• Title/Summary/Keyword: 자극-반응법

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Extraction of Nature Pigment with Antioxidant Properties from Sprout Barley - Optimization Using CCD-RSM (새싹보리로부터 항산화기능성을 갖는 천연색소의 추출 - CCD-RSM을 이용한 최적화)

  • Dong Hwan Kim;Seung Bum Lee
    • Applied Chemistry for Engineering
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    • v.35 no.3
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    • pp.222-229
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    • 2024
  • The use of low-toxic, hypoallergenic, and environmentally friendly natural pigments has increased. With growing interest in health, research on natural extracts containing beneficial substances for the human body is actively underway. In this study, natural pigments were extracted from sprout barley using a solvent extraction method and CCD-RSM was used to optimize the extraction process. The experiment's independent variables included extraction temperature, alcohol/ultra-pure volume ratio, and extraction time. The response variables were set to achieve a target chromaticity (L = 45, a = -35, b = 45), and to maximize DPPH radical scavenging activity evaluating the antioxidant capacity. The statistical significance of the main effect, interaction effect, and effect on the response value was evaluated and analyzed through the F and P values for the regression equation variables calculated using RSM optimization. Additionally, the reliability of the experiment was also confirmed through the P values of the probability plot graph. The extraction conditions for optimizing the four reaction values are 76.1 vol.% alcohol/ultra pure water volume ratio, an extraction temperature of 52.9 ℃ , and an extraction time of 49.6 min. Under these conditions, the theoretical values of the reaction values are L = 45.4, a = -36.8, and b = 45.0 DPPH radical scavenging activity = 30.9%. When the actual experiment was conducted under these optimal extraction conditions and analyzed, the measured values were L = 46.2, a = -36.1, and b = 48.2, and antioxidant capacity = 31.1% with an average error rate of 2.9%.

Lipopolysaccharide-induced Synthesis of IL-1beta, IL-6, TNF-alpha and TGF-beta by Peripheral Blood Mononuclear Cells (내독소에 의한 말초혈액 단핵구의 IL-1beta, IL-6, TNF-alpha와 TGF-beta 생성에 관한 연구)

  • Jung, Sung-Hwan;Park, Choon-Sik;Kim, Mi-Ho;Kim, Eun-Young;Chang, Hun-Soo;Ki, Shin-Young;Uh, Soo-Taek;Moon, Seung-Hyuk;Kim, Yang-Hoon;Lee, Hi-Bal
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.4
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    • pp.846-860
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    • 1998
  • Background: Endotoxin (LPS : lipopolysaccharide), a potent activator of immune system, can induce acute and chronic inflammation through the production of cytokines by a variety of cells, such as monocytes, endothelial cells, lymphocytes, eosinophils, neutrophils and fibroblasts. LPS stimulate the mononucelar cells by two different pathway, the CD14 dependent and independent way, of which the former has been well documented, but not the latter. LPS binds to the LPS-binding protein (LBP), in serum, to make the LPS-LBP complex which interacts with CD14 molecules on the mononuclear cell surface in peripheral blood or is transported to the tissues. In case of high concentration of LPS, LPS can stimulate directly the macrophages without LBP. We investigated to detect the generation of proinflammatory cytokines such as interleukin 1 (IL-1), IL-6 and TNF-$\alpha$ and fibrogenic cytokine, TGF-$\beta$, by peripheral blood mononuclear cells (PBMC) after LPS stimulation under serum-free conditions, which lacks LBPs. Methods : PBMC were obtained by centrifugation on Ficoll Hypaque solution of peripheral venous bloods from healthy normal subjects, then stimulated in the presence of LPS (0.1 ${\mu}g/mL$ to 100 ${\mu}g/mL$ ). The activities of IL-1, IL-6, TNF, and TGF-$\beta$ were measured by bioassaies using cytokines - dependent proliferating or inhibiting cell lines. The cellular sources producing the cytokines was investigated by immunohistochemical stains and in situ hybridization. Results : PBMC started to produce IL-6, TNF-$\alpha$ and TGF-$\beta$ in 1 hr, 4 hrs and 8hrs, respectively, after LPS stimulation. The production of IL-6, TNF-$\alpha$ and TGF-$\beta$ continuously increased 96 hrs after stimulation of LPS. The amount of production was 19.8 ng/ml of IL-6 by $10^5$ PBMC, 4.1 ng/mL of TNF by $10^6$ PBMC and 34.4 pg/mL of TGF-$\beta$ by $2{\times}10^6$ PBMC. The immunoreactivity to IL-6, TNF-$\alpha$ and TGF-$\beta$ were detected on monocytes in LPS-stimulated PBMC. Some of lymphocytes showed positive immunoreactivity to TGF-$\beta$. Double immunohistochemical stain showed that IL-1$\beta$, IL-6, TNF-$\alpha$ expression was not associated with CD14 postivity on monocytes. IL-1$\beta$, IL-6, TNF-$\alpha$ and TGF-$\beta$mRNA expression were same as observed in immunoreactivity for each cytokines. Conclusion: When monocytes are stimulated with LPS under serum-free conditions, IL-6 and TNF-$\alpha$ are secreted in early stage of inflammation. In contrast, the secretion of TGF-$\beta$ arise in the late stages and that is maintained after 96 hrs. The main cells releasing IL-1$\beta$, IL-6, TNF-$\alpha$ and TGF-$\beta$ are monocytes, but also lymphocytes can secret TGF-$\beta$.

