressants, tianeptine has been reported to be able to prevent neurodegeneration due to chronic stress and reverse decreases in hippocampal volume. To assess the possible effect of tianeptine on FM symptoms, we constructed a FM animal model induced by restraint stress with intermittent cold stress. All mice underwent nociceptive assays using electronic von Frey anesthesiometer and Hargreaves equipment. To assess the relationship between tianeptine and expression levels of brain-derived neurotrophic factor (BDNF), cAMP response element-binding protein (CREB), and phosphorylated cAMP response element-binding protein (p-CREB), western blotting and immunohistochemistry analyses were performed. In behavioral analysis, nociception tests showed that pain threshold was significantly decreased in the FM group compared to that in the control group. Western blot and immunohistochemical analyses of medial prefrontal cortex (mPFC) and hippocampus showed downregulation of BDNF and p-CREB proteins in the FM group compared to the control group. However, tianeptine recovered these changes in behavioral tests and protein level. Therefore, this FM animal model might be useful for investigating mechanisms linking BDNF-CREB pathway and pain. Our results suggest that tianeptine might potentially have therapeutic efficacy for FM.

Gintonin, an exogenous ginseng-derived LPA receptor ligand, promotes corneal wound healing

Ginseng gintonin is an exogenous ligand of lysophosphatidic acid (LPA) receptors. Accumulating evidence shows LPA helps in rapid recovery of corneal damage. The aim of this study was to evaluate the therapeutic efficacy of gintonin in a rabbit model of corneal damage. We investigated the signal transduction pathway of gintonin in human corneal epithelium (HCE) cells to elucidate the underlying molecular mechanism. We next evaluated the therapeutic effects of gintonin, using a rabbit model of corneal damage, by undertaking histochemical analysis. Treatment of gintonin to HCE cells induced transient increases of [Ca²⁺](i) in concentration-dependent and reversible manners. Gintonin-mediated mobilization of [Ca²⁺](i) was attenuated by LPA1/3 receptor antagonist Ki16425, phospholipase C inhibitor U73122, inositol 1,4,5-triphosphate receptor antagonist 2-APB, and intracellular Ca²⁺ chelator BAPTA-AM. Gintonin facilitated in vitro wound healing in a concentration-dependent manner. When applied as an eye-drop to rabbits with corneal damage, gintonin rapidly promoted recovery. Histochemical analysis showed gintonin decreased corneal apoptosis and increased corneal cell proliferation. We demonstrated that LPA receptor activation by gintonin is linked to in vitro and in vivo therapeutic effects against corneal damage. Gintonin can be applied as a clinical agent for the rapid healing of corneal damage.

An immunohistochemical study on the initial development of 155 kd neurofascin in rat sciatic nerve / 대한수의학회지

Neurofascin, one of the members of L1CAM, has been known to have some important roles during the development of nerve fibers. In order to investigate the role of neurofascin associated with the development of nerve fibers in the rat sciatic nerve, the initial development of NF155 in the paranode was studied with immuno-fluorescence and immuno-electron microscopy. The result of the present study showed NF155 was not detected in the fetal sciatic nerve and began to reveal at the postnatal day 0 (P0) and dramatically increased by time lapse until postnatal day 7 (P7). NF155 was prominently localized in the axolemma of paranode and not detected in the central region of node of Ranvier. According to the present study, NF155 is likely to have some relationships with the formation of paranode and myelin sheath.

JNK/stress-activated protein kinase associated protein 1 is required for early development of telencephalic commissures in embryonic brains

