Phasic and tonic ${\gamma}$-aminobutyric acidA ($GABA_A$) receptor-mediated inhibition critically regulate neuronal information processing. As these two inhibitory modalities have distinctive features in their receptor composition, subcellular localization of receptors, and the timing of receptor activation, it has been thought that they might exert distinct roles, if not completely separable, in the regulation of neuronal function. Inhibition should be maintained and regulated depending on changes in network activity, since maintenance of excitation-inhibition balance is essential for proper functioning of the nervous system. In the present study, we investigated how phasic and tonic inhibition are maintained and regulated by different signaling cascades. Inhibitory postsynaptic currents were measured as either electrically evoked events or spontaneous events to investigate regulation of phasic inhibition in layer 2/3 pyramidal neurons of the rat visual cortex. Tonic inhibition was assessed as changes in holding currents by the application of the $GABA_A$ receptor blocker bicuculline. Basal tone of phasic inhibition was maintained by intracellular $Ca^{2+}$ and $Ca^{2+}$/calmodulin-dependent protein kinase II (CaMKII). However, maintenance of tonic inhibition relied on protein kinase A activity. Depolarization of membrane potential (5 min of 0 mV holding) potentiated phasic inhibition via $Ca^{2+}$ and CaMKII but tonic inhibition was not affected. Thus, phasic and tonic inhibition seem to be independently maintained and regulated by different signaling cascades in the same cell. These results suggest that neuromodulatory signals might differentially regulate phasic and tonic inhibition in response to changes in brain states.
This article describes the topical delivery and localization of budesonide through the hairless mouse skin. Two poly(ethylene oxide)-block-poly($\varepsilon$-caprolactone)-block-poly(ethylene oxide) (PEO-PCL-PEO) triblock copolymers (T 222 and T 252) having different CL:EO ratios were added in the preparation of budesonide particles stabilized with poly(vinyl alcohol) (PVA) and Tween 80 under ultrasonication. For comparison, a commercial PEO-PPO-PEO triblock copolymer (F68) was studied under the same condition. To demonstrate the effects of the triblock copolymer, the particle size of budesonide emulsion, entrapment efficiency, and in vitro release were measured and compared. The budesonide particles stabilized by the triblock copolymers had a diameter of ca. 350 nm with entrapment efficiencies of 66-76%. The In vitro release profiles of all samples showed an initial burst followed by sustained release. The skin penetration and permeation of budesonide were analyzed by using a Frantz diffusion cell. T 222 and T 252 exhibited higher total permeation amounts, but lower budesonide penetration amounts, than F68. The results suggest that the partitioning of budesonide in each skin layer can be adjusted in order to avoid skin thinning and negative immune response arising from the penetration of budesonide in blood vessels.
Fibronectin fragments found in the synovial fluid of patients with osteoarthritis (OA) induce the catabolic responses in cartilage. Nuclear high-mobility group protein Box 1 (HMGB1), a damage-associated molecular pattern, is responsible for the regulation of signaling pathways related to cell death and survival in response to various stimuli. In this study, we investigated whether changes induced by 29-kDa amino-terminal fibronectin fragment (29-kDa FN-f) in HMGB1 expression influences the pathogenesis of OA via an HMGB1-modulated autophagy signaling pathway. Human articular chondrocytes were enzymatically isolated from articular cartilage. The level of mRNA was measured by quantitative real-time PCR. The expression of proteins was examined by western blot analysis, immnunofluorescence assay, and enzyme-linked immunosorbent assay. Interaction of proteins was evaluated by immunoprecipitation. The HMGB1 level was significantly lower in human OA cartilage than in normal cartilage. Although 29-kDa FN-f significantly reduced the HMGB1 expression at the mRNA and protein levels 6 h after treatment, the cytoplasmic level of HMGB1 was increased in chondrocytes treated with 29-kDa FN-f, which significantly inhibited the interaction of HMGB1 with Beclin-1, increased the interaction of Bcl-2 with Beclin-1, and decreased the levels of Beclin-1 and phosphorylated Bcl-2. In addition, the level of microtubule-associated protein 1 light chain 3-II, an autophagy marker, was down-regulated in chondrocytes treated with 29-kDa FN-f, whereas the effect was antagonized by mTOR knockdown. Furthermore, prolonged treatment with 29-kDa FN-f significantly increased the release of HMGB1 into the culture medium. These results demonstrated that 29-kDa FN-f inhibits chondrocyte autophagy by modulating the HMGB1 signaling pathway.
