• Title/Summary/Keyword: adjuvants

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Production of polyclonal anti-$\beta$-adrenergic receptor antibody and it′s effects on receptor ligand binding

  • Kim, Hee-Jin;Shin, Chan-Young;Noh, Min-Su;Ko, Kwang-Ho
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1995.04a
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    • pp.86-86
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    • 1995
  • The analysis of membrane receptors for hormones and neurotransmitters has progressed considerably by pharmacological and biochemical means and more recently by the use of specific anti-receptor antibodies. A 14-mer peptide (from Phe102 to Leu115 of ${\beta}$2-adrenergic receptor) was synthesized and this peptide was coupled to carrier protein Keyhole Limpet Hemocyanin(KLH) by glutaraldehyde method. A 0.5mg of KLH-coupled peptide was emulsified with equal volume of complete Freund's adjuvant and injected via popliteal lymph node to each of the three Newzealnd White rabbits. Booster injections were repeated at 4 weeks interval for three times with incomplete Freund's adjuvants. One week after the final injection, serum was prepared from ear artery. Nonspecific immunoglobulins were removed by passing the serum through KLH-Sepharose 6B affinity matrix and further by incubation with bovine lung aceton powder. The titer of the antibody for synthetic peptide which was determined by enzyme linked immunosorbent assay(ELISA) was about l/l,000. The antibody produced in this study revealed 67kDa protein band in the western blot of partially purified guinea pig lung ${\beta}$-adrenergic receptor preparation. The antibody inhibited ${\beta}$-adrenergic antaginist [3H] Dihydroalprenolol binding to soluble ${\beta}$-adrenergic receptor by 25% while control sera did not show any inhibitory effects, The result of this study suggests that the peptide sequence selected in this study may play some important roles in adrenergic receptor-ligand interaction.

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Enzyme-Linked Immunosorbent Assay for Detection of Nivalenol (Nivaleno의 검출을 위한 효소 면역 측정법)

  • 손동화;이향범;곽보연;김수호;권창희
    • Journal of Food Hygiene and Safety
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    • v.13 no.2
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    • pp.129-134
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    • 1998
  • To develop an enzyme-linked immunosorbent assay (ELISA) for nivalenol (NIV), we produced polyclonal antibodies against tetraacetyl nivalenol (Ac4-NIV) and established ELISA conditions. Ac4-NIV-hemisuccinate conjugated to bovine serum albumin (Ac4-NIV-HS-BSA) was immunized with Freund's adjuvants into rabbits subcutaneously several times. By use of the antiserum showing the highest titer and Ac4-NIV-HS-HRP conjugate, we established competitive direct ELISA (cdELISA). Standard curve of cdELISA showed that the detection range of Ac4-NIV was about 10~5,000 ng/ml (ppb). The cross-reactivities of the polyclonal antibody towards Ac4-NIV and acetyl T-2 were 100 and 70% respectively, and those towards NIV, deoxynivalenol, 3-acetyl deoxynivalenol, 15-acetyl deoxynivalenol, triacetyl deoxynivalenol, fusarenonX, and T-2 were less than 0.1%. When cdELISA was applied to NIV-spiked corns followed by extraction with 70% acetonitrile and acetylation with acetic anhydride in pyridine, the recovery rates of the Ac4-NIV were 108, 143, and 70% (average, 107%) in the levels of 100, 300, and 1,000 ng/g (ppb), respectively.

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Cytokine Reporter Mouse System for Screening Novel IL12/23 p40-inducing Compounds

  • Im, Wooseok;Kim, Hyojeong;Yun, Daesun;Seo, Sung-Yum;Park, Se-Ho;Locksley, Richard M.;Hong, Seokmann
    • Molecules and Cells
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    • v.20 no.2
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    • pp.288-296
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    • 2005
  • Cytokines interleukin (IL) 12 and 23 play critical roles in linking innate and adaptive immune responses. They are members of heterodimeric cytokines, sharing a subunit p40. Although IL12/23 p40 is mainly induced in macrophages and dendritic cells (DCs) after stimulation with microbial Toll-like receptor ligands, methods to monitor the cells that produce IL12/23 p40 in vivo are limited. Recently, the mouse model to track p40-expressing cells with fluorescent reporter, yellow fluorescent protein, has been developed. Macrophages and DCs from these mice faithfully reported p40 induction using the fluorescent marker. Here we took advantage of these reporter mice to screen bio-compounds for p40-inducing activity. After screening hundreds of compounds, we found several extracts inducing IL12/23 p40 gene expression. Treatment of DCs with these extracts induced the expression of MHC class II and co-stimulatory molecules, which implies that these might be useful as adjuvants. Next, the in vivo target immune cells of candidate compounds were examined. The reporter system can be useful to identify cells producing IL12 or IL23 in vivo as well as in vitro. Thus, our cytokine reporter system proved to be a valuable reagent for screening for immunostimulatory molecules and identification of target cells in vivo.

