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1.
Platelets ; 34(1): 2151996, 2022 12.
Article in English | MEDLINE | ID: mdl-36529914

ABSTRACT

Platelets are known to improve the wound-repair capacity of mesenchymal stem cells (MSCs) by transferring mitochondria intercellularly. This study aimed to investigate whether direct transfer of mitochondria (pl-MT) isolated from platelets could enhance wound healing in vitro using a cell-based model. Wound repairs were assessed by 2D gap closure experiment in wound scratch assay using human dermal fibroblasts (hDFs). Results demonstrated that pl-MT were successfully internalized into hDFs. It increased cell proliferation and promoted the closure of wound gap. Importantly, pl-MT suppressed both intracellular and mitochondrial ROS production induced by hydrogen peroxide, cisplatin, and TGF-ß in hDFs. Taken together, these results suggest that pl-MT transfer might be used as a potential therapeutic strategy for wound repair.


What is the context? During the wound healing process, abnormal regulation of ROS and inflammation delays the healing process, resulting in chronic non-healing wounds.Mitochondria are key organelles responsible for the ROS generation. Mitochondrial dysfunction has been implicated in delayed wound repair.Mitochondria transfer, which utilizes intact mitochondria isolated from healthy cells to recover from disease, has been applied in various clinical studies, but additional evidence is needed to apply it to wound healing.What is new? In this study, we chose platelets as a cell source for mitochondrial transfer. We isolated the functional mitochondria from platelets and applied them to wound healing.What is the impact? This study provides evidence that platelet-derived mitochondria (pl-MT) improve the wound healing progress by increasing the viability of dermal fibroblasts and suppressing intracellular and mitochondrial ROS production.Platelets have also been demonstrated to be a suitable cell source for mitochondrial transfer.


Subject(s)
Blood Platelets , Wound Healing , Humans , Blood Platelets/metabolism , Reactive Oxygen Species/metabolism , Fibroblasts , Mitochondria
2.
Int J Mol Sci ; 23(24)2022 Dec 12.
Article in English | MEDLINE | ID: mdl-36555376

ABSTRACT

Mitochondria are organelles that play a vital role in cellular survival by supplying ATP and metabolic substrates via oxidative phosphorylation and the Krebs cycle. Hence, mitochondrial dysfunction contributes to many human diseases, including metabolic syndromes, neurodegenerative diseases, cancer, and aging. Mitochondrial transfer between cells has been shown to occur naturally, and mitochondrial transplantation is beneficial for treating mitochondrial dysfunction. In this study, the migration of mitochondria was tracked in vitro and in vivo using mitochondria conjugated with green fluorescent protein (MTGFP). When MTGFP were used in a coculture model, they were selectively internalized into lung fibroblasts, and this selectivity depended on the mitochondrial functional states of the receiving fibroblasts. Compared with MTGFP injected intravenously into normal mice, MTGFP injected into bleomycin-induced idiopathic pulmonary fibrosis model mice localized more abundantly in the lung tissue, indicating that mitochondrial homing to injured tissue occurred. This study shows for the first time that exogenous mitochondria are preferentially trafficked to cells and tissues in which mitochondria are damaged, which has implications for the delivery of therapeutic agents to injured or diseased sites.


Subject(s)
Idiopathic Pulmonary Fibrosis , Mitochondria , Mice , Humans , Animals , Mitochondria/metabolism , Lung/metabolism , Idiopathic Pulmonary Fibrosis/metabolism , Fibroblasts/metabolism
3.
Materials (Basel) ; 15(19)2022 Oct 03.
Article in English | MEDLINE | ID: mdl-36234217

ABSTRACT

A simple and sensitive AuNP-coated magnetic beads (AMB)-based electrochemical biosensor platform was fabricated for bioassay. In this study, AuNP-conjugated magnetic particles were successfully prepared using biotin-streptavidin conjugation. The morphology and structure of the nanocomplex were characterized by scanning electron microscopy (SEM) with energy-dispersive X-ray analysis (EDX) and UV-visible spectroscopy. Moreover, cyclic voltammetry (CV) was used to investigate the effect of AuNP-MB on alkaline phosphatase (ALP) for electrochemical signal enhancement. An ALP-based electrochemical (EC) immunoassay was performed on the developed AuNP-MB complex with indium tin oxide (ITO) electrodes. Subsequently, the concentration of capture antibodies was well-optimized on the AMB complex via biotin-avidin conjugation. Lastly, the developed AuNP-MB immunoassay platform was verified with extracellular vesicle (EV) detection via immune response by showing the existence of EGFR proteins on glioblastoma multiforme (GBM)-derived EVs (108 particle/mL) spiked in human plasma. Therefore, the signal-enhanced ALP-based EC biosensor on AuNP-MB was favorably utilized as an immunoassay platform, revealing the potential application of biosensors in immunoassays in biological environments.

