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  1 / 1519 MEDLINE  
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[PMID]:28359219
[Au] Autor:Hoppe Parr KA; Hadina S; Kilburg-Basnyat B; Wang Y; Chavez D; Thorne PS; Weiss JP
[Ad] Endereço:1 Department of Occupational and Environmental Health, The University of Iowa, Iowa City, IA, USA.
[Ti] Título:Modification of sample processing for the Limulus amebocyte lysate assay enhances detection of inflammogenic endotoxin in intact bacteria and organic dust.
[So] Source:Innate Immun;23(3):307-318, 2017 Apr.
[Is] ISSN:1753-4267
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:The pro-inflammatory potency and causal relationship with asthma of inhaled endotoxins have underscored the importance of accurately assessing the endotoxin content of organic dusts. The Limulus amebocyte lysate (LAL) assay has emerged as the preferred assay, but its ability to measure endotoxin in intact bacteria and organic dusts with similar sensitivity as purified endotoxin is unknown. We used metabolically radiolabeled Neisseria meningitidis and both rough and smooth Escherichia coli to compare dose-dependent activation in the LAL with purified endotoxin from these bacteria and shed outer membrane (OM) blebs. Labeled [ C]-3-OH-fatty acids were used to quantify the endotoxin content of the samples. Purified meningococcal and E. coli endotoxins and OM blebs displayed similar specific activity in the LAL assay to the purified LPS standard. In contrast, intact bacteria exhibited fivefold lower specific activity in the LAL assay but showed similar MD-2-dependent potency as purified endotoxin in inducing acute airway inflammation in mice. Pre-treatment of intact bacteria and organic dusts with 0.1 M Tris-HCl/10 mM EDTA increased by fivefold the release of endotoxin. These findings demonstrate that house dust and other organic dusts should be extracted with Tris/EDTA to more accurately assess the endotoxin content and pro-inflammatory potential of these environmental samples.
[Mh] Termos MeSH primário: Endotoxinas/metabolismo
Escherichia coli/imunologia
Teste do Limulus/métodos
Neisseria meningitidis/imunologia
Pneumonia/imunologia
[Mh] Termos MeSH secundário: Animais
Radioisótopos de Carbono
Erros de Diagnóstico/prevenção & controle
Poeira/análise
Endotoxinas/imunologia
Seres Humanos
Antígeno 96 de Linfócito/genética
Antígeno 96 de Linfócito/metabolismo
Camundongos
Camundongos Endogâmicos C3H
Camundongos Endogâmicos C57BL
Camundongos Knockout
Sensibilidade e Especificidade
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Carbon Radioisotopes); 0 (Dust); 0 (Endotoxins); 0 (Ly96 protein, mouse); 0 (Lymphocyte Antigen 96)
[Em] Mês de entrada:1706
[Cu] Atualização por classe:170626
[Lr] Data última revisão:
170626
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170401
[St] Status:MEDLINE
[do] DOI:10.1177/1753425917694084


  2 / 1519 MEDLINE  
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[PMID]:28185358
[Au] Autor:Taniguchi M; Ochiai A; Toyoda R; Sato T; Saitoh E; Kato T; Tanaka T
[Ad] Endereço:Department of Materials Science and Technology, Graduate School of Science and Technology, Niigata University, Niigata, 950-2181, Japan.
[Ti] Título:Effects of arginine and leucine substitutions on anti-endotoxic activities and mechanisms of action of cationic and amphipathic antimicrobial octadecapeptide from rice α-amylase.
[So] Source:J Pept Sci;23(3):252-260, 2017 Mar.