Optimization and High-level Expression of a Functional GST-tagged rHLT-B in Escherichia coli and GM1 Binding Ability of Purified rHLT-B

  • Ma Xingyuan;Zheng Wenyun;Wang Tianwen;Wei Dongzhi;Ma Yushu
    • Journal of Microbiology
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    • v.44 no.3
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    • pp.293-300
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    • 2006
  • The Escherichia coli heat-labile enterotoxin B subunit (HLT-B) is one of the most powerful mucosal immunogens and known mucosal adjuvants. However, the induction of high levels of HLT-B expression in E. coli has proven a difficult proposition. Therefore, in this study, the HLT-B gene was cloned from pathogenic E. coli and expressed as a fusion protein with GST (glutathion S-transferase) in E. coli BL2l (DE3), in an attempt to harvest a large quantity of soluble HLT-B. The culture conditions, including the culture media used, temperature, pH and the presence of lactose as an inducer, were all optimized in order to obtain an increase in the expression of soluble GST-rHLT-B. The biological activity of the purified rHLT-B was assayed in a series of GMI-ELISA experiments. The findings of these trials indicated that the yield of soluble recombinant GST-rHLT-B could be increased by up to 3-fold, as compared with that seen prior to the optimization, and that lactose was a more efficient alternative inducer than IPTG. The production of rHLT-B, at 92 % purity, reached an optimal level of 96 mg/l in a 3.7 L fermentor. The specific GM1 binding ability of the purified rHLT-B was determined to be almost identical to that of standard CTB.

Post-operative analgesia of 2% lignocaine with or without magnesium sulfate for inferior alveolar nerve block in symptomatic mandibular molars - a randomized double blind controlled clinical trial

  • Chandrasekaran, Charanya;Vijay, Amirtharaj L;Sekar, Mahalaxmi;Mary, Nancy S
    • Journal of Dental Anesthesia and Pain Medicine
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    • v.20 no.3
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    • pp.147-154
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    • 2020
  • Background: Single inferior alveolar nerve block is ineffective in achieving adequate pulpal anesthesia in 30-80% of patients due to anatomical variations, local tissue pH, central sensitization, and several factors. Various supplementary techniques and combination of adjuvants with lignocaine are used to overcome these failures. Magnesium sulfate (MgSO4), one such adjuvant, acts at the N-methyl-D-aspartate glutamate receptor resulting in effective anesthesia. The aim of this prospective, randomized, double-blind, clinical controlled trial was to evaluate the onset, anesthetic efficacy, duration and post-operative analgesia of 2% lignocaine with and without the addition of MgSO4 in patients with symptomatic irreversible pulpitis and apical periodontitis. Methods: Fourty-two patients were randomly divided into three groups: 2% lignocaine (group 1) and 2% lignocaine with MgSO4 (75 mg) and (150 mg) in groups 2 and 3, respectively. Pre-operative vitals and Heft Parker-Visual Analogue Scale (HP-VAS) pain scores were recorded. The onset of anesthesia, anesthetic efficacy, and duration of anesthesia were evaluated post administration of the local anesthetic solution. The post-operative analgesia was examined at intervals of 2, 6, 12, 24, and 48 h. Results: Administration of 150 mg MgSO4 hastens the onset of anesthesia (1.29 min) and produces better anesthetic efficacy (3.29 HP-VAS) compared to group 2 (2.07 min and 9.14 HP-VAS) and group 1 (3.29 min and 35.79 HP-VAS), respectively. The duration of anesthesia was significantly higher in group 3 (247.07 min) compared to that of groups 2 and 1 (190 min and 110.21 min) with P < 0.05. Conclusion: Combining 75 mg or 150 mg of MgSO4 with lignocaine is more effective than 2% lignocaine and 75 mg of MgSO4 is adequate for endodontic procedures.