4.
BMB Rep ; 55(7): 361, 2022 Jul.
Article in English | MEDLINE | ID: mdl-35892133

ABSTRACT

[Erratum to: BMB Reports 2022; 55(3): 136-141, PMID: 34488927, PMCID: PMC8972135] The BMB Reports would like to correct in BMB Rep. 55(3):136-141, titled "Human umbilical cord mesenchymal stem cell-derived mitochondria (PN-101) attenuate LPS-induced inflammatory responses by inhibiting NFκB signaling pathway". This research was supported by NRF-2016R1A2B4007640 grant (to C-H Kim). Since grant number is incorrect, this information has now been corrected as follows: We would like to thank various Paean Biotechnology Inc. members who participated in the project. This work was supported by NRF-2018M3A9B5023055 grant (to C-H Kim). The authors apologize for any inconvenience or confusion that may be caused by this error. The ACKNOWLEDGEMENTS of Original PDF version have been corrected.

5.
BMB Rep ; 55(3): 136-141, 2022 Mar.
Article in English | MEDLINE | ID: mdl-34488927

ABSTRACT

Inflammation is one of the body's natural responses to injury and illness as part of the healing process. However, persistent inflammation can lead to chronic inflammatory diseases and multi-organ failure. Altered mitochondrial function has been implicated in several acute and chronic inflammatory diseases by inducing an abnormal inflammatory response. Therefore, treating inflammatory diseases by recovering mitochondrial function may be a potential therapeutic approach. Recently, mitochondrial transplantation has been proven to be beneficial in hyperinflammatory animal models. However, it is unclear how mitochondrial transplantation attenuates inflammatory responses induced by external stimuli. Here, we isolated mitochondria from umbilical cord-derived mesenchymal stem cells, referred as to PN-101. We found that PN-101 could significantly reduce LPS-induced mortality in mice. In addition, in phorbol 12-myristate 13-acetate (PMA)-treated THP-1 macrophages, PN-101 attenuated LPS-induced increase production of pro-inflammatory cytokines. Furthermore, the anti-inflammatory effect of PN-101 was mediated by blockade of phosphorylation, nuclear translocation, and trans-activity of NFκB. Taken together, our results demonstrate that PN-101 has therapeutic potential to attenuate pathological inflammatory responses. [BMB Reports 2022; 55(3): 136-141].


Subject(s)
Lipopolysaccharides , Mesenchymal Stem Cells , Animals , Cytokines/metabolism , Humans , Inflammation/chemically induced , Inflammation/metabolism , Lipopolysaccharides/pharmacology , Mesenchymal Stem Cells/metabolism , Mice , Mitochondria/metabolism , NF-kappa B/metabolism , Signal Transduction , Umbilical Cord/metabolism
6.
Int Immunopharmacol ; 64: 16-23, 2018 Nov.
Article in English | MEDLINE | ID: mdl-30144640

ABSTRACT

In our previous study, we demonstrated that sesamolin can increase the level of cancer cell susceptibility to natural killer (NK) cell mediated cytolysis when it treats cancer cells. The present study attempted to demonstrate the direct influence of sesamolin on NK cells. To achieve the study goal, an NK cell (NK-92MI) or Raji cell was treated with sesamolin for use in the analysis of the cytolytic activity of NK cells. When NK-92MI cells were treated with sesamolin, the cytolysis activities of NK cells increased depending on the concentration of sesamolin. However, the highest cytolytic activity of NK cells was observed when Raji and NK-92MI cells were treated with sesamolin at 20 µg/mL and 40 µg/mL, respectively. Sesamolin also increased the expression of the degranulation marker, CD107a, on the surface of NK cells and the production of immune-activation cytokine, IFN-γ, from NK cells. The effects of sesamolin on NK cells were reproduced in the naïve NK cells. We found that sesamolin effects are triggered by the result of phosphorylation of the p38, ERK1/2 and JNK pathways in NK cells. Taken together, this study proved that NK cell activity can be increased by the stimulation of sesamolin on NK cells as well as cancer cells.