We previously reported that mice lacking JSAP1 (jsap1-/-) were lethal and the brain of jsap1-/- at E18.5 exhibited multiple types of developmental defects, which included impaired axon projection of the corpus callosum and anterior commissures. In the current study, we examined whether the early telencephalic commissures were formed abnormally from the beginning of initial development or whether they arose normally, but have been progressively lost their maintenance in the absence of JSAP1. The early corpus callosum in the brain of jsap1+/+ at E15.5-E16.5 was found to cross the midline with forming a distinct U-shaped tract, whereas the early axonal tract in jsap1-/- appeared to cross the midline in a diffuse manner, but the lately arriving axons did not cross the midline. In the brain of jsap1-/- at E17.5, the axon terminals of lately arriving collaterals remained within each hemisphere, forming an early Probst's bundle-like shape. The early anterior commissure in the brain of jsap1+/+ at E14.5-E15.5 crossed the midline, whereas the anterior commissure in jsap1-/- developed, but was deviated from their normal path before approaching the midline. The axon tracts of the corpus callosum and anterior commissure in the brain of jsap1-/- at E16.5-E17.5 expressed phosphorylated forms of FAK and JNK, however, their expression levels in the axonal tracts were reduced compared to the respective controls in jsap1+/+. Considering the known scaffolding function of JSAP1 for the FAK and JNK pathways, these results suggest that JSAP1 is required for the pathfinding of the developing telencephalic commissures in the early brains.

Anti-inflammatory effects of 8-hydroxydeoxyguanosine in LPS-induced microglia activation: suppression of STAT3-mediated intercellular adhesion molecule-1 expression

To elucidate the roles of 8-hydroxydeoxyguanosine (oh8dG), the nucleoside of 8-hydroxyguanine (oh8Gua), we examined the effects of oh8dG upon LPS-induced intercellular adhesion molecule-1 (ICAM-1) expression and the underlying mechanisms in brain microglial cells. We found that oh8dG reduces LPS-induced reactive oxygen species (ROS) production, STAT3 activation, and ICAM-1 expression. oh8dG also suppresses pro-inflammatory cytokines, such as TNF-alpha, IL-6 and IFN-gamma. Overexpression of dominant negative STAT3 completely diminshed STAT3-mediated ICAM-1 transcriptional activity. Chromatin immunoprecipitation studies revealed that oh8dG inhibited recruitment of STAT3 to the ICAM-1 promoter, followed by a decrease in ICAM-1 expression. Using mice lacking a functional Toll-like receptor 4 (TLR4), we demonstrated that, while TLR4+/+ microglia were activated by LPS, TLR4-/-microglia exhibited inactivated STAT3 in response to LPS. Evidently, LPS modulates STAT3-dependent ICAM-1 induction through TLR4-mdiated cellular responses. Oh8dG apparently plays a role in anti-inflammatory actions via suppression of ICAM-1 gene expression by blockade of the TLR4-STAT3 signal cascade in inflammation-enhanced brain microglia. Therefore, oh8dG in the cytosol probably functions as an anti-inflammatory molecule and should be considered as a candidate for development of anti-inflammatory agents.

Clinical features of the gingiva according to maxillary anterior teeth form in adult / 대한치주과학회지

It has been suggested that morphologic characteristics of the periodontium are partly related to the shape and form of the teeth. Furthermore, the severity of symptoms of periodontal disease have been proposed to differ among these various morphologic entities or "biotypes". The purpose of the present study was to examine the relationship between the form of the crowns in the maxillary anterior tooth segment and (1) a group of morphological characteristics and (2) the thickness of the gingiva. The thickness of gingiva was measured by ultrasonic device(SDM). 100 subjects devoid of symptoms of destructive periodontal disease were examined regarding, e.g., probing depth, gingival recession, width of keratinized gingiva, thickness of the keratinized gingiva. From maxillary study cast, the width(at the apical third-CW) and the length(CL) of the crowns of the 6 anterior teeth were determined. A CW/CL-ratio was calculated for each tooth and averaged for each tooth region. The individual mean CW/CL-ratio values for the central incisors were ranked. The 10 subjects ranked highest and the 10 ranked lowest were selected as having either a long-narrow(group N) or a short-wide(group W) form of the crown of the tooth. The data for each of the examined parameters were averaged for each tooth region in each subject and mean values for subjects in groups W and N were compared using the Student t-test. Stepwise multiple regression analysis, including data from the whole sample, was performed for each tooth region with the thickness of the free gingiva as the dependent variable. The results from the analyses demonstrated that individuals with a long-narrow form of the central incisors displayed, compared to individuals with a short-wide crown, form (1) a narrow zone of keratinized gingiva, (2) a pronounced "scalloped" contour of the gingival margin. There was no significant difference between groups N and W with respect to the thickness of the keratinized gingiva. The CW/CL-ratio data revealed that a certain form of the crowns in the central incisors was accompanied by a similar form in the lateral incisors and canine tooth region. The regression analyses demonstrated that the thickness of the keratinized gingiva in central, lateral incisors and canines was significantly related to the width of the keratinized gingiva.