The immune-adjuvant effect and safety of recombinant CC chemokine 1 (rRbCC1) in rock bream, Oplegnathus fasciatus

  • Kwon, Mun-Gyeong;Kim, Ju-Won;Hwang, Seong-Don;Kim, Eun-Gyeong;Park, Dae-Won;Park, Chan-Il
    • Journal of fish pathology
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    • v.26 no.3
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    • pp.231-240
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    • 2013
  • Adjuvants are immune enhancers that are often used in vaccination to augment the immune response of a vaccine, thereby enhancing the protective immunity against the targeted disease. In the present study, we used the recombinant protein, such as rRbCC1, this protein was produced from rock bream CC chemokine 1. To verify the adjuvant effects of this recombinant protein, the immune responses of rock bream to Streptococcus iniae (S. iniae) FKC vaccination, which alone or in combination with recombinant protein was analyzed and then also performed experimental challenge with live S. iniae. The result of serum agglutination titres was showed relatively low levels however, the efficacy of FKC vaccine still conferred protection against S. iniae. Moreover, the adverse effects result showed that no statistically significant difference was revealed between high concentration injected and non-injected fish groups, generally. The relative percent survival (RPS) of FKC + recombinant vaccination group was significantly higher than that of vaccinated group with FKC alone. After experimental challenge to the rock bream by injection with live bacteria (S. iniae), the FKC + rRbCC1 vaccination group was showed 87.0% RPS, however, the RPS of FKC alone vaccination was 68.2%. The results indicated that the recombinant protein as an adjuvant had a clear synergism to injection vaccine of rock bream.

Disease presentation and surgical treatment of patients with foreign-body granulomas and ASIA syndrome: case series

  • Lopez-Mendoza, Javier;Vargas-Flores, Edgar;Mouneu-Ornelas, Nicole;Altamirano-Arcos, Carlos
    • Archives of Plastic Surgery
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    • v.48 no.4
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    • pp.366-372
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    • 2021
  • Background The result of illicit polymer injection is chronic inflammation with foreign-body granuloma (FBG) formation. Treatment can be divided into medical and surgical. Some patients develop severe complications with need surgical treatment. This study aims to describe patients who underwent surgical removal of the FBGs and autoimmune/inflammatory syndrome induced by adjuvants (ASIA); additionally, we evaluated the quality of life after surgery. Methods In this retrospective single-center study, the authors examined data of patients who underwent surgical removal of FBG caused by illicit polymer injection for cosmetic purposes and confirmed ASIA from 2015 to 2020 by three different surgical approaches. Descriptive summary statistics were reported on patient demographics, presenting symptoms and clinical examination features, treatment strategies, histopathology reports and quality of life. Results The cohort included 11 female patients with FBGs and ASIA. The most affected anatomical zones were the combination of gluteal region, thighs and legs (40%); and thighs with legs (20%). Main presentation was: skin hyperpigmentation (90.9%), skin induration (63.6%), chronic fatigue (63.6%), and ulcers (36.4%). Surgical modalities consisted of: ultrasonic-assisted liposuction in four patients (36.4%); open en bloc excision and primary closure in four patients (36.4%); and open en bloc excision and microsurgical reconstruction in three patients (27.2%). The postoperative quality of life visual analog scale score was 83.9. Conclusions ASIA treatment represents a challenge for the plastic surgeon. Adequate surgical treatment emphasizing, when possible, the total or near-total resection of the FBG must be performed to improve ASIA evolution.