Subject(s)
Dioxoles/pharmacology , Killer Cells, Natural/drug effects , Neoplasms/drug therapy , Cell Line, Tumor , Cell Survival/drug effects , Cytotoxicity, Immunologic/drug effects , Humans , Killer Cells, Natural/immunology , Lymphocyte Activation/drug effects , Lysosomal-Associated Membrane Protein 1/analysis , NK Cell Lectin-Like Receptor Subfamily K/analysis , Neoplasms/immunology
7.
Gut Liver ; 8(2): 140-7, 2014 Mar.
Article in English | MEDLINE | ID: mdl-24672654

ABSTRACT

BACKGROUND/AIMS: DA-9701, a standardized extract of Pharbitis Semen and Corydalis Tuber, is a new prokinetic agent that exhibits an analgesic effect on the abdomen. We investigated whether DA-9701 affects visceral pain induced by colorectal distension (CRD) in rats. METHODS: A total of 21 rats were divided into three groups: group A (no CRD+no drug), group B (CRD+no drug), and group C (CRD+DA-9701). Expression of pain-related factors, substance P (SP), c-fos, and phosphorylated extracellular signal-regulated kinase (p-ERK) in the dorsal root ganglion (DRG) and spinal cord was determined by immunohistochemical staining and Western blotting. RESULTS: The proportions of neurons in the DRG and spinal cord expressing SP, c-fos, and p-ERK were higher in group B than in group A. In the group C, the proportion of neurons in the DRG and spinal cord expressing p-ERK was lower than that in group B. Western blot results for p-ERK in the spinal cord indicated a higher level of expression in group B than in group A and a lower level of expression in group C than in group B. CONCLUSIONS: DA-9701 may decrease visceral pain via the downregulation of p-ERK in the DRG and spinal cord.


Subject(s)
Analgesics/pharmacology , Extracellular Signal-Regulated MAP Kinases/metabolism , Ganglia, Spinal/metabolism , Plant Preparations/pharmacology , Spinal Cord/metabolism , Animals , Colon , Dilatation, Pathologic/physiopathology , Down-Regulation , Extracellular Signal-Regulated MAP Kinases/drug effects , Ganglia, Spinal/drug effects , Male , Phytotherapy/methods , Proto-Oncogene Proteins c-fos/metabolism , Rats , Rats, Sprague-Dawley , Rectum , Spinal Cord/drug effects , Substance P/metabolism , Visceral Pain/prevention & control
8.
J Chromatogr Sci ; 52(8): 886-93, 2014 Sep.
Article in English | MEDLINE | ID: mdl-24023058

ABSTRACT

A sensitive and rapid method for quantitation of bepotastine in human plasma has been established using ultra performance liquid chromatography-electrospray ionization tandem mass spectrometry (UPLC-ESI-MS/MS). Valsartan was used as an internal standard. Bepotastine and internal standard in plasma sample were extracted using ethylacetate (liquid-liquid extraction). A centrifuged upper layer was then evaporated and reconstituted with the mobile phase of acetonitrile--5 mM ammonium formate (pH 3.5) (85:15, v/v). The reconstituted samples were injected into a phenyl column. Using MS/MS in the multiple reaction monitoring mode, bepotastine and valsartan were detected without severe interference from human plasma matrix. Bepotastine produced a protonated precursor ion ([M+H](+)) at m/z 389 and a corresponding product ion at m/z 167. And the internal standard produced a protonated precursor ion ([M+H](+)) at m/z 436 and a corresponding product ion at m/z 291. Detection of bepotastine in human plasma by the UPLC-ESI-MS/MS method was accurate and precise with a quantitation limit of 0.2 ng/mL. The validation, reproducibility, stability and recovery of the method were evaluated. The method has been successfully applied to pharmacokinetic studies of bepotastine in human plasma.


Subject(s)
Chromatography, Liquid/methods , Piperidines/blood , Pyridines/blood , Spectrometry, Mass, Electrospray Ionization/methods , Tandem Mass Spectrometry/methods , Administration, Oral , Drug Stability , Humans , Male , Piperidines/pharmacokinetics , Pyridines/pharmacokinetics , Reproducibility of Results
9.
Exp Mol Med ; 45: e53, 2013 Nov 15.
Article in English | MEDLINE | ID: mdl-24232252

