In addition to the central and the peripheral nervous system, calcitonin gene-related peptide-like immunoreactivity (CGRP-LI) has been identified throughout the enteric nervous system. Several functions of the CGRP in gastrointestinal (G-I) tract has been identified, but the effect of CGRP on G-I motility is unclear. The distribution of calcitonin gene-related peptide-like immunoreactivity (CGRP-LI) in the murine small bowel were studied by using immunohistochemistry, also analyzed functionally by using electrophysiological method. Immunohistochemical studies demonstrated that CGRP-LI is localized in both nerve fibers and myenteric ganglion cells in the whole-mount preparation of murine small intestine. Double labelling with CGRP and c-kit investigated by confocal microscope was shown that CGRP-LI enteric nerve fiber surrounded the c-kit positive interstitial cells of Cajal (ICC). Electrophysiological finding revealed that treatment of CGRP inhibited electrical activity on culture ICC. Our results suggest a CGRP innervation of murine small bowel ICC. The released CGRP from enteric nerve terminals may induce relaxation of small bowel through the inhibition of ICC.
Animals ; Calcitonin Gene-Related Peptide* ; Calcitonin* ; Enteric Nervous System ; Ganglion Cysts ; Immunohistochemistry ; Interstitial Cells of Cajal ; Intestine, Small* ; Mice ; Nerve Fibers ; Peripheral Nervous System ; Relaxation

Interstitial cells of Cajal (ICC) are the pacemalkers in gastrointestinal muscles, and these cells also mediate or transduce inputs from the enteric nervoius system. Immunolabelling of interstitial cells of ICC in intestinal wall is recently developed by using specific marker, anti-c-kit antibody. Immunohistochemistry was done for c-Kit-positive ICC network in attempt to provide a morphological basis for the mechanism regulating gastro-intestinal movement. Cryosection and whole-mount preparations of mouse ileum and colon were immunolabelled using the anti-c-Kit. Immunolabelled specimens were observed under a confocal laser scanning microscopy. According to three dimensional reconstruction study, it was found that the c-Kit-positive cells were widely distributed in the intestinal wall: (1) circular muscle layer, (2) myenteric plexus, (3) deep muscular plexus in ileum, (4) submucosal plexus and longitudinal muscle layer in colon. The characteristic profiles of ICC containing c-Kit-positive cells provide a morphological basis upon the mechanism regulating gastro-intestinal motility.
Animals ; Colon ; Ileum ; Immunohistochemistry ; Interstitial Cells of Cajal* ; Intestines* ; Mice* ; Microscopy, Confocal ; Muscles ; Myenteric Plexus ; Submucous Plexus

OBJECTIVES:Nitric Oxide(NO) is a toxic, inorganic, gaseous free radical produced during the metabolism of L-Arginine by NO synthase(NOS). It has been implicated in a rapidly growing number of physiological and pathophysiological processes such as cytotoxic effects against microbes and tumor cells, blood vessel dilation and neurotransmitter. Recently there is growing evidence implicating NO in immune regulation, inflammation, autoimmunity, and arthritis. We performed this study to determine a role for nitric oxide in inflammatory arthritis especially rheumatoid arthritis(RA). METHODS: We measured (1) the concentrations of nitrite, a breakdown product of nitric oxide, in serum and synovial fluid from patients with RA and osteoarthritis(OA) and in the serum of controls (2) the concentrations of nitrite in the supernatant of cultured synovial tissue with RA and OA and (3) determined whether human chondrocytes and synoviocytes can synthesize nitric oxide and if so, how production is regulated by cytokines and antirheumatic drugs. RESULTS: 1) Serum nitrite concentrations in patients with RA and OA were higher than in controls. In both disease groups synovial fluid nitrite was higher than serum nitrite. Serum and synovial fluid nitrite concenrations in RA were higher than those in OA. However, those findings are not statistically significant. 2) Although these findings are not statistically significant, the concentration of nitrite in the supernatant of cultured synavial tissue with RA was higher than that in OA. 3) IL-1beta and TNF-alpah induced the biosynthesis of NO by chondrocytes and synoviocytes. IGF-1 and TGF-beta failed to provoke the production of NO. The biosynthesis of NO required an induction period of approximately 6 hours and was inhibited by L-NMMA and cycloheximide. Dexamethasone, indomethacin, gold sodium thiomalate and methotrexate had no effect on the induction of NO biosynthesis. CONCLUSION: These results suggest a role for nitric oxide as an inflommatory mediator in inflammatory arthritis.
Antirheumatic Agents ; Arginine ; Arthritis* ; Arthritis, Rheumatoid ; Autoimmunity ; Blood Cells ; Chondrocytes ; Cycloheximide ; Cytokines ; Dexamethasone ; Gold Sodium Thiomalate ; Humans ; Indomethacin ; Inflammation ; Insulin-Like Growth Factor I ; Metabolism ; Methotrexate ; Neurotransmitter Agents ; Nitric Oxide ; omega-N-Methylarginine ; Synovial Fluid ; Transforming Growth Factor beta

