Purinergic receptors play an important role in regulating gastrointestinal (GI) motility. Interstitial cells of Cajal (ICCs) are pacemaker cells that regulate GI smooth muscle activity. We studied the functional roles of external adenosine 5′-triphosphate (ATP) on pacemaker activity in cultured ICCs from mouse small intestines by using the whole-cell patch clamp technique and intracellular Ca²⁺ ([Ca²⁺]ᵢ) imaging. External ATP dose-dependently depolarized the resting membrane and produced tonic inward pacemaker currents, and these effects were antagonized by suramin, a purinergic P2 receptor antagonist. ATP-induced effects on pacemaker currents were suppressed by an external Na⁺-free solution and inhibited by the nonselective cation channel blockers, flufenamic acid and niflumic acid. The removal of external Ca²⁺ or treatment with thapsigargin (inhibitor of Ca²⁺ uptake into endoplasmic reticulum) inhibited the ATP-induced effects on pacemaker currents. Spontaneous [Ca²⁺]ᵢ oscillations were enhanced by external ATP. These results suggest that external ATP modulates pacemaker activity by activating nonselective cation channels via external Ca²⁺ influx and [Ca²⁺]ᵢ release from the endoplasmic reticulum. Thus, it seems that activating the purinergic P2 receptor may modulate GI motility by acting on ICCs in the small intestine.
Adenosine ; Adenosine Triphosphate ; Animals ; Endoplasmic Reticulum ; Flufenamic Acid ; Interstitial Cells of Cajal ; Intestine, Small ; Membranes ; Mice ; Muscle, Smooth ; Niflumic Acid ; Pacemaker, Artificial ; Receptors, Purinergic ; Receptors, Purinergic P2 ; Suramin ; Thapsigargin

The effects of antlers have long been known in traditional Asian medicine. However, few studies have investigated the effects of antlers on immunity. In this study, we investigated whether fermented antler extract (FAE) has immunomodulatory effects on spleen cells. FAE enhanced the activity of spleen cells in a concentration dependent manner compared to antler extract. Interestingly, FAE significantly increased the production of interleukin-12, a representative cytokine of cell-mediated immunity, while it marginally increased that of tumor necrosis factor-alpha. Flow cytometry analysis demonstrated that FAE can protect spleen cells from spontaneous cell death without a significant proportional change in subsets, mainly lymphocytes. Taken together, the results of the present study showed that FAE has beneficial effects on spleen cells, a major type of immune cell, indicating that it can function as an immunomodulator without significant cytotoxicity. These data may broaden the use of FAE in basic research and clinical areas.
Animals ; Antlers* ; Asian Continental Ancestry Group ; Cell Death ; Flow Cytometry ; Humans ; Immunity, Cellular ; Immunomodulation ; Interleukin-12* ; Lymphocytes ; Spleen* ; Tumor Necrosis Factor-alpha

Asparagus cochinchinensis has been used to treat various diseases including fever, cough, kidney disease, breast cancer, inflammatory disease and brain disease, while IL-4 cytokine has been considered as key regulator on the skin homeostasis and the predisposition toward allergic skin inflammation. However, few studies have investigated its effects and IL-4 correlation on skin inflammation to date. To quantitatively evaluate the suppressive effects of ethyl acetate extracts of A. cochinchinensis (EaEAC) on phthalic anhydride (PA)-induced skin inflammation and investigate the role of IL-4 during their action mechanism, alterations in general phenotype biomarkers and luciferase-derived signals were measured in IL-4/Luc/CNS-1 transgenic (Tg) mice with PA-induced skin inflammation after treatment with EaEAC for 2 weeks. Key phenotype markers including lymph node weight, immunoglobulin E (IgE) concentration, epidermis thickness and number of infiltrated mast cells were significantly decreased in the PA+EaEAC treated group compared with the PA+Vehicle treated group. In addition, expression of IL-1β and TNF-α was also decreased in the PA+EaEAC cotreated group, compared to PA+Vehicle treated group. Furthermore, a significant decrease in the luciferase signal derived from IL-4 promoter was detected in the abdominal region, submandibular lymph node and mesenteric lymph node of the PA+EaEAC treated group, compared to PA+Vehicle treated group. Taken together, these results suggest that EaEAC treatment could successfully improve PA-induced skin inflammation of IL-4/Luc/CNS-1 Tg mice, and that IL-4 cytokine plays a key role in the therapeutic process of EaEAC.
Animals ; Biomarkers ; Brain Diseases ; Cough ; Epidermis ; Fever ; Homeostasis ; Immunoglobulin E ; Immunoglobulins ; Inflammation* ; Inflammatory Breast Neoplasms ; Interleukin-4 ; Kidney Diseases ; Luciferases ; Lymph Nodes ; Mast Cells ; Mice ; Phenotype ; Skin*