[Is] ISSN:1099-1387
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Previously, we showed that the antimicrobial cationic and amphipathic octadecapeptide AmyI-1-18 from rice α-amylase (AmyI-1) inhibited the endotoxic activity of lipopolysaccharide (LPS) from Escherichia coli. In addition, we demonstrated that several AmyI-1-18 analogs containing arginine or leucine substitutions, which were designed on the basis of the helical wheel projection of AmyI-1-18, exhibited higher antimicrobial activity against human pathogenic microorganisms than AmyI-1-18. In the present study, anti-inflammatory (anti-endotoxic) activities of five AmyI-1-18 analogs containing arginine or leucine substitutions were investigated. Two single arginine-substituted and two single leucine-substituted AmyI-1-18 analogs inhibited the production of LPS-induced nitric oxide in mouse macrophages (RAW264) more effectively than AmyI-1-18. These data indicate that enhanced cationic and hydrophobic properties of AmyI-1-18 are associated with improved anti-endotoxic activity. In subsequent chromogenic Limulus amebocyte lysate assays, 50% inhibitory concentrations (IC ) of the three AmyI-1-18 analogs (G12R, D15R, and E9L) were 0.11-0.13 µm, indicating higher anti-endotoxic activity than that of AmyI-1-18 (IC 0.22 µm), and specific LPS binding activity. In agreement, surface plasmon resonance analyses confirmed direct LPS binding of three AmyI-1-18 analogs. In addition, AmyI-1-18 analogs exhibited little or no cytotoxic activity against RAW264 cells, indicating that enhancements of anti-inflammatory and LPS-neutralizing activities following replacement of arginine or leucine did not result in significant increases in cytotoxicity. This study shows that the arginine-substituted and leucine-substituted AmyI-1-18 analogs with improved anti-endotoxic and antimicrobial activities have clinical potential as dual-function host defense agents. Copyright © 2017 European Peptide Society and John Wiley & Sons, Ltd.
[Mh] Termos MeSH primário: Anti-Inflamatórios não Esteroides/farmacologia
Peptídeos Catiônicos Antimicrobianos/farmacologia
Arginina/química
Leucina/química
Lipopolissacarídeos/antagonistas & inibidores
Proteínas de Plantas/farmacologia
alfa-Amilases/farmacologia
[Mh] Termos MeSH secundário: Sequência de Aminoácidos
Substituição de Aminoácidos
Animais
Anti-Inflamatórios não Esteroides/síntese química
Anti-Inflamatórios não Esteroides/química
Peptídeos Catiônicos Antimicrobianos/síntese química
Peptídeos Catiônicos Antimicrobianos/química
Linhagem Celular
Seres Humanos
Interações Hidrofóbicas e Hidrofílicas
Teste do Limulus
Lipopolissacarídeos/química
Lipopolissacarídeos/farmacologia
Macrófagos/citologia
Macrófagos/efeitos dos fármacos
Macrófagos/metabolismo
Camundongos
Óxido Nítrico/antagonistas & inibidores
Óxido Nítrico/biossíntese
Oryza/química
Proteínas de Plantas/síntese química
Proteínas de Plantas/química
Ligação Proteica
Relação Estrutura-Atividade
alfa-Amilases/síntese química
alfa-Amilases/química
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Anti-Inflammatory Agents, Non-Steroidal); 0 (Antimicrobial Cationic Peptides); 0 (Lipopolysaccharides); 0 (Plant Proteins); 31C4KY9ESH (Nitric Oxide); 94ZLA3W45F (Arginine); EC 3.2.1.1 (alpha-Amylases); GMW67QNF9C (Leucine)
[Em] Mês de entrada:1703
[Cu] Atualização por classe:170307
[Lr] Data última revisão:
170307
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170211
[St] Status:MEDLINE
[do] DOI:10.1002/psc.2983


  3 / 1519 MEDLINE  
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[PMID]:27913792
[Au] Autor:Mizumura H; Ogura N; Aketagawa J; Aizawa M; Kobayashi Y; Kawabata SI; Oda T
[Ad] Endereço:1 LAL Research and Development Department, Seikagaku Corporation, Tokyo, Japan.