This experimental studies was to investigate location of labeled neurons in CNS following injection of pseudorabies virus(PRV), Bartha strain, into the uterus and Sanyinjiao(Sp6) of rats. After survival times of 4-5 days following the injection of PRV, the rats were perfused, and their brain and spinal cord were frozen sectioned($30\mu\textrm{m}$). These sections were stained by PRV immunohistochemical staining methods, and observed with light microscope. The results were as follows: 1. In the spinal cord, overlap areas of PRV labeled neurons projecting to uterus and Sp6 were observed in lamina VII, IX and X areas of cervical segments. In thoracic segments, overlap areas were observed in lamina IV, VII, X and intermediolateral n.. In lumbar segments, overlap area of PRV labeled neurons were observed in lamina I, V-VII, IX, X and intermediolateral n.. In sacral segments, overlap areas of PRY labeled neurons were observed in lamina N, V, VII, X and sacral parasympathetic n.. 2. In the brain, overlap areas of PR V labeled neurons projecting to the uterus and Sp6 were observed in lateral paragigantocellular n., rostroventrolateral reticular n., raphe obscurus n., raphe pallidus n., raphe magnus n., locus coeruleus n., Barrington's n., A5 cell group, central gray n., paraventricular hypothalamic n. and arcuate n. This results suggest that overlap areas of PRV labeled neurons of the spinal cord projecting to the uterus and Sp6 might be the first-order neurons related to the viscera-somatic sensory and sympathetic preganglionic neurons. PRV labeled neurons of the brain may be the second and third-order neurons response to the movement of smooth muscle of uterus. These PRV labeled neurons may be central autonomic center related to the integration and modulation of reflex control linked to the sensory and motor system monitoring the internal environment. These overlap areas of spinal cord and brainmay be related to autonomic centers related to regulation of uterus.
Chung, Yong-Hyun;Park, Yong;Song, Tae-Yong;Jung, Jin-Ho;Gyuseong Cho
Proceedings of the Korean Society of Medical Physics Conference
/
2002.09a
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pp.331-334
/
2002
It has been reported that maximum-likelihood position-estimation (MLPE) algorithms offer advantages of improved spatial resolution and linearity over conventional Anger algorithm in gamma cameras. The purpose of this study is to evaluate the performances of the noise models, Poisson and Gaussian, in MLPE for the localization of photons in a small gamma camera (SGC) using NaI(Tl) plate and PSPMT. The SGC consists of a single NaI(Tl) crystal, 10 cm diameter and 6 mm thick, optically coupled to a PSPMT (Hamamatsu R3292-07). The PSPMT was read out using a resistive charge divider, which multiplexes 28(X) by 28(Y) cross wire anodes into four channels. Poisson and Gaussian based MLPE methods have been implemented using experimentally measured light response functions. The system resolutions estimated by Poisson and Gaussian based MLPE were 4.3 mm and 4.0 mm, respectively. Integral uniformities were 29.7% and 30.6%, linearities were 1.5 mm and 1.0 mm and count rates were 1463 cps and 1388 cps in Poisson and Gaussian based MLPE, respectively. The results indicate that Gaussian based MLPE, which is convenient to implement, has better performances and is more robust to statistical noise than Poisson based MLPE.