This study aimed to investigate the effect of pituitary adenylate cyclase-activating peptide (PACAP) on the pacemaker activity of interstitial cells of Cajal (ICC) in mouse colon and to identify the underlying mechanisms of PACAP action. Spontaneous pacemaker activity of colonic ICC and the effects of PACAP were studied using electrophysiological recordings. Exogenously applied PACAP induced hyperpolarization of the cell membrane and inhibited pacemaker frequency in a dose-dependent manner (from 0.1 nM to 100 nM). To investigate cyclic AMP (cAMP) involvement in the effects of PACAP on ICC, SQ-22536 (an inhibitor of adenylate cyclase) and cell-permeable 8-bromo-cAMP were used. SQ-22536 decreased the frequency of pacemaker potentials, and cell-permeable 8-bromo-cAMP increased the frequency of pacemaker potentials. The effects of SQ-22536 on pacemaker potential frequency and membrane hyperpolarization were rescued by co-treatment with glibenclamide (an ATP-sensitive K+ channel blocker). However, neither N(G)-nitro-L-arginine methyl ester (L-NAME, a competitive inhibitor of NO synthase) nor 1H-[1,2,4]oxadiazolo[4,3-alpha]quinoxalin-1-one (ODQ, an inhibitor of guanylate cyclase) had any effect on PACAP-induced activity. In conclusion, this study describes the effects of PACAP on ICC in the mouse colon. PACAP inhibited the pacemaker activity of ICC by acting through ATP-sensitive K+ channels. These results provide evidence of a physiological role for PACAP in regulating gastrointestinal (GI) motility through the modulation of ICC activity.
8-Bromo Cyclic Adenosine Monophosphate ; Animals ; Cell Membrane ; Colon* ; Cyclic AMP ; Glyburide ; Interstitial Cells of Cajal* ; Membranes ; Mice ; NG-Nitroarginine Methyl Ester ; Pituitary Adenylate Cyclase-Activating Polypeptide*

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

BACKGROUND AND OBJECTIVES: It has been suggested that nitric oxide (NO) and atrial natriuretic peptide (ANP) share a final common pathway for vascular smooth muscle relaxation. The aim of the present study was to determine the role of NO on the hypotensive and vasorelaxant effects of ANP. MATERIALS AND METHODS: Sprague-Dawley rats weighing 250-300 g each were anesthetized with thiopental (50 mg/kg IP). The femoral artery was cannulated and the arterial blood pressure and heart rate were continuously monitored in the anesthetized rats (n=19). ANP was administered into the jugular vein after L-NAME treatment. In vitro experiments were performed on intact and endothelium-denuded isolated thoracic aortic rings (n=51) in the presence of either L-NAME or methylene blue. RESULTS: Intravenous administration of ANP (5 ug/kg bolus and 0.2 ug/kg/min infusion) caused a decrease in the mean arterial pressure. L-NAME-pretreatment (1 mg/kg) suppressed the depressor response of ANP. In vitro, the ANP caused a dose-dependent relaxation, and the relaxation response to ANP was attenuated by L-NAME (10-4 M). Endothelium removal or methylene blue (10-5 M) also inhibited the ANP-induced vascular relaxation. CONCLUSION: These results suggest that the hypotensive and the vasorelaxant effect of ANP are, at least in part, NO-dependent.
Administration, Intravenous ; Animals ; Arterial Pressure ; Atrial Natriuretic Factor ; Endothelium ; Femoral Artery ; Heart Rate ; Jugular Veins ; Methylene Blue ; Muscle, Smooth, Vascular ; NG-Nitroarginine Methyl Ester ; Nitric Oxide* ; Rats* ; Rats, Sprague-Dawley ; Relaxation ; Thiopental