[Ti] Título:Genetic engineering approach to develop next-generation reagents for endotoxin quantification.
[So] Source:Innate Immun;23(2):136-146, 2017 Feb.
[Is] ISSN:1753-4267
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:The bacterial endotoxin test, which uses amebocyte lysate reagents of horseshoe crab origin, is a sensitive, reproducible and simple assay to measure endotoxin concentration. To develop sustainable raw materials for lysate reagents that do not require horseshoe crabs, three recombinant protease zymogens (factor C, derived from mammalian cells; factor B; and the proclotting enzyme derived from insect cells) were prepared using a genetic engineering technique. Recombinant cascade reagents (RCRs) were then prepared to reconstruct the reaction cascade in the amebocyte lysate reagent. The protease activity of the RCR containing recombinant factor C was much greater than that of recombinant factor C alone, indicating the efficiency of signal amplification in the cascade. Compared with the RCR containing the insect cell-derived factor C, those containing mammalian cell-derived factor C, which features different glycosylation patterns, were less susceptible to interference by the injectable drug components. The standard curve of the RCR containing mammalian cell-derived recombinant factor C had a steeper slope than the curves for those containing natural lysate reagents, suggesting a greater sensitivity to endotoxin. The present study supports the future production of recombinant reagents that do not require the use of natural resources.
[Mh] Termos MeSH primário: Fator B do Complemento/metabolismo
Endopeptidases/metabolismo
Endotoxinas/análise
Precursores Enzimáticos/metabolismo
Proteínas de Insetos/metabolismo
Teste do Limulus/métodos
Serina Endopeptidases/metabolismo
[Mh] Termos MeSH secundário: Animais
Extratos Celulares
Fator B do Complemento/genética
Endopeptidases/genética
Precursores Enzimáticos/genética
Engenharia Genética
Caranguejos Ferradura
Indicadores e Reagentes
Proteínas de Insetos/genética
Proteínas Recombinantes/genética
Padrões de Referência
Sensibilidade e Especificidade
Serina Endopeptidases/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Cell Extracts); 0 (Endotoxins); 0 (Enzyme Precursors); 0 (Indicators and Reagents); 0 (Insect Proteins); 0 (Recombinant Proteins); EC 3.4.- (Endopeptidases); EC 3.4.21.- (Serine Endopeptidases); EC 3.4.21.47 (Complement Factor B); EC 3.4.99.- (pro-clotting enzyme)
[Em] Mês de entrada:1706
[Cu] Atualização por classe:170629
[Lr] Data última revisão:
170629
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161204
[St] Status:MEDLINE
[do] DOI:10.1177/1753425916681074


  4 / 1519 MEDLINE  
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[PMID]:27880838
[Au] Autor:Mukherjee SP; Lozano N; Kucki M; Del Rio-Castillo AE; Newman L; Vázquez E; Kostarelos K; Wick P; Fadeel B
[Ad] Endereço:Nanosafety & Nanomedicine Laboratory, Division of Molecular Toxicology, Institute of Environmental Medicine, Karolinska Institutet, Stockholm, Sweden.
[Ti] Título:Detection of Endotoxin Contamination of Graphene Based Materials Using the TNF-α Expression Test and Guidelines for Endotoxin-Free Graphene Oxide Production.
[So] Source:PLoS One;11(11):e0166816, 2016.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Nanomaterials may be contaminated with bacterial endotoxin during production and handling, which may confound toxicological testing of these materials, not least when assessing for immunotoxicity. In the present study, we evaluated the conventional Limulus amebocyte lysate (LAL) assay for endotoxin detection in graphene based material (GBM) samples, including graphene oxide (GO) and few-layered graphene (FLG). Our results showed that some GO samples interfered with various formats of the LAL assay. To overcome this problem, we developed a TNF-α expression test (TET) using primary human monocyte-derived macrophages incubated in the presence or absence of the endotoxin inhibitor, polymyxin B sulfate, and found that this assay, performed with non-cytotoxic doses of the GBM samples, enabled unequivocal detection of endotoxin with a sensitivity that is comparable to the LAL assay. FLG also triggered TNF-α production in the presence of the LPS inhibitor, pointing to an intrinsic pro-inflammatory effect. Finally, we present guidelines for the preparation of endotoxin-free GO, validated by using the TET.