Clinical evaluation of full mouth disinfection therapy / 대한치주과학회지

The aim of this study is to determine whether full-mouth disinfection therapy(FMT) in our clinical setting would show better improvement of clinical parameters than partial mouth disinfection therapy(PMT) in chronic periodontitis and aggressive periodontitis patients. Among 12 patients, 6 were treated FMT and other 6 were treated PMT. Clinical parameters were calculated 3 months and 6 months after initial therapy. 1. There were no statistically significant differences between FMT and PMT in the reduction rate of bleeding on probing after 3 months, 6 months 2. Initial probing depth was 4-6mm, the mean probing depth after 3 months was 2.2mm vs 2.5mm(FMT vs PMT), after 6 months was 2.4mm vs 2.8mm. This was significantly lower in the FMT groups. 3. Initial probing depth was > or = 7mm, the reduction rate of mean probing depth during first 3 months was 4.8mm vs 4.1mm(FMT vs PMT), and 3 to 6 months was 0.5mm vs 0.3mm. This was significantly larger in the FMT groups. 4. Initial probing depth was 4-6mm, the mean clinical attachment level after 3 months was 2.3mm vs 2.7mm(FMT vs PMT), after 6 months was 2.7mm vs 3.0mm. This was significantly lower in the FMT groups. 5. Initial probing depth was > or = 7mm, the reduction rate of mean probing depth during first 3 months was 4.0mm vs 3.0mm(FMT vs PMT), and 3 to 6 months was 0mm vs -0.1mm. This was significantly larger in the FMT groups. Although the results provided us with succeccful clinical improvement in aggressive periodontitis, further research is needed to prove its additional benefit in the treatment of chronic periodontitis

Clinical evaluation of full mouth disinfection therapy / 대한치주과학회지

The aim of this study is to determine whether full-mouth disinfection therapy(FMT) in our clinical setting would show better improvement of clinical parameters than partial mouth disinfection therapy(PMT) in chronic periodontitis and aggressive periodontitis patients. Among 12 patients, 6 were treated FMT and other 6 were treated PMT. Clinical parameters were calculated 3 months and 6 months after initial therapy. 1. There were no statistically significant differences between FMT and PMT in the reduction rate of bleeding on probing after 3 months, 6 months 2. Initial probing depth was 4-6mm, the mean probing depth after 3 months was 2.2mm vs 2.5mm(FMT vs PMT), after 6 months was 2.4mm vs 2.8mm. This was significantly lower in the FMT groups. 3. Initial probing depth was > or = 7mm, the reduction rate of mean probing depth during first 3 months was 4.8mm vs 4.1mm(FMT vs PMT), and 3 to 6 months was 0.5mm vs 0.3mm. This was significantly larger in the FMT groups. 4. Initial probing depth was 4-6mm, the mean clinical attachment level after 3 months was 2.3mm vs 2.7mm(FMT vs PMT), after 6 months was 2.7mm vs 3.0mm. This was significantly lower in the FMT groups. 5. Initial probing depth was > or = 7mm, the reduction rate of mean probing depth during first 3 months was 4.0mm vs 3.0mm(FMT vs PMT), and 3 to 6 months was 0mm vs -0.1mm. This was significantly larger in the FMT groups. Although the results provided us with succeccful clinical improvement in aggressive periodontitis, further research is needed to prove its additional benefit in the treatment of chronic periodontitis

Induction of castration by immunization of male dogs with recombinant gonadotropin-releasing hormone (GnRH)-canine distemper virus (CDV) T helper cell epitope p35