Objective: This study evaluated the safety and objective response of combining $^{131}I$-labeled-metuximab (Licartin) with transarterial chemoembolization (TACE) in the treatment of unresectable hepatocellular carcinoma (HCC). Materials and Methods: In a multicenter open-label clinical trial, 341 enrolled patients with stage III/IV HCC according to TNM criteria were nonrandomly assigned to a trial group (n=167) and a control group (n=174), undergoing TACE following hepatic intra-arterial injection of licartin or TACE alone from July 2007 to July 2009. Radiopharmaceutical distribution was evaluated. The primary endpoint was overall survival; secondary endpoints included time-to-progression (TTP), toxicity and adverse events (AEs). Results: The radiobiological distribution demonstrated better localization of licartin in liver tumors than other tissues (P<0.01). The organ absorbed doses to liver and red marrow were $3.19{\pm}1.01Gy$ and $0.55{\pm}0.22Gy$, respectively. The 1-year survival rate was significantly higher [79.47% vs. 65.59%, hazard ratio (HR), 0.598, P=0.041] and TTP significantly improved ($6.82{\pm}1.28$ vs. $4.7{\pm}1.14months$, P=0.037) compared with the control group. Patients at stage III achieved more benefit of one year survival than stage IV in the trial group (86.9% vs. 53.8%, P<0.001). There were significant different toxicities in leukocytopenia, thrombocytopenia and increased total bilirubin level [P<0.001, P=0.013, P<0.01, relative risk (RR) 1.63, 1.33, 1.43], but no differences in severe AEs of upper GI hemorrhage and severe liver dysfunction between the groups (5.39% vs. 2.3%, P=0.136). Conclusions: Owing to excellent tumor-targeting, promised efficacy and favourable toxicity profile, the novel combination therapy of licartin and TACE could be applied in patients with unresectable HCC.
Environmental Sciences Bulletin of The Korean Environmental Sciences Society
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v.4
no.4
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pp.231-240
/
2000
Effects of Jasmonic Acid and Wounding on Polyphenol Oxidase Activity in Senescing Tomato Leaves The effects of jasmonic acid(JA) and wounding on polyphenol oxidase(PPO) during leaf senescence was investigated by measuring the PPO activity in detached tomato(Lycopersicon esculentum Mill.) leaves of two-week-old seedlings. The PPO activity in the detached senescing leaves increased significantly in the dark. The leaf segments responded to the application of JA with accelerated senescence, as indicated by the loss of chlorophyll and rapid increase in the PPO activity. The senescence-promoting action of JA differed in the light and dark. Wounding the detached senescing leaves by scraping surface segments or making punctures with needles considerably delayed the loss of chlorophyll and had a significant effect on the PPO activity, the amounts of which were roughly proportional to the intensity of the wounding. In the dark, the combination of wounding plus JA resulted in stable levels of chlorophyll and PPO. JA and ABA acted similarly in both unwounded and wounded leaves, however, the amount of chlorophyll and PPO in the wounded segments was always higher than in the respective controls. JA was found to eliminate the senescence-retarding action of benzyladenine. In a histochemical localization test, the PPO activity was found to be localized in the cell walls of the parenchyma tissue, thereby indicating moderate cytoplasmic reactions. In the JA-treated plants, the PPO activity was intense in the cells of the cortex and phloem parenchyma. Accordingly, based on these observations it would appear that PPO is a component of a defense response maker, whereas JA plays an integral role in the intracellular signal transduction involved in inducible defense mechanisms.
The intestinal environment plays a critical role in maintaining swine health. Many factors such as diet, microbiota, and host intestinal immune response influence the intestinal environment. Intestinal alkaline phosphatase (IAP) is an important apical brush border enzyme that is influenced by these factors. IAP dephosphorylates bacterial lipopolysaccharides (LPS), unmethylated cytosine-guanosine dinucleotides, and flagellin, reducing bacterial toxicity and consequently regulating toll-like receptors (TLRs) activation and inflammation. It also desphosphorylates extracellular nucleotides such as uridine diphosphate and adenosine triphosphate, consequently reducing inflammation, modulating, and preserving the homeostasis of the intestinal microbiota. The apical localization of IAP on the epithelial surface reveals its role on LPS (from luminal bacteria) detoxification. As the expression of IAP is reported to be downregulated in piglets at weaning, LPS from commensal and pathogenic gram-negative bacteria could increase inflammatory processes by TLR-4 activation, increasing diarrhea events during this phase. Although some studies had reported potential IAP roles to promote gut health, investigations about exogenous IAP effects or feed additives modulating IAP expression and activity yet are necessary. However, we discussed in this paper that the critical assessment reported can suggest that exogenous IAP or feed additives that could increase its expression could show beneficial effects to reduce diarrhea events during the post weaning phase. Therefore, the main goals of this review are to discuss IAP's role in intestinal inflammatory processes and present feed additives used as growth promoters that may modulate IAP expression and activity to promote gut health in piglets.