BACKGROUND: Patients undergoing brain surgery have a high risk of developing a number of perioperative coagulation disorders. Anesthesia and surgical stress may affect blood coagulation and fibrinolysis. The aim of this study was to evaluate perioperative changes in hemostatic parameters of patients undergoing clipping of cerebral aneurysms with a thromboelastograph (TEG) in combination with simple laboratory tests. METHODS: Twenty adult patients who had cerebral aneurysms and no history of coagulation disorders were studied. Isoflurane and N2O were used for all anesthetic proceedings. Preanesthetic, intraoperative (after skin incision and after clipping of cerebral aneurysms) and postanesthetic measurements included a TEG and simple laboratory tests. The TEG variables included r time (reaction time for clot formation), k time (clot formation time), alpha angle (rate of clot growth), MA (maximal amplitude of clot strength) and LY30 (fibrinolytic index). RESULTS: In simple laboratory tests, prothrombin time (PT) and partial thromboplastin time (PTT) at intraoperation and postanesthesia were longer than those at preanesthesia (p < 0.05). In the TEG, r and k time at intraoperation and postanesthesia were shorter than those at preanesthesia (p < 0.05). However the alpha angle at intraoperation and postanesthesia was longer than that at preanesthesia (p < 0.05). There was no significant difference in MA and LY30 except an increase in MA after the skin incision (p < 0.05) compared to the MA at preanesthesia. CONCLUSIONS: These results indicate a general hypercoagulability during and after a cerebral aneurysms operation in terms of TEG, although, the level of the PT and PTT can be at the upper limits within normal. Therefore perioperative use of coagulants in cerebral aneurysms may increase the risk of a thromboembolism because of accelerating blood coagulability. By early intraoperative and postoperativeevaluation of the hemostatic abnormality with a TEG, appropriate measures might be initiated to prevent postoperative complications due to hypercoagulability.
Adult ; Anesthesia ; Blood Coagulation* ; Brain ; Coagulants ; Fibrinolysis ; Humans ; Intracranial Aneurysm* ; Isoflurane ; Partial Thromboplastin Time ; Postoperative Complications ; Prothrombin Time ; Skin ; Thromboembolism ; Thrombophilia

OBJECTIVES: Human osteoarthritis is a heterogeneous and multifactorial disease characterized by the progressive deterioration of the cartilage of diarthrodial joints. In some instances, there is an identifiable cause, such as trauma or congenital malformation, but, mostly the etiology remains unknown. Since familial aggregation is seen, genetic factors may be important, particularly in osteoarthritis of the hand. METHODS: Blood samples from patients and controls were obtained for DNA analysis. Exon 31 of type II procollagen gene was amplified by polymerase chain reaction, and screening for the mutation was undertaken using a restriction enzyme digestion (Dsa I). RESULTS: The patients phenotype represented typical, but earlyonset and family history, osteoarthritis. No mutation in exon 31 of type II procollagen gene could be identified. CONCLUSION: Screening of the 31 exon did not, however, reveal any mutation. It needs further evaluation in other sites of type II procollagen genes.
Cartilage ; Collagen Type II ; Digestion ; DNA ; Exons ; Hand ; Humans ; Joints ; Mass Screening ; Osteoarthritis* ; Phenotype ; Polymerase Chain Reaction ; Procollagen*