Comparison of immunoadjuvant activities of four bursal peptides combined with H9N2 avian influenza virus vaccine

  • Zhang, Cong;Zhou, Jiangfei;Liu, Zhixin;Liu, Yongqing;Cai, Kairui;Shen, Tengfei;Liao, Chengshui;Wang, Chen
    • Journal of Veterinary Science
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    • v.19 no.6
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    • pp.817-826
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    • 2018
  • The bursa of Fabricius (BF) is a central humoral immune organ unique to birds. Four bursal peptides (BP-I, BP-II, BP-III, and BP-IV) have been isolated and identified from the BF. In this study, the immunoadjuvant activities of BPs I to IV were examined in mice immunized with H9N2 avian influenza virus (AIV) vaccine. The results suggested that BP-I effectively enhanced cell-mediated immune responses, increased the secretion of Th1 (interferon gamma)- and Th2 (interleukin-4)-type cytokines, and induced an improved cytotoxic T-lymphocyte (CTL) response to the H9N2 virus. BP-II mainly elevated specific antibody production, especially neutralizing antibodies, and increased Th1- and Th2-type cytokine secretion. BP-III had no significant effect on antibody production or cell-mediated immune responses compared to those in the control group. A strong immune response at both the humoral and cellular levels was induced by BP-IV. Furthermore, a virus challenge experiment followed by H&E staining revealed that BP-I and BP-II promoted removal of the virus and conferred protection in mouse lungs. BP-IV significantly reduced viral titers and histopathological changes and contributed to protection against H9N2 AIV challenge in mouse lungs. This study further elucidated the immunoadjuvant activities of BPs I to IV, providing a novel insight into immunoadjuvants for use in vaccine design.

Effectiveness of Tramadol and Butorphanol as an Adjuvant to Local Anesthetic in Mandibular Nerve Block in Beagle Dogs

  • Jang, Sang Seon;Kim, Hyeonjo;Kwon, Dae Hyun;Yoon, Eunchae;Lee, Dongbin;Lee, Jae-Hoon
    • Journal of Veterinary Clinics
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    • v.39 no.5
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    • pp.226-234
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    • 2022
  • To evaluate butorphanol and tramadol as adjuvants to lidocaine in dogs undergoing mandibular nerve block. Fifteen beagles were allocated to groups based on the following treatments: lidocaine alone (L group), lidocaine + butorphanol (LB group), or lidocaine + tramadol (LT group). After mandibular nerve block with opioids as an adjunct to local anesthetics, the onset time, duration of action, and depth of anesthesia were evaluated using a quantitative method through neuromuscular blockades (NMBs) monitoring. The onset time of nerve block was 4.60 ± 2.06 min, 2.00 ± 0.00 min, and 2.60 ± 1.62 min in the L, LB, and LT groups, respectively; however, there was no statistically significant difference. The duration of nerve block was 111.88 ± 34.78 min, 302.00 ± 76.72 min, and 260.40 ± 49.88 min in the L, LB, and LT groups, respectively, with a significant difference between L and LB groups. The LB group demonstrated a more profound depth of anesthesia compared to the L and LT groups. In this study, using a quantitative method through NMBs monitoring, it was demonstrated that lidocaine and butorphanol in combination can increase the duration of nerve block and more profound the depth of anesthesia rather than lidocaine alone. Additionally, the combined use of lidocaine and opioids presented an objective indicator that could provide a more clinically stable nerve block.

A review of the immunomodulatory activities of polysaccharides isolated from Panax species

  • Hu, Yeye;He, Yang;Niu, Zhiqiang;Shen, Ting;Zhang, Ji;Wang, Xinfeng;Hu, Weicheng;Cho, Jae Youl
    • Journal of Ginseng Research
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    • v.46 no.1
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    • pp.23-32
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    • 2022
  • Panax polysaccharides are biopolymers that are isolated and purified from the roots, stems, leaves, flowers, and fruits of Panax L. plants, which have attracted considerable attention because of their immunomodulatory activities. In this paper, the composition and structural characteristics of purified polysaccharides are reviewed. Moreover, the immunomodulatory activities of polysaccharides are described both in vivo and in vitro. In vitro, Panax polysaccharides exert immunomodulatory functions mainly by activating macrophages, dendritic cells, and the complement system. In vivo, Panax polysaccharides can increase the immune organ indices and stimulate lymphocytes. In addition, this paper also discusses the membrane receptors and various signalling pathways of immune cells. Panax polysaccharides have many beneficial therapeutic effects, including enhancing or activating the immune response, and may be helpful in treating cancer, sepsis, osteoporosis, and other conditions. Panax polysaccharides have the potential for use in the development of novel therapeutic agents or adjuvants with beneficial immunomodulatory properties.