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A Comparative Analysis of GBEF According to Image Aquisition Method in Hepatobiliary Scan (간담도스캔의 영상수집방법에 따른 담즙배출율의 비교분석)

  • Kim, Yeong-Seon;Seo, Myeong-Deok;Lee, Wan-Kyu;Song, Jae-Beom
    • The Korean Journal of Nuclear Medicine Technology
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    • v.18 no.2
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    • pp.8-16
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    • 2014
  • Purpose The quantitative analysis of gallbladder emptying is very important in diagnosis of motility disorder of gallbladder and in biliary physiology. The GBEF obtain the statics aquisition method or the dynamic acquisition method in two ways. The purpose of this study is to compare the GBEF value of statics acquisition method and the dynamic acquisition method. And we find the best way for calculate GBEF. Materials and Methods The quantitative hepatobiliary scan with $^{99m}Tc$-mebrofenin was performed of 27 patients. Initial images were acquired statically, for 60 min after injection of the radioactive tracer. And if the gallbladder is visualized to 60 min, performed stimulation of gallbladder (1egg, 200 mL milk). After that, started acquisition of dynamic image for 30 min. After that, image of after fatty meal of the statics method were acquired on equal terms with 60 min image. The statics GBEF was calculated using the images of before fatty meal and post fatty meal by the statics method. The dynamic GBEF was calculated using the images of time of maximum bile juice uptake ($T_{max}$) and time of minimum bile juice uptake ($T_{min}$) images from the gallbladder time-activity curve. A bile juice is secreted from gallbladder while eating a fatty meal. that is named early GBEF and that was calculated using before fatty meal image of the statics method and 1 min image of the dynamic method. Results The result saw very big difference between two according to $T_{max}$. The result, were as follows. 1) In case of less than 1 min, the dynamic mean GBEF was $40.1{\pm}21.7%$, the statics mean GBEF was $51.5{\pm}23.6%$ in 16 cases. The early mean GBEF was $14.0{\pm}29.1%$. The GBEF of statics method was higher because that include secreted bile juice while performed stimulation of gallbladder. A difference of GB counts according to acquisition method and the early bile juice counts was $17.6{\pm}14.8%$ and $13.5{\pm}15.3%$. 2) In case of exceed than 1 min, the dynamic mean GBEF was $31.0{\pm}19.7%$, the statics mean GBEF was $21.3{\pm}19.4%$ in 7 cases. The early GBEF was $-6.9{\pm}4.9%$. The GBEF of dynamic method was higher because that include concentrated bile juice to $T_{max}$. A difference of GB counts according to acquisition method and the early bile juice counts was $14.3{\pm}7.3%$ and $5.9{\pm}3.9%$. Conclusion The statics method is very easy and simple, but in case of $T_{max}$ delay, the GBEF can be lower. The dynamic method is able to calculate accurately in case of $T_{max}$ delay, but in case of $T_{max}$ is less than 1 min, the GBEF can be lower because dynamic GBEF exclude secreted bile juice while performed stimulation of gallbladder. The best way to calculate GBEF is to scan with dynamic method preferentially and to choose suitable method between the two way after conform $T_{max}$ on the T-A curve of the dynamic method.