Kim, Jung-Ha;Kim, Hyun-Jin;Kim, Byong-Soo;Kang, Jee-Hae;Kim, Min-Seok;Lee, Eun-Joo;Kim, Sun-Hun
International Journal of Oral Biology
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v.41
no.2
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pp.89-96
/
2016
Tooth development shows dynamic morphological changes from the stages of cap to hard tissue formation and is strictly regulated during development. In the present study, we compared expression and localization of 3 major enamel matrix proteins in rats: amelogenin, enamel and ameloblastin. DD-PCR and RT-PCR revealed differential expression of the major proteins from the cap stage to root stage. Immunofluorescence staining results indicated that amelogenin was not detected in either inner enamel epithelium or reduced enamel epithelium, but highly immunoreactive in preameloblasts and ameloblasts; in addition, it was sporadically expressed in preodontoblasts abutting preameloblasts. Ameloblastin expression was also observed in not only differentiated ameloblasts but also osteoblasts. Immunoreactivity to ameloblastin in ameloblasts was strong in Tomes' processes. Enamelin was exclusively localized along the entire newly formed and maturing enamel. Enamelin was largely localized in near Tomes' processes and enamel rods in maturing enamel. Alendronate treatment resulted in down-regulation of amelogenin and ameloblastin at both transcription and translation levels; whereas, enamelin expression was unchanged in response to the treatment. These results suggested that amelogenin, ameloblastin and enamelin might be implicated in cell differentiation, adhesion of ameloblasts to enamel and enamel crystallization during enamel matrix formation, respectively.
Kim, Hyung-Gon;Shin, Dong-Ah;Kim, Hyoung-Ihl;Yoo, Eun-Ae;Shin, Dong-Gyu;Lee, Jung-Ok
Journal of Korean Neurosurgical Society
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v.46
no.4
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pp.333-339
/
2009
Objective : Few studies on the clinical spectrum of automated pressure-controlled discography (APCD)-defined positive discs have been reported to date. Thus, the present study was undertaken to analyze clinical parameters critical for diagnosis of discogenic pain and to correlate imaging findings with intradiscal pressures and pain responses in patients with APCD-positive discs. Methods : Twenty-three patients who showed APCD-positive discs were selected for analysis. CT discogram findings and the degrees of nuclear degeneration seen on MRI were analyzed in comparison to changes of intradiscal pressure that provoked pain responses; and clinical pain patterns and dynamic factors were evaluated in relation to pain provocation. Results : Low back pain (LBP), usually centralized, with diffuse leg pain was the most frequently reported pattern of pain in these patients. Overall, LBP was most commonly induced by sitting posture, however, standing was highly correlated with L5/S1 disc lesions (p<0.01). MRI abnormalities were statistically correlated with grading of CT discogram results (p<005); with most pain response observed in CT discogram Grades 3 and 4. Pain-provoking pressure was not statistically correlated with MRI grading. However, it was higher in Grade 3 than Grade 4. Conclusion : APCD-positive discs were demonstrated in patients reporting centralized low back pain with diffuse leg pain, aggravated by sitting and standing. MRI was helpful to assess the degree of nuclear degeneration, yet it could not guarantee exact localization of the painful discs. APCD was considered to be more useful than conventional discography for diagnosis of discogenic pain.
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