Immunocastration is a considerable alternative to a surgical castration method especially in male animal species for alleviating unwanted male behaviors and characteristics. Induction of high titer of antibody specific for gonadotropin-releasing hormone (GnRH) correlates with the regression of testes. Fusion proteins composed of canine GnRH and T helper (Th) cell epitope p35 originated from canine distemper virus (CDV) F protein and goat rotavirus VP6 protein were produced in E. coli. When these fusion proteins were injected to male dogs which were previously immunized with CDV vaccine, the fusion protein of GnRH-CDV Th cell epitope p35 induced much higher antibody than that of GnRH-rotavirus VP6 protein or GnRH alone. The degeneration of spermatogenesis was also verified in the male dogs immunized with the fusion protein of GnRH-CDV Th cell epitope p35. These results indicate that canine GnRH conjugated to CDV Th cell epitope p35 acted as a strong immunogen and the antibody to GnRH specifically neutralized GnRH in the testes. This study also implies a potential application of GnRH-based vaccines for immunocastration of male pets.

Comparison of pain perception using computer-controlled anesthetic device and aspirating syringe / 대한치주과학회지

Dental phobia is the most prevalent fear in all age groups, across gender, and in all countries. One of the primary identified sources is the fear of dental injections in the dental phobia or the high dental fear and anxiety groups. The purpose of this study was to clinically evaluate the computer controlled anesthetic device and to compare it with traditional methods of dental anesthetic delivery. Fifty(mean age : 25.6 yrs) systemically and periodontally healthy volunteers participated in this study. The subjects were given contralateral buccal and palatal injections. One side was injected with the computer-controlled anesthetic device with a microprocessor and an electric motor to precisely regulate flow rate during administration : The experimental group. The control side was injected with a standard manual syringe, in which flow rate and pressure are operator-dependent and can't be controlled accurately:The control group. The subjects described their perceived pain experiences with two subjective scales. The results of this study were as follows: 1. The computer-controlled anesthetic device was significantly less painful than conventional syringe injection. 2. The female subjects reported more pain than the male subjects. But, there were no statistical differences. 3. The anesthetic effect of both methods did not show any difference. In this study, it may be concluded that pain levels decreased significantly when the computer-controlled anesthetic device was used.

The effect of third molar extraction on the periodontal status of the adjacent second molar / 대한치주과학회지

The aim of this study was to investigate the effect of third molar extraction on the periodontal status of the adjacent second molar. A total of 61 second molars in 31 adult periodontitis patients were examined. Among them, 27 second molars without adjacent third molars were included in the test group, and 34 second molars with third molar were included in the control group. Clinical parameters including plaque index, gingival index, and pocket depth and radiographic bone loss were measured around the second molar both in test and control group. The result showed that: (1) the mean plaque index and gingival index of control group were higher than these of the test group but the difference was not statistically significant, (2) the mean pocket depth of the control group was higher than the test group significantly at distal and buccal surface, (3) radiographic bone loss was greater in control group than test group significantly, (4) in Pearson correlation analysis between the age of extraction and radiographic bone loss in the test group, a positive relationship was shown(p<0.01). Within limitation of this study, it may be concluded that third molar extraction in periodontitis patients showed an improvement in periodontal status in contrast the patients group having third molar, therefore earlier a removal of third molar may minimize radiographic bone loss of the adjacent second molar.

Expression of PSD95 in the Rat Sciatic Nerve

This study was designed to elucidate the existence of PSD95 in the rat sciatic nerve. Immunohistochemical stains of cryosection and teased fiber of sciatic nerves were performed with goat polyclonal antibody against PSD95. Western blot analysis was also accomplished with the same antibody. We got an interesting result that the rat sciatic nerve obviously showed PSD95 immunoreactivity especially in the nodal and paranodal regions, and we also identified a distinct band of PSD95 by western blot. These results suggest PSD95 exists in the sciatic nerve as well as it does in the central nervous system. We suppose PSD95 may have some important roles in ion channel clustering, junctional plasticity and signal transduction in the peripheral nerves as well.