ABSTRACT

The transplantation of neural precursor cells (NPCs) is known to be a promising approach to ameliorating behavioral deficits after stroke in a rodent model of middle cerebral artery occlusion (MCAo). Previous studies have shown that transplanted NPCs migrate toward the infarct region, survive and differentiate into mature neurons to some extent. However, the spatiotemporal dynamics of NPC migration following transplantation into stroke animals have yet to be elucidated. In this study, we investigated the fates of human embryonic stem cell (hESC)-derived NPCs (ENStem-A) for 8 weeks following transplantation into the side contralateral to the infarct region using 7.0T animal magnetic resonance imaging (MRI). T2- and T2*-weighted MRI analyses indicated that the migrating cells were clearly detectable at the infarct boundary zone by 1 week, and the intensity of the MRI signals robustly increased within 4 weeks after transplantation. Afterwards, the signals were slightly increased or unchanged. At 8 weeks, we performed Prussian blue staining and immunohistochemical staining using human-specific markers, and found that high percentages of transplanted cells migrated to the infarct boundary. Most of these cells were CXCR4-positive. We also observed that the migrating cells expressed markers for various stages of neural differentiation, including Nestin, Tuj1, NeuN, TH, DARPP-32 and SV38, indicating that the transplanted cells may partially contribute to the reconstruction of the damaged neural tissues after stroke. Interestingly, we found that the extent of gliosis (glial fibrillary acidic protein-positive cells) and apoptosis (TUNEL-positive cells) were significantly decreased in the cell-transplanted group, suggesting that hESC-NPCs have a positive role in reducing glia scar formation and cell death after stroke. No tumors formed in our study. We also performed various behavioral tests, including rotarod, stepping and modified neurological severity score tests, and found that the transplanted animals exhibited significant improvements in sensorimotor functions during the 8 weeks after transplantation. Taken together, these results strongly suggest that hESC-NPCs have the capacity to migrate to the infarct region, form neural tissues efficiently and contribute to behavioral recovery in a rodent model of ischemic stroke.


Subject(s)
Cell Movement , Embryonic Stem Cells/transplantation , Infarction, Middle Cerebral Artery/surgery , Neural Stem Cells/transplantation , Psychomotor Performance , Animals , Apoptosis , Cell Differentiation , Embryonic Stem Cells/cytology , Embryonic Stem Cells/metabolism , Glial Fibrillary Acidic Protein/genetics , Glial Fibrillary Acidic Protein/metabolism , Humans , Infarction, Middle Cerebral Artery/metabolism , Infarction, Middle Cerebral Artery/pathology , Infarction, Middle Cerebral Artery/physiopathology , Male , Neural Stem Cells/cytology , Neural Stem Cells/metabolism , Rats , Rats, Sprague-Dawley , Receptors, CXCR4/genetics , Receptors, CXCR4/metabolism
10.
Neurosci Lett ; 548: 291-5, 2013 Aug 26.
Article in English | MEDLINE | ID: mdl-23769729

ABSTRACT

It has been suggested that activation of alpha7 nicotinic acetylcholine receptors (α7nAChR) could alleviate acute and chronic pain in various abnormal pain models. However, it is unclear whether the stimulation of α7nAChRs has anti-hyperalgesic effects on osteoarthritis. Therefore, we tested whether choline, an α7nAChR agonist, could alleviate chronic inflammatory pain in an osteoarthritis model. Osteoarthritis was induced by injection of monoiodoacetic acid (MIA) into the synovial cavity of the knee joints in rats. Pain was assessed by responses to stimuli on the plantar surface: paw withdrawal threshold (PWT) by up-down methods using a series of von Frey filaments, and paw withdrawal latency (PWL) using radiation heat. Both PWT and PWL decreased after MIA injection, indicating development of mechanical and thermal hyperalgesia. Subsequent intraperitoneal choline injection increased both PWT and PWL. PWT increased in response to choline injections (5-50 mg/Kg) in a dose dependent manner. PWL increased significantly in a similar fashion in response to choline (20 and 50 mg/Kg). However, intraarticular injection of choline did not result in any change in PWT or PWL. Intrathecal choline increased PWT and PWL. The anti-hyperalgesic effect of intraperitoneal choline was completely blocked by methyllycaconitine when it was injected intrathecally 10 min before the choline treatment. These results show that choline could alleviate mechanical and heat hyperalgesia via spinal α7nAChR in the MIA-induced inflammation pain model.


Subject(s)
Arthralgia/drug therapy , Choline/administration & dosage , Hyperalgesia/drug therapy , Osteoarthritis/drug therapy , Spinal Cord/metabolism , alpha7 Nicotinic Acetylcholine Receptor/agonists , alpha7 Nicotinic Acetylcholine Receptor/metabolism , Animals , Arthralgia/etiology , Arthralgia/metabolism , Hyperalgesia/etiology , Hyperalgesia/metabolism , Male , Osteoarthritis/complications , Osteoarthritis/metabolism , Rats , Rats, Sprague-Dawley , Spinal Cord/drug effects , Treatment Outcome
11.
Dig Dis Sci ; 57(6): 1517-24, 2012 Jun.
Article in English | MEDLINE | ID: mdl-22427128