Platelet-derived growth factor (PDGF) was initially described for its mitogenic activity on smooth muscle cells, fibroblast, and glial cells. The biological activities of PDGF include stimulation of mitogenesis, differentiation, wound healing, inflammation, and tumor formation. The localization of platelet-derived growth factor-alpha Receptor (PDGF-alpha R) in central nervous system was commonly restricted to oligodendrocyte progenitors during late embryonic and postnatal development. However, several studies recently demonstrated that postnatal neurons could also synthesize PDGF-alpha R in rodents. In the present study, to analyze the distributional pattern of PDGF-alpha R during postnatal development of the canine CNS, we used immunohistochemical method on sections of canine brain tissue. We found that neurons of various CNS regions, including cerebral cortex, striatum, diencephalon, nuclei of brain stem, cerebellum, spinal cord, exhibited the immunoreactivity to PDGF-alpha R as early as postnatal day 0. Generally PDGF-alpha R immunoreactivity was well localized in the dendrites and axons of neuron during the postnatal day 14 and postnatal day 28, and then showed diminished pattern. But neuronal immunoreactivity to PDGF-alpha R were maintained postnatal 6 month. These results suggest that the localization of PDGF-alpha R in postnatal developing neurons supports the several roles of PDGF for neurons including maturation and survival.
Axons ; Brain ; Brain Stem ; Central Nervous System ; Cerebellum ; Cerebral Cortex ; Dendrites ; Diencephalon ; Fibroblasts ; Immunohistochemistry ; Inflammation ; Myocytes, Smooth Muscle ; Neuroglia ; Neurons ; Oligodendroglia ; Platelet-Derived Growth Factor* ; Rodentia ; Spinal Cord ; Wound Healing

The spontaneous contractions of gastric smooth muscles are regulated by slow waves, which are modulated by both nervous system and humoral agents. This study was designed to examine the effects of Prostaglandin E2 (PGE2) on the contractile and electrical activities of antral smooth muscles in guinea-pig stomach, using an intracellular recording technique. To elucidate the underlying mechanism for its effect on contractility, ionic currents were also measured using a whole-cell patch clamp method. The basal tone by PGE2 was variable, whereas the magnitude of phasic contractions was reduced (19.0 +/- 2.1%, n=19). The resting membrane potentials were hyperpolarized (-4.4+/-0.5 mV, n=10), and plateau potentials were lowered (-2.9+/-0.5 mV, n=10). In most cases, however, the initial peak potentials of slow waves were depolarized more by PGE2 than those of control. The frequency of the slows wave was increased from 5.7+/-0.2 cycles/min to 6.5+/-0.2 (n-22). Voltage-operated Ca2+ currents were decreased by PGE2 (n=5). Voltage-operated K+ currents, both Ca-dependent and Ca-independent, were increased (n-5). These results suggest that PGE2 plays an important role in the modulation of gastric smooth muscle activities, and its inhibitory effects on the contractility and activities of slow waves are resulted from both decrease of Ca2+ currents and increase of K+ currents.
Dinoprostone* ; Membrane Potentials ; Muscle, Smooth ; Nervous System ; Stomach*