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Toll-like receptor 2 expression on monocytes from patients with BCG vaccine-associated suppurative lymphadenitis (BCG 접종 후 발생한 화농성 림프절염 환자의 단핵구에서 Toll-like receptor 2의 발현)

  • Oh, Hyun Joo;Shin, Kyung-Sue
    • Clinical and Experimental Pediatrics
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    • v.52 no.6
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    • pp.667-673
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    • 2009
  • Purpose : Toll-like receptor 2 (TLR2) is critical in the immune response to mycobacterial infections. The purpose of this study was to analyze TLR2 surface expressions and TLR2-mediated tumor necrosis factor-alpha ($TNF-{\alpha}$) and interleukin-6 (IL-6) production in patients with BCG vaccine-associated suppurative lymphadenitis. Methods : Peripheral monocytes were separated from 16 patients with BCG vaccine-associated suppurative lymphadenitis and 10 healthy controls using a magnet cell isolation kit. Monocytes ($1{\times}10^6$ cells/well) were incubated with a constant amount of $Pam_3CSK_4$ ($100{\mu}g/mL$) for 24 hours. TLR2 surface expression on monocytes was analyzed by FACS analysis and TLR-2 mRNA expression was determined by RT-PCR. TLR2-mediated $TNF-{\alpha}$ and IL-6 production were measured by ELISA. Results : In patients with BCG vaccine-associated suppurative lymphadenitis, low TLR2 expression on monocytes ($3.39{\pm}$1.2% versus $4.64{\pm}2.6%$) together with significantly lower TLR2 mRNA expression than in the healthy controls was seen after $Pam_3CSK_4$ stimulation. TLR2-mediated $TNF-{\alpha}$ and IL-6 production in patients with BCG vaccine-associated suppurative lymphadenitis ($TNF-{\alpha}$, $775.5{\pm}60.8pg/mL$; IL-6, $4,645.8{\pm}583.9pg/mL$) were also lesser than that in healthy controls ($TNF-{\alpha}$, $1,098.5{\pm}94.3pg/mL$; IL-6, $6,696.3{\pm}544.3pg/mL$). Conclusion : These findings suggest that low TLR2 expression on monocytes might be associated with increased susceptibility to BCG vaccine-associated suppurative lymphadenitis.

Difference in Isoproterenol-Stimulated Cyclic AMP Levels of Lymphocytes between the Subgroups of Depressive Disorders (우울장애 아형간에 Isoproterenol 자극후 임파구 Cyclic AMP Levels의 차이)

  • Bahk, Won-Myong;Jeon, Yang-Whan;Kim, Kwang-Soo;Sung, Yang-Sook
    • Korean Journal of Biological Psychiatry
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    • v.3 no.1
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    • pp.46-50
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    • 1996
  • This study was designed to examine the basal cyclic AMP levels and the $10^{-5}mol/L$ isoproterenol-stimulated cyclic AMP levels of lymphocytes, by which ${\beta}$-adrenoceptor function was shown, between to normal controls and 17 drug free patients(8 major depresive patients and 9 dysthymic patients), who were diagnosed by DSM-III-R. The severity of depression was assessed by Hamilton Rating Scale for Depression (HDRS). Cyclic AMP levels were measured by radioimmunoassay(double antibody). The results were as follows ; 1) HDRS score was significantly higher in major depressive patients($41.8{\pm}4.6$) than in dysthymic patients($24.0{\pm}4.2$)(p<005). 2) There was no Significant difference in basal cyclic AMP levels among normal controls($3.9{\pm}1.7pmol/10^6cells/10min$), major depressive patients($2.1{\pm}0.5pmol/10^6cells/10min$), and dysthymic patients($3.9{\pm}1.8pmol/10^6cells/10min$). 3) There was significant difference in net cyclic AMP levels($10^{-5}mol/L$ isoproterenol-stimulated cyclic AMP levels minus basal cyclic AMP levels) among normal controls($16.5{\pm}6.0pmol/10^6cells/10min$), major depressive patients($3.0{\pm}1.4pmol/10^6cells/10min$), dysthymic patients($10.9{\pm}4.4pmol/10^6cells/10min$)(p <005). 4) The net cyclic AMP levels were significantly correlated with HDRS scores in major depressive patients(${\gamma}=-0.8^6$, p<0.05), but not in dysthymic patients(${\gamma}=0.43$, p=0.25). In conclusion, we suggested that the dysthymic disorder might differ from the molar depressive disorder not only in the severity of depressive symptoms but also in ${\beta}$-adrenergic responsiveness of lymphocytes, which was regarded as a biological marker of depressive disorder.