ABSTRACT

BACKGROUND AND AIMS: Mosapride citrate is known to affect gastric motility. However, whether mosapride citrate has any effect on visceral pain in the colon or rectum is not certain. The aim of this study was to assess the effects of mosapride citrate on visceral pain in a rat visceral hypersensitivity model. METHODS: The perception of visceral pain was evaluated by the visceromotor response to colorectal distension observed on electromyographs of the abdominal musculature in urethane-anesthetized rats. Visceral hypersensitivity was induced by the intrarectal instillation of 4% acetic acid or 1.5% zymosan. Mosapride citrate was administered intraperitoneally 3 h later. VMRs to CRD were recorded prior to the instillation of acetic acid or zymosan and before and after mosapride citrate treatment. RESULTS: The intracolonic instillation of acetic acid resulted in a significant increase in VMRs of the abdominal muscles to CRD, compared with the pretreatment state (174 ± 24%, P < 0.05). The intracolonic instillation of zymosan resulted in a significant increase in VMRs of the abdominal muscles to CRD, compared with the pretreatment state (144 ± 9%, P < 0.05). Intraperitoneal injection of mosapride citrate resulted in a significant reduction in the VMRs to CRD in an acetic acid-induced visceral hypersensitivity rat model (61 ± 9%, P < 0.05). The intraperitoneal injection of mosapride citrate also resulted in a significant reduction in the VMRs to CRD in a zymosan-induced visceral hypersensitivity rat model (67 ± 9%, P < 0.05). CONCLUSIONS: Mosapride citrate diminished visceral pain in rats.


Subject(s)
Benzamides/pharmacology , Gastrointestinal Motility/drug effects , Morpholines/pharmacology , Serotonin 5-HT4 Receptor Agonists/pharmacology , Visceral Pain/drug therapy , Animals , Disease Models, Animal , Electromyography , Gastrointestinal Motility/physiology , Hypersensitivity/drug therapy , Hypersensitivity/physiopathology , Injections, Intraperitoneal , Male , Pain Measurement , Random Allocation , Rats , Rats, Sprague-Dawley , Reference Values , Treatment Outcome , Visceral Pain/physiopathology
12.
Neurosci Lett ; 500(3): 197-201, 2011 Aug 18.
Article in English | MEDLINE | ID: mdl-21742015

ABSTRACT

Group 1 metabotropic glutamate receptors (mGluRs) are expressed in peripheral and central neural tissues and involved in peripheral and central sensitization in various pain models. However, there are limited reports that activation of peripheral group I mGluRs could evoke pain. Furthermore, any behavioral evidences could not be found out, showing what kind of afferent fibers are involved in peripheral mGluRs-mediated hyperalgesia. This study was undertaken to clarify whether peripherally injected group I mGluRs agonists could induce pain-related behaviors and capsaicin-sensitive afferent fibers might be involved in the hyperalgesia. To assess pain sensitivity, mechanical threshold for paw withdrawal response (PWT) was measured and number of spontaneous flinching behavior was counted. Intraplantar injection of group I mGluR agonist, (RS)-3,5-dihydroxyphenylglycine (DHPG) and mGluR5 agonist, (RS)-2-chloro-5-hydroxyphenyglycine (CHPG) immediately induced pain-like behaviors, such as decrease of PWT and increased number of flinchings. These agonists-induced pain-like behaviors were blocked by group I mGluRs antagonist, (RS)-1-aminoindan-1,5-dicarboxylic acid (AIDA) and mGluR5 antagonist, 2-methyl-6-(phenylethynyl) pyridine hydrochloride (MPEP). Perineural pretreatment of 1% capsaicin solution significantly reduced pain-related behaviors induced by DHPG and CHPG, proposing that capsaicin-sensitive primary afferent fibers could be responsible for the hyperalgesia induced by activation of peripheral group I mGluRs. This study presents the first behavioral evidence that peripheral group I mGluRs activation could induce spontaneous as well as mechanical hyperalgesia and capsaicin-sensitive afferent fiber could be implicated the group I mGluR mediated hyperalgesia.


