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Anti-inflammatory effect of Sinhyowoldo-san Extract with regard to Pro-inflammatory Mediators in PMA plus A23187-induced Human Mast Cells (인간 비만세포에서 PMA와 A23187에 의해 유도된 전염증 매개체에 대한 신효월도산 추출물의 항염증 효과)

  • Wi, Gyeong;Yang, Da-Wun;Kang, Ok-Hwa;Kim, Sung-Bae;Mun, Su-Hyun;Seo, Yun-Soo;Kang, Da-Hye;Lim, Jae-Soo;Kim, Ma-Ryong;Kwak, Nam-Won;Kong, Ryong;Kwon, Dong-Yeul
    • The Korea Journal of Herbology
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    • v.29 no.6
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    • pp.117-123
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    • 2014
  • Objectives : Sinhyowoldo-san (SHWDS) is said to be a traditional medicine used for shigellosis, abdominal pain, diarrhea. But mechanism of SHWDS mediated-modulation of immune function is not sufficiently understood. To ascertain the molecular mechanisms of SHWDS 70% EtOH extract on pharmacological and biochemical actions in inflammation, we researched the effect of pro-inflammatory mediators in phorbol-12-myristate-13-acetate (PMA)+ A23187-activated human mast cell line (HMC-1). Methods : In the present research, cell viability was measured by MTS assay. pro-inflammatory cytokine production was measured by performing enzyme-linked immunosorbent assay (ELISA), reverse transcription polymerase chain reaction (RT-PCR), and western blot analysis to analyze the activation of mitogen-activated protein kinases (MAPKs), nuclear factor kappa-light-chain-enhancer of activated B cells ($NF-{\kappa}B$). The investigation focused on whether SHWDS inhibited the expressions of interleukin-6 (IL-6), interleukin-8 (IL-8), MAPKs and $NF-{\kappa}B$ in PMA+A23187-activated HMC-1 cells. Results : SHWDS has no cytotoxicity at measured concentration (50, 100, and $250{\mu}g/ml$). SHWDS ($250{\mu}g/ml$) inhibits pro-inflammatory cytokine expression in PMA+ A23187-activated HMC-1 cells. Moreover, SHWDS inhibited cyclooxygenase (COX)-2 expression. In activated HMC-1 cells, SHWDS suppressed phosphorylation of extracellular signal-regulated kinase (ERK 1/2) and c-jun N-terminal Kinase (JNK 1/2). Then, SHWDS suppressed activation of nuclear factor $NF-{\kappa}B$ in nuclear, degradation of IkB ${\alpha}$ in cytoplasm. Conclusions : We propose that SHWDS has an anti-inflammatory therapeutic potential, which may result from inhibition of ERK 1/2, JNK 1/2 phosphorylation and $NF-{\kappa}B$ activation, thereby decreasing the expression of pro-inflammatory genes.

Effects of Bisphenol A on the Placental Function and Reproduction in Rats (Bisphenol A가 흰쥐의 태반 기능과 출산에 미치는 영향)

  • Lee, Chae-Kwan;Kim, Seog-Hyun;Moon, Deog-Hwan;Kim, Jeong-Ho;Son, Byung-Chul;Kim, Dae-Hwan;Lee, Chang-Hee;Kim, Hwi-Dong;Kim, Jung-Won;Kim, Jong-Eun;Lee, Chae-Un
    • Journal of Preventive Medicine and Public Health
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    • v.38 no.3
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    • pp.330-336
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    • 2005
  • Objectives : The aim of this study was to investigate the effects of bisphenol A (BPA), an estrogen-like environmental endocrine disrupter, on the placental function and reproduction in rats. The mRNA levels of the placental prolactin-growth hormone(PRL-GH) gene family, placental trophoblast cell frequency and reproductive data were analyzed. Methods : The pregnancies of F344 Fisher rats ($160g{\pm}20g$) were detected by the presence of the copulatory plug or sperm in the vaginal smear, which marked Day 0 of pregnancy. Pregnant rats were divided into three groups. The control group was intraperitoneally injected with a sesame oil vehicle. The two remaining groups were injected with 50 or 500 mg/kg B.W/day of BPA, resuspended in sesame oil, on either days 7 to 11 or 16 to 20 of pregnancy, with the rats sacrificed on either day 11 or 20, respectively. The mRNA levels of PRL-GH and Pit-1a and b isotype genes were analyzed by Northern blot hybridization and reverse transcription-polymerase chain reaction. The hormone concentrations were analyzed by radioimmunoassay, and the frequency of the placental trophoblast cells observed by a histochemical study. Reproductive data, such as the placental weight and litter size, were surveyed on day 20. The fetal weight was surveyed for 4 weeks after birth. A statistical analysis was carried out using the SAS program (version 8.1). Results : The mRNA levels of the PRL-GH gene family, such as placental lactogen I, Iv and II, prolactin like protein A, C and Cv, and decidual prolactin-related protein were significantly reduced due to BPA exposure. The mRNA levels of the Pit-1a and b isotype genes, which induce the expression of the PRL-GH gene family in the rat placenta, were also reduced due to BPA exposure. The PL-Iv and PL-II concentrations were reduced in the BPA exposed group. During the middle to last stage of pregnancy (Days 11-20), a high dose of BPA exposure reduced the frequency of spongiotrophoblast cells, which are responsible for the secretion of the PRL-GH hormones. Reproductive data, such as the placental and fetal weights and the litter size, were reduced, but that of the pregnancy period was extended in the BPA exposed compared to the control group. Conclusions : BPA disrupts the placental functions in rats, which leads to reproductive disorders.