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Optimization of a Crystallization Process by Response Surface Methodology (반응표면분석법을 이용한 결정화 공정의 최적화)

  • Lee, Se-Eun;Kim, Jae-Kyeong;Han, Sang-Keun;Chae, Joo-Seung;Lee, Keun-Duk;Koo, Kee-Kahb
    • Applied Chemistry for Engineering
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    • v.26 no.6
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    • pp.730-736
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    • 2015
  • Cyclotrimethylene trinitramine (RDX) is a high explosive commonly used for military applications. Submicronization of RDX particles has been a critical issue in order to alleviate the unintended and accidental stimuli toward safer and more powerful performances. The purpose of this study is to optimize experimental variables for drowning-out crystallization applied to produce submicron RDX particles. Effects of RDX concentration, anti-solvent temperature and anti-solvent mass were analyzed by the central composite rotatable design. The adjusted determination coefficient of regression model was calculated to be 0.9984 having the p-value less than 0.01. Response surface plots based on the central composite rotatable design determined the optimum conditions such as RDX concentration of 3 wt%, anti-solvent temperature of $0.2^{\circ}C$ and anti-solvent mass of 266 g. The optimum and experimental diameters of RDX particles were measured to be $0.53{\mu}m$ and $0.53{\mu}m$, respectively. The regression model satisfactorily predicts the average diameter of RDX particles prepared by drowning-out crystallization. Structure of RDX crystals was found to be ${\alpha}$-form by X-ray diffraction analysis and FT-IR spectroscopy.

Changes in CGRP-immunoreactive Nerve Fibers during Expansion of Midpalatal Suture of the Rat (백서 정중구개봉합 확대후의 CGRP 면역반응 신경섬유의 변화)

  • Kim, Bo-Kyung;Park, Kuk-Pil;Kyung, Hee-Moon;Kwon, Oh-Won;Sung, Jae-Hyun
    • The korean journal of orthodontics
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    • v.29 no.1 s.72
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    • pp.73-81
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    • 1999
  • Midpalatal suture expansion is often used for patients haying narrow maxillary arch, cleft palate, respiratory handicap with narrow nasal cavity. CGRP has been known as a modulator of pain transmission in central nervous system and a local effector to peripheral tissue causing vasodilation, increase of blood flow, modulation of immune system, regulation of macrophagic function and stimulation of bone formation. To investigate changes of CGRP-immunoreactive nerve fibers in midpalatal suture during the expansion, immunohistochemical study was performed by using rats. Experimental rats (10 weeks, 250 gm) were divided into five groups (control, 1, 4, 7, 14 days group (each n=4) and applied orthodontic force (approximately 200gm) to upper anterior incisors. Frozen sections of midpalatal suture area were immunostained by using rabbit antisera. The results were as follows. ${\cdot}$ The CGRP-immunoreactive nerve fibers were hardly observed in control group. ${\cdot}$ In 1 day group, the CGRP-immunoreactive nerve fibers were more increased around the vessels than control group. ${\cdot}$ In 4 days group, the CGRP-immunoreactive nerve fibers were more increased than control group, but not more increased than 1 day group. Vascular diameter was more enlarged. ${\cdot}$ In 7 days group, especially, hematoxilin affinity of cells was remarkable and cells were arranged along the bone margin. The CGRP-immunoreactive nerve fibers were more reduced than 4 days group and vascular diameter was also reduced. ${\cdot}$ In 14 days group, the CGRP-immunoreactive nerve fibers were similar to those of 7 days group and the irregularity of bone margin was almost recoverd. In Conclusion, the CGRP-immunoreactive nerve fibers nay be related to initial neurogenic inflammatory reaction in expanding mid-palatal suture.

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Expression of Growth Factors and Secretory Leukocyte Protease Inhibitor (SLPI) in RAW264.7 Cells after Lipopolysaccharide (LPS) Stimulation (대식세포주 (RAW264.7)에서 박테리아성 지질다당류 (LPS) 자극에 의한 분비백혈구단백분해효소억제제 (SLPI)와 성장인자들의 발현)