Subject(s)
Capsaicin/pharmacology , Hyperalgesia/physiopathology , Receptors, Metabotropic Glutamate/agonists , Afferent Pathways , Animals , Hyperalgesia/metabolism , Male , Nerve Fibers/drug effects , Nerve Fibers/physiology , Pain Threshold/drug effects , Rats , Rats, Sprague-Dawley , Receptors, Metabotropic Glutamate/antagonists & inhibitors , Sciatic Nerve/drug effects , Sciatic Nerve/physiopathology , Touch
13.
Surg Radiol Anat ; 32(3): 235-8, 2010 Mar.
Article in English | MEDLINE | ID: mdl-19937327

ABSTRACT

The suprascapular nerve branches provide efferent innervation to the supraspinatus and infraspinatus muscles as well as sensory innervation to the shoulder joint. This study was carried out to verify the spinal root origins and innervations of the suprascapular nerve. Fifty samples of the suprascapular nerve taken from 37 adult Korean cadavers were used in this study. The suprascapular nerve was found to comprise the ventral rami of the C5 and C6 in 76.0% of the fifty samples; C4, C5, and C6 nerves in 18.0%; and C5 nerve in only 6.0%. The C5 nerve was consistently shown to be the largest in mean diameter and was found to be a major contributor of nerve fibers leading to the suprascapular nerve. This study shows that the main spinal component of the suprascapular nerve is C5 nerve. In most cases, the rate of the involvement of the C4 and C6 nerves (18.0 and 94.0%, respectively) with the suprascapular nerve was less than that of C5 nerve. C4 and C5 nerves were shown to contribute nerve fibers to the supraspinatus and infraspinatus muscles and to both shoulder joints, whereas C6 nerve displayed variable patterns of innervation.


Subject(s)
Brachial Plexus/anatomy & histology , Muscle, Skeletal/innervation , Shoulder Joint/innervation , Spinal Nerve Roots/anatomy & histology , Aged , Cadaver , Female , Humans , Male , Shoulder Joint/anatomy & histology
14.
Neurosci Lett ; 433(3): 199-204, 2008 Mar 15.
Article in English | MEDLINE | ID: mdl-18242851

ABSTRACT

Whether modulation of C afferent fiber activities could relieve peripheral neuropathic pain was tested. After establishment of neuropathic pain induced by L5 and 6 spinal nerve transection (SNT), the sciatic nerve was treated with 2% capsaicin at the level of the midthigh. Mechanical hyperalgesia (von Frey filaments) was significantly alleviated from 7 days to 4 weeks after capsaicin treatment, but cold allodynia (acetone) was unchanged. Immunohistochemical studies showed a significant increase in the number of calcitonin gene-related peptide (CGRP)-positive neurons, but not TRPV1-positive neurons in intact L4 dorsal root ganglia after SNT. Capsaicin treatment decreased TRPV1- and CGRP-positive neurons in L4 DRG of the treated side, but not the opposite side. These results suggest that local application of capsaicin onto the sciatic nerve can alleviate mechanical hyperalgesia, but not cold allodynia, in a peripheral neuropathic pain model and the pain alleviation may result from a decrease of TRPV1- and CGRP-positive sensory neurons of which fibers pass through the sciatic nerve.


Subject(s)
Capsaicin/pharmacology , Hyperalgesia/drug therapy , Neuralgia/drug therapy , Peripheral Nerve Injuries , Peripheral Nerves/drug effects , Peripheral Nervous System Diseases/drug therapy , Analgesics/pharmacology , Animals , Biomarkers/analysis , Biomarkers/metabolism , Calcitonin Gene-Related Peptide/metabolism , Cell Count , Denervation , Ganglia, Spinal/drug effects , Ganglia, Spinal/metabolism , Ganglia, Spinal/physiopathology , Hyperalgesia/metabolism , Hyperalgesia/physiopathology , Male , Nerve Degeneration/metabolism , Nerve Degeneration/physiopathology , Neuralgia/metabolism , Neuralgia/physiopathology , Neurons, Afferent/drug effects , Neurons, Afferent/metabolism , Pain Measurement , Peripheral Nerves/physiopathology , Peripheral Nervous System Diseases/metabolism , Peripheral Nervous System Diseases/physiopathology , Physical Stimulation , Rats , Rats, Sprague-Dawley , Sensory System Agents/pharmacology , TRPV Cation Channels/metabolism
15.
BMC Bioinformatics ; 8: 485, 2007 Dec 20.
Article in English | MEDLINE | ID: mdl-18096047