The Change of Cell-cycle Related Proteins and Tumor Suppressive Effect in Non-small Cell Lung Cancer Cell Line after Transfection of p16(MTS1) Gene (폐암세포에 p16 (MTS1) 유전자 주입후 암생성능의 변화 및 세포주기관련 단백질의 변동에 관한 연구)

  • Kim, Young-Whan;Kim, Jae-Yeol;Yoo, Chul-Gyu;Han, Sung-Koo;Shim, Young-Soo;Lee, Kye-Young
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.4
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    • pp.796-805
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    • 1997
  • Background : It is clear that deregulation of cell cycle progression is a hallmark of neoplastic transformation and genes involved in the $G_1$/S transition of the cell cycle are especially frequent targets for mutations in human cancers, including lung cancer. p16 gene product, one of the G1 cell-cycle related proteins, that is recently identified plays an important role in the negative regulation of the the kinase activity of the cyclin dependent kinase (cdk) enzymes. Therefore p16 gene is known to be an important tumor suppressor gene and is also called MTS1 (multiple tumor suppressor 1). No more oncogenes have been reported to be frequently related to multiple different malignancies than the alterations of p16 gene. Especially when it comes to non-small cell lung cancer, there was no expression of p16 in more than 70% of cell lines examined. And also it is speculated that p16 gene could exert a key role in the development of non-small cell lung cancer. This study was designed to evaluate whether p16 gene could be used as a candidate for gene therapy of non-small cell lung cancer. Methods : After the extraction of total RNA from normal fibroblast cell line and subsequent reverse transcriptase reaction and polymerase chain reaction, the amplified p16 cDNA was subcloned into eukaryotic expression plasmid vector, pRC-CMV. The constructed pRC-CMV-p16 was transfected into the NCI-H441 NSCLC cell line using lipofectin. The changes of G1 cell-cycle related proteins were investigated with Western blot analysis and immunoprecipitation after extraction of proteins from cell lysates and tumor suppressive effect was observed by clonogenic assay. Results : (1) p16(-) NCI-H441 cell line transfected with pRC-CMV-p16 showed the formation of p16 : cdk 4 complex and decreased phosphorylated Rb protein, while control cell line did not. (2) Clonogenic assay demonstrated that the number of colony formation was markedly decreased in p16(-) NCI-H441 cell line transfected with pRC-CMV-p16 than the control cell line. Conclusion : It is confirmed that the expression of p16 protein in p16 absent NSCLC cell line with the gene transfection leads to p16 : cdk4 complex formation, subsequent decrease of phosphorylated pRb protein and ultimately tumor suppressive effects. And also it provides the foundation for the application of p16 gene as a important candidate for the gene therapy of NSCLC.

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Expression of Serotonin(5-HT) Receptor Isotypes in Reproductive Organs of Male Rat (수컷 흰쥐 생식기관에서의 세로토닌 수용체 아형 유전자 발현)

  • 이성호
    • Development and Reproduction
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    • v.6 no.2
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    • pp.111-115
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    • 2002
  • 5-Hydroxytryptamine(5-HT; serotonin) system has been implicated in the modulation of male sexual behaviors and the secretion of reproductive hormones. In human males, selective serotonin re-uptake inhibitors(SSRIs) are known to improve the major male sexual dysfunction, premature ejaculation, through the central nervous system-mediated pathways. As numerous hormone and local factors, 5-HT may have peripheral role in the regulation of male sexual function. The expression of 5-HT receptor subtypes in the target tissue, however, has not been explored yet. The present study was undertaken to test whether the 5-HT receptor subtypes are expressed in the reproductive tissues of male rat, especially in ejaculatory machinery such as seminal vesicle and vas deferens. To do this, reverse transcription-polymerase chain reaction(RT-PCR) and Southern blot analysis were employed. The transcripts for the 1A, 1B and 2C subtypes of 5-HT receptor were amplified in all the tested tissues. The present study demonstrated the expression of 5-HT receptor in the rat ejaculatory machinery, suggesting that 5-HT may play a pivotal role in the male sexual function via not only central pathway but also peripheral route. Further study on the receptor subtype-specific effect and their harmonized mode of action will be needed to establish the understanding of ejaculation mechanism and drug design.