BACKGROUND: Left ventricular hypertrophy (LVH) has been known as an important predictor of prognosis of cardiovascular disease. Carboxy-terminal propeptide of procollagen type I (PIP) is related with myocardial fibrosis. We sought to analyze the differences in the characteristics of LVH, myocardial fibrosis, and LV functions among hypertension (HBP), diabetes mellitus (DM) and chronic renal failure (CRF). METHODS: We enrolled consecutive patients with LVH. Patients were grouped as HBP (n=50), DM (n=41), CRF (n=31). Age and sex-matched normal control was also enrolled (n=32). Echocardiography and blood sampling for serum PIP level measuring was performedin all participants. RESULTS: There were no differences in baseline characteristics except systolic blood pressure among four groups. In three patients groups, their LV mass indices were significantly increased than control. Serum PIP level in CRF was much higher than others (CRF 1505.5 vs. HBP 868.7 vs. DM 687.5 vs. control 826.4, p<0.0001). LV diastolic and systolic function evaluated by E', E/E, S' and midwall fractional shortening was significantly decreased in three patients groups. However, LAVi was significantly elevated and LV ejection fraction was significantly decreased in CRF compared to others. In correlation analysis, indices of diastolic function were weakly, but statistically correlated with PIP (E': r=0.234, p=0.006; LAVi: r=0.231, p=0.006). CONCLUSION: In CRF, LV function was more deteriorated and serum PIP was more elevated when compared to HBP or DM. Therefore, myocardial fibrosis may play an important role to LV dysfunction as well as LV hypertrophy in CRF in some degree.
Blood Pressure ; Cardiovascular Diseases ; Collagen Type I ; Diabetes Mellitus ; Echocardiography ; Fibrosis ; Humans ; Hypertension ; Hypertrophy ; Hypertrophy, Left Ventricular ; Kidney Failure, Chronic ; Prognosis

OBJECTIVES: Polymerase chain reaction (PCR) technology was eamine synovial fluid and peripheral T cells in patients with rheumatoid arthritis(RA) to determine the preferential usage of the T cell receptor(TCR) variable region(V) gene. METHODS: Oligonucleotide primers specific for individual TCR Vfi gene families were used to amplify the TCR gene products in a semiquantitative assay of their relative utilization in unselected T cell populations. RESULTS: The result of Vfi utilization was generally heterogenous, similar with previous reports. However, the mean expression of Vfi16 and Vfi18 in RA was more preferentially utilized compared to normal donors. The usage of Vfi in peripheral blood from 3 patients with RA demonstrated restrictions in Vfi16, Vfi 20 and Vfi18 genes, respectively. Analyses of synovial fluid resulted in restriction in Vfi12, Vfi20 and Vfi20, respectively. Although there was no significant pattern of skewed Vfi gene mean usage when comparing the synovial fluids with the peripheral blood T cells from RA patients, there were significant biased Vfi genes, Vfi12, V~I and Vfi20, each 3 patients. As the HLA type is a determining factor in shaping TCR repertoire of peripheral T cells, we compared the Vfi utilization in HLA-DR4 expressing groups that have susceptibility and gene dosage effect in disease progression. It was a little different that comparing the pattern of Vfi usage in peripheral blood and synovial fluid from RA patients between HLA-DR4 positive and negative group. CONCLUSION: The results were consistent with the conclusion that the increased Vfi family T cells infiltrate synovium and are dependent on each patient and may be involved in inducing and maintaining the synovitis that characterizes RA. The different outcome of each patient may be due to the difference in disease duration, genetic background and geographic region. A more important factor may be the stage of disease, because epitope 'induced immune reaction may change over time. Therefore, selecting patients early in the course of disease may be important and may facilitate the need for more in-depth TCR analysis in the future.
Arthritis, Rheumatoid* ; Bias (Epidemiology) ; Disease Progression ; DNA Primers ; Gene Dosage ; Genes, T-Cell Receptor ; HLA-DR4 Antigen ; Humans ; Polymerase Chain Reaction ; Synovial Fluid ; Synovial Membrane ; Synovitis ; T-Lymphocytes ; Tissue Donors