  • Son, Wook-Hee;Choi, Baik-Dong;Jeong, Soon-Jeong;Wang, Guan-Lin;Hwang, Ho-Keel;Jeong, Moon-Jin
    • Applied Microscopy
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    • v.37 no.2
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    • pp.93-102
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    • 2007
  • Secretory leukocyte protease inhibitor (SLPI) was known as one of bacterial lipopolysaccharide (LPS)-induced products of macrophage. Macrophages play an important role in the development of inflammatory responses by secreting an array of cytokines and chemokines in a tissue microenvironment. To identify the function and relationship between potent growth factors and SLPI after LPS stimulation, we conducted reverse transcriptase polymerase chain reaction (RT-PCR) and Western blots for the detection of mRNA and protein expression of SLPI and growth factors such as VEGF, PDGF, bFGF after 100 ng LPS stimulation on the RAW264.7 cells. The result of RT-PCR was showed SLPI mRNA expression was increased from 60 min to 48h in RAW 264.7 cells after incubation with LPS. VEGF and PDGF mRNA was expressed highly at initial stage by LPS stimulation. The mRNA of bFGF and type I collagen was very weakly expressed after LPS stimulation. SLPI protein level was increased likely the mRNA levels in RAW 267.7 cells. Additionally, phase contrast and scanning electron microscopic observation demonstrated that the LPS induce the change of morphology of the RAW264.7 cells. From these results, it suggest that expression of SLPI by LPS treatment may associate with VEGF and PDGF expression in RAW264.7 cells.

Effect of Repetitive Transcranial Magnetic Stimulation in Drug Resistant Depressed Patients (치료 저항성 우울증 환자에서 반복적 경두개 자기자극후 국소뇌혈류 변화)

  • Chung, Yong-An;Yoo, Ie-Ryung;Kang, Bong-Joo;Chae, Jeong-Ho;Lee, Hye-Won;Moon, Hyun-Jin;Kim, Sung-Hoon;Sohn, Hyung-Sun;Chung, Soo-Kyo
    • Nuclear Medicine and Molecular Imaging
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    • v.41 no.1
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    • pp.9-15
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    • 2007
  • Purpose: Repetitive transcranial magnetic stimulation (rTMS) has recently been clinically applied in the treatment of drug resistant depressed patients. There are mixed findings about the efficacy of rTMS on depression. Furthermore, the influence of rTMS on the physiology of the brain is not clear. We prospectively evaluated changes of regional cerebral blood flow (rCBF) between pre- and post-rTMS treatment in patients with drug resistant depression. Materials and Methods: Twelve patients with drug-resistant depression (7 male, 5 female; age range: $19{\sim}52$ years; mean age: $29.3{\pm}9.3$ years) were given rTMS on right prefrontal lobe with low frequency (1 Hz) and on left prefrontal lobe with high frequency (20 Hz), with 20-minute-duration each day for 3 weeks. Tc-99m ECD brain perfusion SPECT was obtained before and after rTMS treatment. The changes of cerebral perfusion were analyzed using statistical parametric mapping (SPM; t=3.14, uncorrected p<0.01, voxel=100). Results: Following areas showed significant increase in rCBF after 3 weeks rTMS treatment: the cingulate gyrus, fusiform gyrus of right temporal lobe, precuneus, and left lateral globus pallidus. Significant decrement was noted in: the precental and middle frontal gyrus of right frontal lobe, and fusiform gyrus of left occipital lobe. Conclusion: Low-frequency rTMS on the right prefrontal cortex and high-frequency rTMS on the left prefrontal cortex for 3 weeks as an add-on regimen have increased and decreased rCBF in the specific brain regions in drug-resistant depressed patients. Further analyses correlating clinical characteristics and treatment paradigm with functional imaging data may be helpful in clarifying the pathophysiology of drug-resistant depressed patients.

Waveform Sorting of Rabbit Retinal Ganglion Cell Activity Recorded with Multielectrode Array (다채널전극으로 기록한 토끼 망막신경절세포의 활동전위 파형 구분)

  • Jin Gye Hwan;Lee Tae Soo;Goo Yang Sook
    • Progress in Medical Physics
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    • v.16 no.3
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    • pp.148-154
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    • 2005
  • Since the output of retina for visual stimulus is carried by neurons of very diverse functional properties, it is not adequate to use conventional single electrode for recording the retinal action potential. For this purpose, we used newly developed multichannel recording system for monitoring the simultaneous electrical activities of many neurons in a functioning piece of retina. Retinal action potentials are recorded with an extra-cellular planar array of 60 microelectrodes. In studying the collective activity of the ganglion cell population it is essential to recognize basic functional distinctions between individual neurons. Therefore, it is necessary to detect and to classify the action potential of each ganglion cell out of mixed signal. We programmed M-files with MATLAB for this sorting process. This processing is mandatory for further analysis, e.g. poststimulus time histogram (PSTH), auto-correlogram, and cross-correlogram. We established MATLAB based protocol for waveform classification and verified that this approach was effective as an initial spike sorting method.

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