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Toll-like Receptor 4-mediated Apoptotic Cell Death in Primary Isolated Human Cervical Cancers (부인과질환 특이적 종양의 TLR4 매개성 apoptosis 유발에 관한 연구)

  • Won, Jinyoung;Hong, Yunkyung;Park, Sookyoung;Kim, Joo-Heon;Hong, Yonggeun
    • Journal of Life Science
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    • v.28 no.6
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    • pp.718-725
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    • 2018
  • Toll-like receptor 4 (TLR4) has been implicated in cell proliferation and apoptosis in several types of cancer. In this study, the impact of TLR4 activation on apoptotic cell death in gynecologic cancers induced by lipopolysaccharide (LPS) was investigated. Cervical cancer cell lines were produced from isolated surgical specimens supplied by Paik Hospital. The primary cultures of normal myometrium and gynecologic cancers, including cervical, endometrial, and ovarian cancers, were used to examine the differences in morphological characteristics between normal and cancerous cells. A reverse transcription polymerase chain reaction analysis was used to determine the relative expression levels of TLR4 gene involved in apoptosis-associated signaling in cervical cancer cells. The cancer cell colonies showed a tendency to reach high levels of confluency compared with normal cells. In addition, an enhanced growth rate and loss of contact inhibition were observed in gynecologic cancer cells compared with normal cells (doubling times of 16.6 hr vs. 26 hr, respectively). The expression level of ITGA5, an alpha-5 integrin marker, was upregulated in normal myometrial cells, but this tendency was not exhibited in cervical cancer cells. Furthermore, p53 tumor suppressor gene expression was upregulated, whereas TLR4 and caspase-3 gene expressions were downregulated in cervical cancer cells. Notably, the expression levels of TLR4 and caspase-3 were increased significantly in LPS-treated cancer cells compared with those in non-LPS-treated cells. These results suggest that the TLR4-mediated caspase-dependent apoptotic signaling pathway could be suggested as a therapeutic target for the treatment of gynecologic cancers, including cervical cancers.

Studies on Simultaneous Determination of Chlorophyll a and b, Pheophorbide a, and $\beta-Carotene$ in Chlorella and Spirulina Products (클로렐라 및 스피루리나제품에 함유된 엽록소 a, b, 페오포르바이드 a 및 $\beta$-카로틴의 동시분석법에 관한 연구)

  • Lee Young Ja;Kim So Hee;Kim Jin-Sook;Han Jeong A;Seo Hae Jeom;Lim Hyo Jeong;Choi Soo Young
    • Journal of Food Hygiene and Safety
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    • v.20 no.3
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    • pp.141-146
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    • 2005
  • A simple and sensitive analysis method based on reverse phase (RP) HPLC with UV detector was developed for simultaneous determination of chlorophyll a and b, pheophorbide a and $\beta-Carotene$ in Chlorella and Spirulina products. For added concentration $(50\;\mug/ml)$ of chlorophyll a and b, pheophorbide a and $\beta-Carotene$, recoveries of those were 70.3, 71.6, 60.1 and $90.5\%$, respectively, with relative standard deviations of 2.8,6.0, 10.6 and $10.4\%$. Limit of detection and quantification had ranges of $0.1\sim1.0\;\mug/ml$ and $0.2\sim2.0\;\mug/ml$, respectively. Calibration curve was linear with correlation coefficient of 0.995 for chlorophyll a and b, pheophorbide a and $\beta-Carotene$. Results of simultaneous determination in Chlorella and Spirulina products were showed ranges of $121.g\sim543.0\;\mug/ml$ for chlorophyll a,$0.6\sim160.0\;\mug/ml$ for chlorophyll b, $19.2\sim60.3\;\mug/ml$ for pheophorbide a and $383.6\sim1713.7\;\mug/ml$ for $\beta-Carotene$, respectively. Chlorophyll b contents in Chlorella products were detected above 30 times level to those in Spirulina products. $\beta-Carotene$ contents in Spirulina products were detected 2.7 times level to those in Chlorella products.