The Morphometric and Ultrastructural Study of Enteric Nervous System in Adult and Aged Rat Small Intestine / 대한해부학회지

This study was performed to investigate the morphometric and ultrastructural change in the adult and aged rat small intestine. The myenteric and submucous plexuses were stained by NADH-TR in the ileum of adult Sprague-Dawley rats (3 mo., 300~350 gm) and aged rats (24 mo., 500~550 gm). The neurons of myenteric and sumucous plexuses were divided into 3 groups depending on their cell body morphology. Type 1 cells were polygonal or round with abundant cytoplasm. Type 2 cells were spindle shaped and type 3 cells were small and round with scanty cytoplasm. The nerve cell numbers and sizes were measured using an image analyzer (VIDAS, Carl Zeiss, Co., Ltd.). Ultrastructural changes were observed by JEM-1200 EXII (JEOL Co., Ltd.) electron microscope. The result obtained are as followed: 1. In adult rats, majority of neuron population were type 3 and neuron density (total numbers/1 mm2) was more higher in submucous plexus than in the myenteric plexus. 2. Statistically significant loss of type 1 and type 2 neurons were in myenteric and submucous plexus of aged rat small intestine. 3. All types of neuron sizes were increased in aged myenteric and submucous plexuses. 4. Lipofusin granules were prominent in the cytoplasm of aged rat. Cell organelles were not shown degenerative change. These results suggest that type 1 and type 2 nerve cells which is originated from autonomic nerves were lost in aged rat small intestine. Ultrastructurally lipofusin granules were prominent in the cytoplasm of aged rat and the cell organelles were not degenerated.

The Thickness Measurement of Masticatory Mucosa on the Hard Palate and Maxillary Tuberosity in Healthy Korean Adults / 대한치주과학회지

The aim of present study was to assess the thickness of masticatory mucosa on the hard palate and tuberosity as a potential donor site for mucogingival surgery. Thickness measurement was performed in 30 dental college students who are periodontally healthy, with a recently developed, ultrasonic device(SDM). The mean age of study subjects was 23.7(range 21-29) years old and the subjects were composed of 18 males and 12 females. Eighteen standard measurement points were defined on the hard palate, located on 3 lines which ran at different distances parallel to the gingival margin. Six positions were designated on each of these 3 lines between the level of the canine and the second molar. On the tuberosity, 6 standard measurement points were defined, located on 2 lines running parallel to the gingival margin at different distances. Data were analyzed to determine differences in gender, between different positions, and between lines, by an analysis of variance. The results showed that the mucosa of the tuberosity was significantly thicker than that of the hard palate region. Gender did not influence the thickness of masticatory mucosa, either on the hard palate or the tuberosity. On the hard palate, mucosa thickness increased as the distance from the marginal gingiva increased. The mucosa over the palatal root of the maxillary first molar was significantly thinner than that at all other positions on the hard palate. Measurement error at palate was 0.25mm, at tuberosity 0.51mm. No difference in the thickness of masticatory mucosa on palate and tuberosity was found between men and women. On the hard palate, soft tissue thickness progressively increased in sites further from the gingival margin. Therefore, we may harvest more thicker graft on the tuberosity that has more masticatory mucosa thickness than hard palate, however the width may not be sufficient for using.

A study on the immunocytochemical localization of neurofascin in rat sciatic nerve

We examined the localization of neurofascin (NF) in the sciatic nerve of rat. In the myelinated fibers, neurofascin localizes strongly in the nodal axolemma except the small central cleft and also expresses in the paranodes, and weakly in the Schmidt-Lanterman incisures. In the paranodes, NF localizes around the axolemma and it expresses in the apposing membrane of paranodal loops. Axoplasm, compact myelin and cytoplasm of Schwann cell do not express NF at all. In the Schmidt-Lanterman incisures, NF is expressed weakly along the Schwann cell membrane. We propose that neurofascin may be a plasmalemmal integral protein of Schwann cell in the paranode and plays some important roles for the maintenance of axo-glial junctions at the paranode. It may also have some roles for maintaining the structure of Schmidt-Lanterman incisure and have some relations with proteins localizing in the node.