ABSTRACT

BACKGROUND: The quality of cDNA microarray data is crucial for expanding its application to other research areas, such as the study of gene regulatory networks. Despite the fact that a number of algorithms have been suggested to increase the accuracy of microarray gene expression data, it is necessary to obtain reliable microarray images by improving wet-lab experiments. As the first step of a cDNA microarray experiment, spotting cDNA probes is critical to determining the quality of spot images. RESULTS: We developed a governing equation of cDNA deposition during evaporation of a drop in the microarray spotting process. The governing equation included four parameters: the surface site density on the support, the extrapolated equilibrium constant for the binding of cDNA molecules with surface sites on glass slides, the macromolecular interaction factor, and the volume constant of a drop of cDNA solution. We simulated cDNA deposition from the single model equation by varying the value of the parameters. The morphology of the resulting cDNA deposit can be classified into three types: a doughnut shape, a peak shape, and a volcano shape. The spot morphology can be changed into a flat shape by varying the experimental conditions while considering the parameters of the governing equation of cDNA deposition. The four parameters were estimated by fitting the governing equation to the real microarray images. With the results of the simulation and the parameter estimation, the phenomenon of the formation of cDNA deposits in each type was investigated. CONCLUSION: This study explains how various spot shapes can exist and suggests which parameters are to be adjusted for obtaining a good spot. This system is able to explore the cDNA microarray spotting process in a predictable, manageable and descriptive manner. We hope it can provide a way to predict the incidents that can occur during a real cDNA microarray experiment, and produce useful data for several research applications involving cDNA microarrays.


Subject(s)
Image Interpretation, Computer-Assisted/methods , In Situ Hybridization, Fluorescence/methods , Microfluidics/methods , Microscopy, Fluorescence/methods , Models, Chemical , Models, Genetic , Oligonucleotide Array Sequence Analysis/methods , Computer Simulation , Reproducibility of Results , Sensitivity and Specificity
16.
Arzneimittelforschung ; 57(8): 542-6, 2007.
Article in English | MEDLINE | ID: mdl-17915642

ABSTRACT

The aims of this study were to assess the pharmacokinetics and bioequivalence of two brands of loxoprofen (CAS 80832-23-6) 60 mg tablets in healthy male volunteers. The several pharmacokinetic parameters were evaluated after an oral administration after an overnight fast according to a single dose, two-sequence, and cross-over randomized design with a 1-week washout interval. Serial blood samples were collected throughout 10 h after administration of the reference and test drug. Plasma was analyzed by validated HPLC with UV detection. Several pharmacokinetic parameters, including AUC(infnity), AUC(t), C(max), T(max), T1/2, and Ke were determined from blood concentrations of both formulations. AUC(t), AUC(infinity) and C(max) were evaluated for bioequivalence after log-transformation of data using ANOVA with 90% confidence interval level. The parametric 90% confidence intervals of AUC(t), AUC(infinity), and C(max) were 90.13-106.34%, 91.43-106.94%, and 91.17-108.53%, respectively. All of the tested parameters were within the acceptable range of 80-125%. Based on these statistical considerations, it was concluded that the test drug was bioequivalent to the reference drug.


Subject(s)
Anti-Inflammatory Agents, Non-Steroidal/pharmacokinetics , Phenylpropionates/pharmacokinetics , Adult , Anti-Inflammatory Agents, Non-Steroidal/administration & dosage , Area Under Curve , Calibration , Chromatography, High Pressure Liquid , Cross-Over Studies , Half-Life , Humans , Indicators and Reagents , Male , Phenylpropionates/administration & dosage , Reproducibility of Results , Spectrophotometry, Ultraviolet , Tablets
17.
Arzneimittelforschung ; 57(7): 455-61, 2007.
Article in English | MEDLINE | ID: mdl-17803058

ABSTRACT

A rapid and validated liquid chromatography coupled to tandem mass spectrometric method (LC-MS-MS) has been developed and applied to pharmacokinetic and bioequivalence studies in 24 healthy male Korean volunteers. The procedure involves a liquid-liquid extraction of paroxetine (CAS 61869-08-7) and fluoxetine (internal standard, CAS 54910-89-3) with ether/methyl chloride (7:3, v/v) and separated by LC equipped with C18 column using acetonitrile: 5 mmol/L ammonium formate (4:3, v/v) as mobile phase. Detection is carried out on an API 2000 MS system by multiple reactions monitoring mode. The ionization was optimized using ESI(+) and selectivity was achieved by MS-MS analysis, mlz 330.0-->192.0 and m/ z 310-->148 for paroxetine and fluoxetine, respectively. The method has a total run time of 1.5 min and was linear over a working range of 0.05-20 ng/mL and the lower limit of quantification was 0.05 ng/ mL. No endogenous compounds were found to interfere with the analysis. The inter-day and intra-day accuracy was in the ranges of 102.69-107.79% and 102.07-109.57%, respectively and precision of inter-day and intra-day expressed as relative standard deviation were 1.86-9.99% and 1.52-6.28%, respectively. The validation of this method on linearity, specificity, accuracy, precision as well as applicability to pharmacokinetic and bioequivalence studies by analysis of blood samples taken up to 72 h after oral administration of 20 mg of paroxetine in 24 healthy volunteers were found to be good performance.