Identification of a New Potyvirus, Keunjorong mosaic virus in Cynanchum wilfordii and C. auriculatum (큰조롱과 넓은잎 큰조롱에서 신종 포티바이러스(큰조롱모자이크바이러스)의 동정)

  • Lee, Joo-Hee;Park, Seok-Jin;Nam, Moon;Kim, Min-Ja;Lee, Jae-Bong;Sohn, Hyoung-Rac;Choi, Hong-Soo;Kim, Jeong-Soo;Lee, Jun-Seong;Moon, Jae-Sun;Lee, Su-Heon
    • Research in Plant Disease
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    • v.16 no.3
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    • pp.238-246
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    • 2010
  • In 2006 fall, a preliminary survey of viruses in two important medicinal plants, Cynanchum wilfordii and C. auriculatum, was conducted on the experimental fields at the Agricultural Research and Extension Services of Chungbuk province in Korea. On each experimental fields, percentage of virus infection was ranged from 20 to 80%, and especially an average of disease incidence propagated by roots was twice higher than that by seeds. The various symptoms were observed in Cynanchum spp. plants, such as mosaic, mottle, necrosis, yellowing, chlorotic spot and malformation etc. In electron microscopic examination of crude sap extracts, filamentous rod particles with 390-730 nm were observed in most samples. The virus particles were purified from the leaves of C. wilfordii with typical mosaic symptom, and the viral RNA was extracted from this sample containing 430-845 nm long filamentous rod. To identify the viruses, reverse transcription followed by PCR with random primers was carried out. The putative sequences of P3 and coat protein of potyvirus were obtained. From a BLAST of the two sequences, they showed 26-38% and 62-72% identities to potyviruses, respectively. In SDS-PAGE analysis, the subunit of coat protein was approximately 30.3 kDa, close to the coat protein of potyvirus. In bioassay with 21 species in 7 families, Chenopodium quinoa showed local lesion on inoculated leave and chlorotic spot on upper leave, but the others were not infected. RT-PCR detection using specific primer of C. wilfordii and C. auriculatum samples, all of 24 samples with virus symptom was positive, and five out of seven samples without virus symptom were also positive. On the basis of these data, the virus could be considered as a new member of potyvirus. We suggested that the name of the virus was Keunjorong mosaic virus (KjMV) after the common Korean name of C. wilfordii.

Effects of Chromium (VI) Exposure on the Placental Function and Reproduction in Rats (6가 크롬 폭로가 랫트의 태반 기능과 출산에 미치는 영향)

  • Lee, Heun;Moon, Deog-Hwan;Lee, Chae-Un;Kang, Sung-Goo;Son, Byung-Chul;Kim, Dae-Hwan;Lee, Chang-Hee;Kim, Jung-Won;Lee, Chae-Kwan;Chun, Jin-Ho
    • Journal of Preventive Medicine and Public Health
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    • v.37 no.2
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    • pp.157-165
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    • 2004
  • Objectives : This study aimed to investigate the toxic effects of chromium (VI) on the placental function and reproduction in rats. For the study, the placental prolactin-growth hormone (PRL-GH) gene expression, placental trophoblast cell differentiation and reproductive data were analyzed. Methods : The pregnancies of F344 Fisher rats were checked by the presence of a copulatory plug or sperm in the vaginal smear, which was defined as day 0 of the pregnancy. Pregnant rats were divided into the three groups. The control group was given tap water (chromium level < 0.001 ppm) and the remaining groups were given 250 or 750 ppm of chromium (VI) [as potassium dichromate], from day 7 to 19 of the pregnancy. Rats were sacrificed at days 11 and 20 of pregnancy. The mRNA levels of PRL-GH and Pit-1a and b isotype genes were analyzed by Northern blot hybridization and reverse transcription-polymerase chain reaction (RT-PCR). The hormonal concentration was analyzed by radioimmunoassay, and the differentiation of placental trophoblast cells were observed by histochemical studies. Reproductive data, such as placental and fetal weights, pregnancy period, and litter size, were surveyed at day 20 of pregnancy and after birth. A statistical analysis was carried out using the SAS program (version 8.1). Results : The mRNA levels of the prolactin-growth hormone (PRL-GH) family of genes were dose dependently reduced by chromium exposure. The mRNA levels of Pit-1a and b isotype genes that induce the expression of the PRL-GH family of genes were also reduced by chromium exposure. The PRL-GH hormonal concentration in the rat placenta, fetus and maternal blood were decreased by chromium exposure. In the middle stage of pregnancy (day 11), a high dose of chromium suppressed the differentiation of spongiotrophoblast cells that secret the PRLGH hormones. In the last stage of pregnancy (day 20), a high dose of chromium induced apoptosis of placental cells. Reproductive data, such as placental and fetal weights, litter size, were reduced, but the pregnancy period was extended in the group exposed to chromium compared with the controls. Conclusion : Chromium (VI) disrupts the ordered functions of the placenta, which leads to reproductive disorders in rats.