The indole alkaloid harmaline has been to cause tremor and ataxia, and produce cerebellar neurotoxicity in rat. Degeneration of Purkinje cell alligned in narrow parasagittal bands result from excitation of inferior olivary nucleus in harmaline-treated rats. The objective of this study was to investigate the hypothesis that excitation of climbing fiberinduced by harmaline mediates Purkinje cell injury or degeneration. For this purpose, the inferior olive of rats was chemically ablated by using 3-acetyl pyridine, a neurotoxic chemical, and cerebellar damage followed by administration of harmaline was analyzed using immunohistochemical markers for neurons, glial cells. The results demonstrated that a subset of Purkinje cell in the vermis and paravermis degenerated after harmaline treatment, but harmaline produced little or no Purkinje cell degeneration after inferior olivary ablation. These results suggested that harmalineinduced activation of inferior olivary neurons may lead to release of glutamate from climbing fiber synaptic terminal distributed over the Purkinje cells, and may lead to cytotoxic degeneration of Purkinje cells.
Animals ; Ataxia ; Cerebellum ; Glutamic Acid ; Harmaline* ; Neuroglia ; Neurons ; Olea ; Olivary Nucleus ; Presynaptic Terminals ; Purkinje Cells* ; Rats ; Tremor

BACKGROUND AND OBJECTIVES: The vascular endothelium plays an important role in circulation, by modulating the contractile responses of the arterial smooth muscle. This study was aimed at investigating the possible role of the endothelium in the contractile response to phorbol 12, 13-dibutyrate (PDB) in chronic two-kidney, one clip (2K1C) hypertensive rats. MATERIALS AND METHODS: 2K1C hypertension was induced by clipping the left renal artery of the study rats, with age-matched rats receiving a sham treatment, which served as controls. The thoracic aortae were mounted in tissue baths to measure the isometric tension. RESULTS: The PDB showed a dose-dependent contraction, with larger responses in the 2K1C hypertensive than the sham-clipped control rats. Nw-nitro-L-arginine (L-NNA) and methylene blue (MB) induced an increase in the tension in the presence of PDB, and the potentiating effects of L-NNA or MB were attenuated in the 2K1C rats as compared to the controls. Staurosporine, an inhibitor of protein kinase C, completely inhibited the contractile response to PDB, as well as enhancing the effects of L-NNA and MB. Removal of the endothelium abolished the contractile responses to L-NNA and MB in both the 2K1C and control rats. The relaxation responses to acetylcholine in the aortic rings precontracted with PDB were also attenuated in the 2K1C rats, and L-NNA prevented the effect of the acetylc-holine-induced relaxation. Indomethacin, glibenclamide and iberiotoxin did not affect the PDB responses in both the 2K1C and control rats. CONCLUSION: These results indicate the endothelium plays an inhibitory role against PDB-induced contraction in rat aortae, by releasing nitric oxide, and the inhibitory role of the endothelium is impaired in 2K1C renal hypertension.
Acetylcholine ; Animals ; Aorta ; Aorta, Thoracic ; Baths ; Endothelium ; Endothelium, Vascular* ; Glyburide ; Hypertension ; Hypertension, Renal ; Indomethacin ; Methylene Blue ; Muscle, Smooth ; Nitric Oxide ; Placebos ; Protein Kinase C ; Rats* ; Relaxation ; Renal Artery ; Staurosporine

OBJECTICE: To compare the use of conventional, real-time compound, and pulse-inversion harmonic imaging in the evaluation of breast nodules. MATERIALS AND METHODS: Fifty-two breast nodules were included in this study, conducted between May and December 2000, in which conventional, real-time compound, and pulse-inversion harmonic images were obtained in the same plane. Three radiologists, each blinded to the interpretations of the other two, evaluated the findings, characterizing the lesions and ranking the three techniques from grade 1, the worst, to grade 3, the best. Lesion conspicuity was assessed, and lesions were also characterized in terms of their margin, clarity of internal echotexture, and clarity of posterior echo pattern. The three techniques were compared using Friedman's test, and interobserver agreement in image interpretation was assessed by means of the intraclass correlation coefficient. RESULTS: With regard to lesion conspicuity, margin, and internal echotexture of the nodules, real-time compound imaging was the best technique (p < 0.05); in terms of posterior echo pattern, the best was pulse-inversion harmonic imaging (p < 0.05). Real-time compound and pulse inversion harmonic imaging were better than conventional sonography in all evaluative aspects. Interobserver agreement was greater than moderate. CONCLUSION: Real-time compound and pulse-inversion harmonic imaging procedures are superior to conventional sonography in terms of both lesion conspicuity and the further characterization of breast nodules. Real-time compound imaging is the best technique for evaluation of the margin and internal echotexture of nodules, while pulse-inversion harmonic imaging is very effective for the evaluation of the posterior echo patterns.
Breast Neoplasms/*ultrasonography ; Comparative Study ; Female ; Fibrocystic Disease of Breast/*ultrasonography ; Human ; Middle Age ; Ultrasonography, Mammary/*methods