[Mh] Termos MeSH primário: Bioensaio/métodos
Endotoxinas/análise
Grafite/química
Fator de Necrose Tumoral alfa/análise
[Mh] Termos MeSH secundário: Células Cultivadas
Endotoxinas/antagonistas & inibidores
Endotoxinas/metabolismo
Ensaio de Imunoadsorção Enzimática
Guias como Assunto
Seres Humanos
Teste do Limulus
Macrófagos/citologia
Macrófagos/efeitos dos fármacos
Macrófagos/metabolismo
Óxidos/química
Polimixina B/química
Polimixina B/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Endotoxins); 0 (Oxides); 0 (Tumor Necrosis Factor-alpha); 1404-26-8 (Polymyxin B); 7782-42-5 (Graphite)
[Em] Mês de entrada:1706
[Cu] Atualização por classe:170627
[Lr] Data última revisão:
170627
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161124
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0166816


  5 / 1519 MEDLINE  
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[PMID]:27854308
[Au] Autor:Park HJ; Oh S; Vinod N; Ji S; Noh HB; Koo JM; Lee SH; Kim SC; Lee KS; Choi CW
[Ad] Endereço:Department of Biology & Medicinal Science, Pai Chai University, Daejeon 35345, Korea. parkhj0524@pcu.ac.kr.
[Ti] Título:Characterization of Chemically-Induced Bacterial Ghosts (BGs) Using Sodium Hydroxide-Induced Vibrio parahaemolyticus Ghosts (VPGs).
[So] Source:Int J Mol Sci;17(11), 2016 Nov 15.
[Is] ISSN:1422-0067
[Cp] País de publicação:Switzerland
[La] Idioma:eng
[Ab] Resumo:Acellular bacterial ghosts (BGs) are empty non-living bacterial cell envelopes, commonly generated by controlled expression of the cloned lysis gene of bacteriophage PhiX174. In this study, ghosts (VPGs) were generated by chemically-induced lysis and the method is based on minimum inhibitory concentration (MIC) of sodium hydroxide (NaOH), acetic acid, boric acid, citric acid, maleic acid, hydrochloric acid, and sulfuric acid. The MIC values of the respective chemicals were 3.125, 6.25, <50.0, 25.0, 6.25, 1.56, and 0.781 mg/mL. Except for boric acid, the lysis efficiency reached more than 99.99% at 5 min after treatment of all chemicals. Among those chemicals, NaOH-induced VPGs appeared completely DNA-free, which was confirmed by quantitative real-time PCR. Besides, lipopolysaccharides (LPS) extracted from the NaOH-induced VPGs showed no distinctive band on SDS-PAGE gel after silver staining. On the other hand, LPS extracted from wild-type bacterial cells, as well as the organic acids-induced VPGs showed triple major bands and LPS extracted from the inorganic acids-induced VPGs showed double bands. It suggests that some surface structures in LPS of the NaOH-induced VPGs may be lost, weakened, or modified by the MIC of NaOH. Nevertheless, lysate assay revealed that there is no significant difference in endotoxic activity between the NaOH-induced VPGs and wild-type bacterial cells. Macrophages exposed to the NaOH-induced VPGs at 0.5 × 106 CFU/mL showed cell viability of 97.9%, however, the MIC of NaOH did not reduce the cytotoxic effect of wild-type bacterial cells. Like LPS, the NaOH-induced VPGs are an excellent activator of pro-inflammatory cytokines (IL-1ß and iNOS), anti-inflammatory cytokine (IL-10), and dual activities (IL-6) in the stimulated macrophage cells. On the other hand, the induction of TNF-α mRNA was remarkable in the macrophages exposed with wild-type cells. Scanning electron microscopy showed the formation of trans-membrane lysis tunnel structures in the NaOH-induced VPGs. SDS-PAGE and agarose gel electrophoresis also confirmed that cytoplasmic proteins and genomic DNA released from the VPGs to culture medium through the lysis tunnel structures. Taken together, all these data indicate that the NaOH-induced VPGs show the potency of a safe, economical, and effective inactivated bacterial vaccine candidate.