Analysis on Practicality of Seed Treatments for Medicinal Plants Published in Korean Scientific Journals (국내 학술지에 발표된 약용작물 종자처리의 실용성 분석)

  • Kang, Jin-Ho;Yoon, Soo-Young;Jeon, Seung-Ho
    • Korean Journal of Medicinal Crop Science
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    • v.12 no.4
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    • pp.328-341
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    • 2004
  • Presowing seed treatments used to enhance the rates of germination and afterward seedling emergence have not occasionally shown the same rate in indoor and field. The treatments considering germination mechanism and factors affecting germination must be totally included in indoor experiments so that the results drawn can be reproduced in the field. Seed germination is controlled by Phytochrome-mediated action changed with composition rates of red and far-red lights. Sunlight can penetrate soil into $6{\sim}9\;mm$ depth, which in turn means that seeds having $2{\sim}3\;mm$ in their width may receive the light if soil was covered 3 times over them. The penetrating light, moreover, turns to more far-red light than red light reverse to the sunlight. For germination tests after the artificial presowing seed treatments, therefore, seeds of smaller than 2 mm (< 2 mm), $2{\sim}3\;mm$, and larger than 3 mm (> 3 mm) must be done with incandescent lamp (IL) having more far-red light, with IL or in darkness, and in darkness, respectively. The 96 papers published in 13 Korean scientific journals up to the end of 2003 were analysed on the basis of the above explanation. 91 species were used 147 times as experimental materials; 101 times for < 2 mm seeds, 24 times for $2{\sim}3\;mm$ seeds and 22 times for > 3 mm seeds. If they were analysed as the light sources used for germination tests, correct applications reached more and less than 60% in both $2{\sim}3\;mm$ and > 3 mm seeds but 23% in < 2 mm seeds, conclusionally meaning that when the experimental results in the scientific papers were applied into farming practices, care was taken of their application because most of medicinal plant seeds were very small.

A development of DS/CDMA MODEM architecture and its implementation (DS/CDMA 모뎀 구조와 ASIC Chip Set 개발)

  • 김제우;박종현;김석중;심복태;이홍직
    • The Journal of Korean Institute of Communications and Information Sciences
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    • v.22 no.6
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    • pp.1210-1230
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    • 1997
  • In this paper, we suggest an architecture of DS/CDMA tranceiver composed of one pilot channel used as reference and multiple traffic channels. The pilot channel-an unmodulated PN code-is used as the reference signal for synchronization of PN code and data demondulation. The coherent demodulation architecture is also exploited for the reverse link as well as for the forward link. Here are the characteristics of the suggested DS/CDMA system. First, we suggest an interlaced quadrature spreading(IQS) method. In this method, the PN coe for I-phase 1st channel is used for Q-phase 2nd channels and the PN code for Q-phase 1st channel is used for I-phase 2nd channel, and so on-which is quite different from the eisting spreading schemes of DS/CDMA systems, such as IS-95 digital CDMA cellular or W-CDMA for PCS. By doing IQS spreading, we can drastically reduce the zero crossing rate of the RF signals. Second, we introduce an adaptive threshold setting for the synchronization of PN code, an initial acquistion method that uses a single PN code generator and reduces the acquistion time by a half compared the existing ones, and exploit the state machines to reduce the reacquistion time Third, various kinds of functions, such as automatic frequency control(AFC), automatic level control(ALC), bit-error-rate(BER) estimator, and spectral shaping for reducing the adjacent channel interference, are introduced to improve the system performance. Fourth, we designed and implemented the DS/CDMA MODEM to be used for variable transmission rate applications-from 16Kbps to 1.024Mbps. We developed and confirmed the DS/CDMA MODEM architecture through mathematical analysis and various kind of simulations. The ASIC design was done using VHDL coding and synthesis. To cope with several different kinds of applications, we developed transmitter and receiver ASICs separately. While a single transmitter or receiver ASC contains three channels (one for the pilot and the others for the traffic channels), by combining several transmitter ASICs, we can expand the number of channels up to 64. The ASICs are now under use for implementing a line-of-sight (LOS) radio equipment.

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