BACKGROUND: The epidemiologic and clinical features of HIV infection/AIDS are different among various races, regions, and countries. To determine the epidemiologic and clinical characteristics of HIV infection in Korea, we analyzed and compared with that of other populations. METHODS: Medical records of 176 HIV-infected persons in Severance Hospital of Yonsei University College of Medicine and Hospital of Pusan University College of Medicine from year 1985 to 2000 were reviewed retrospectively. RESULTS: One hundred and seventy six patients were analyzed among which 156 (88.6%) were male and 20 (11.4%) were female with a male to female ratio of 7.8:1. At the time of diagnosis, the age distribution was 78 cases (44.3%) in the thirties, 44 cases (25.0%) in the twenties, and 35 cases (19.9%) in the fourties, and the mean age was 35.9+/-9.3. Heterosexual contact was the most frequent transmission route (92 cases, 52.3%), and 42 cases (23.9%) were transmitted by homosexual contact. At initial visit, asymptomatic HIV infection constituted 75 cases (42.6%), and AIDS 72 cases (40.9%). At initial visit, mean value of CD4+ lymphocyte counts was 252/mm3 and HIV RNA 226,035 copies/mm3. One hundred and twenty one of 176 patients developed 317 cases of opportunistic diseases. At the diagnosis of HIV-related opportunistic diseases, mean CD4+ lymphocyte count was 140/mm3 and mean HIV RNA 347,403 copies/mm3. Candidiasis (50 cases, 28.4%) was the most frequent opportunistic disease followed by pneumocystis carinii pneumonia (PCP) (37 cases, 21.0%), tuberculosis (29 cases, 16.5%), cytomegalovirus (CMV) infection (21 cases, 11.9%), HIV encephalopathy (9 cases, 5.1%), and herpes zoster (9 cases, 5.1%). There were 3 cases (1.7%) of malignant lymphoma and 2 cases (1.1%) of Kaposi's sarcoma. At the diagnosis of opportunistic diseases, mean CD4+ lymphocyte counts of patients with candidiasis was 71/mm3, PCP 63/mm3, and tuberculosis 142/mm3, and the mean HIV RNA level was 338,474 copies/mm3, 281,967 copies/mm3, and 817,012 copies/mm3 respectively. Among the 317 opportunistic diseases, AIDS-defining diseases were 150 cases (47.3%), of which PCP was 37 cases (24.7%), tuberculosis 29 cases (19.3%), CMV infection 21 cases (14.0%), HIV wasting syndrome 15 cases (10.0%), and esophageal candidiasis 14 cases (9.3%). The earliest AIDS-defining diseases to manifest in AIDS patients were tuberculosis (25 cases, 33.3%), followed by PCP (17 cases, 22.6%), esophageal candidiasis (14 cases, 18.7%), CMV infection (5 cases, 6.6%), and HIV wasting syndrome (4 cases, 5.3%). Thirty five (19.9%) of 176 patients were died. The common causes of death were tuberculosis (9 cases, 25.7%), PCP (9 cases, 25.7%), bacterial pneumonia (7 cases, 20.0%) and HIV encephalopathy (3 cases, 8.5%). CONCLUSION: The epidemiologic and clinical features of HIV infection/AIDS in Korea are different from that of developing countries such as Southeast Asia and Africa as well as from that of developed countries.
Africa ; Age Distribution ; AIDS Dementia Complex ; Asia, Southeastern ; Busan ; Candidiasis ; Cause of Death ; Continental Population Groups ; Cytomegalovirus ; Developed Countries ; Developing Countries ; Diagnosis ; Epidemiology ; Female ; Herpes Zoster ; Heterosexuality ; HIV Infections ; HIV Wasting Syndrome ; HIV* ; Homosexuality ; Humans ; Korea ; Lymphocyte Count ; Lymphoma ; Male ; Medical Records ; Pneumonia, Bacterial ; Pneumonia, Pneumocystis ; Retrospective Studies ; RNA ; Sarcoma, Kaposi ; Tuberculosis