Subject(s)
Antidepressive Agents, Second-Generation/blood , Antidepressive Agents, Second-Generation/pharmacokinetics , Paroxetine/blood , Paroxetine/pharmacokinetics , Area Under Curve , Calibration , Chromatography, High Pressure Liquid , Chromatography, Liquid , Cross-Over Studies , Humans , Reference Standards , Reproducibility of Results , Solutions , Tablets , Tandem Mass Spectrometry , Therapeutic Equivalency
18.
Neurosci Res ; 58(3): 245-9, 2007 Jul.
Article in English | MEDLINE | ID: mdl-17428562

ABSTRACT

Recently, it has been suggested that uninjured primary sensory neurons contribute to neuropathic pain induced by peripheral nerve injury. However, there is lack of evidences of roles of normal pain transmitting substances such as substance P and calcitonin gene-related peptide (CGRP) in neuropathic pain. Whether substance P and CGRP have a role in spinal nerve-injured neuropathic pain model was tested. Male rats were subjected to L5 and L6 spinal nerve transection (SNT), and mechanical hyperalgesia was evaluated by measuring paw withdrawal threshold (PWT). SNT induced a persistent PWT decrease, a sign of neuropathic pain. Lidocaine was soaked on spinal nerves or intrathecally injected 10 min before SNT to block neuronal discharges caused by the injury, and L703,606 (NK1 receptor antagonist) and CGRP8-37 (CGRP receptor antagonist) were intrathecally injected into the rats to block actions of substance P and CGRP released from central nerve terminals in the spinal cord by injury discharges. The treatments with lidocaine, L703,606 and CGRP8-37 delayed the onset of neuropathic pain by 1-4 days, compared with the saline-treated rats. After neuropathic pain was established, intrathecal injections of L703,606 and CGRP8-37 significantly mitigated mechanical hyperalgesia for 20 min. These results suggest that substance P and CGRP are involved in the development and maintenance of neuropathic pain and that these peptides from the central terminals of intact sensory neurons contribute to the maintenance of peripheral nerve injury-induced neuropathic pain.


Subject(s)
Calcitonin Gene-Related Peptide/physiology , Neuralgia/metabolism , Neuralgia/pathology , Spinal Nerves , Substance P/physiology , Analysis of Variance , Anesthetics, Local/therapeutic use , Animals , Behavior, Animal , Calcitonin Gene-Related Peptide/therapeutic use , Disease Models, Animal , Dose-Response Relationship, Drug , Lidocaine/therapeutic use , Male , Neuralgia/drug therapy , Pain Measurement/methods , Peptide Fragments/therapeutic use , Quinuclidines/pharmacology , Rats , Rats, Sprague-Dawley , Time Factors
19.
Pain ; 94(2): 169-175, 2001 Nov.
Article in English | MEDLINE | ID: mdl-11690730

ABSTRACT

Intradermal injection of capsaicin induces primary hyperalgesia at the injection site and secondary hyperalgesia in the surrounding undamaged skin. The secondary hyperalgesia is thought to be due to central sensitization of the dorsal horn neurons while primary hyperalgesia is caused by sensitization of nociceptors in the damaged skin. In this study, we asked if additional non-noxious afferent input from the undamaged skin influences the already developed secondary hyperalgesia, which follows an intradermal injection of capsaicin. Capsaicin dissolved in olive oil was injected into the middle of the hind paw of male Sprague-Dawley rats (250-300 g) under gaseous anesthesia. This produced a decrease in the mechanical threshold at the base of the toes for hind limb withdrawals lasting for 1-2h, thus showing a short-lasting (hours) secondary hyperalgesia. When the capsaicin injection was immediately followed by repeated non-noxious mechanical stimuli or weak electrical stimuli (A fiber strength) applied to the area of secondary hyperalgesia (toes) for 30 min, the reduction of the mechanical threshold lasted longer than 24h. These results suggest that non-noxious A fiber afferent input can powerfully modulate central sensitization in the spinal dorsal horn, causing the duration of the secondary hyperalgesia to be greatly extended.


Subject(s)
Capsaicin/pharmacology , Hyperalgesia/physiopathology , Nerve Fibers, Myelinated/physiology , Animals , Behavior, Animal/drug effects , Electric Stimulation , Hyperalgesia/chemically induced , Male , Nerve Fibers, Myelinated/drug effects , Neuralgia/chemically induced , Neuralgia/physiopathology , Physical Stimulation , Rats , Rats, Sprague-Dawley
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