[Mh] Termos MeSH primário: Membrana Celular/química
DNA Bacteriano/secreção
Macrófagos/efeitos dos fármacos
Hidróxido de Sódio/farmacologia
[Mh] Termos MeSH secundário: Ácido Acético/farmacologia
Animais
Ácidos Bóricos/farmacologia
Linhagem Celular
Membrana Celular/imunologia
Sobrevivência Celular/efeitos dos fármacos
Ácido Cítrico/farmacologia
Expressão Gênica
Ácido Clorídrico/farmacologia
Interleucina-10/genética
Interleucina-10/imunologia
Interleucina-1beta/genética
Interleucina-1beta/imunologia
Interleucina-6/genética
Interleucina-6/imunologia
Teste do Limulus
Lipopolissacarídeos/química
Lipopolissacarídeos/metabolismo
Macrófagos/citologia
Macrófagos/imunologia
Maleatos/farmacologia
Camundongos
Testes de Sensibilidade Microbiana
Óxido Nítrico Sintase Tipo II/genética
Óxido Nítrico Sintase Tipo II/imunologia
Ácidos Sulfúricos/farmacologia
Fator de Necrose Tumoral alfa/genética
Fator de Necrose Tumoral alfa/imunologia
Vibrio parahaemolyticus/química
Vibrio parahaemolyticus/efeitos dos fármacos
Vibrio parahaemolyticus/imunologia
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Boric Acids); 0 (DNA, Bacterial); 0 (IL10 protein, mouse); 0 (IL1B protein, mouse); 0 (Interleukin-1beta); 0 (Interleukin-6); 0 (Lipopolysaccharides); 0 (Maleates); 0 (Sulfuric Acids); 0 (Tumor Necrosis Factor-alpha); 0 (interleukin-6, mouse); 130068-27-8 (Interleukin-10); 2968PHW8QP (Citric Acid); 55X04QC32I (Sodium Hydroxide); 91XW058U2C (maleic acid); EC 1.14.13.39 (Nitric Oxide Synthase Type II); EC 1.14.13.39 (Nos2 protein, mouse); O40UQP6WCF (sulfuric acid); Q40Q9N063P (Acetic Acid); QTT17582CB (Hydrochloric Acid); R57ZHV85D4 (boric acid)
[Em] Mês de entrada:1703
[Cu] Atualização por classe:170328
[Lr] Data última revisão:
170328
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161118
[St] Status:MEDLINE


  6 / 1519 MEDLINE  
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[PMID]:27764208
[Au] Autor:Wong J; Zhang Y; Patidar A; Vilar E; Finkelman M; Farrington K
[Ad] Endereço:Lister Renal Unit, Hertfordshire, United Kingdom.
[Ti] Título:Is Endotoxemia in Stable Hemodialysis Patients an Artefact? Limitations of the Limulus Amebocyte Lysate Assay and Role of (1→3)-ß-D Glucan.
[So] Source:PLoS One;11(10):e0164978, 2016.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:BACKGROUND: Elevated blood endotoxin levels are frequently reported in the dialysis population and are strongly linked with inflammation, a major predictor of mortality. Virtually all studies have employed the Limulus Amoebocyte Lysate (LAL) assay to detect endotoxin. However this assay is not endotoxin-specific and can be activated by (1→3)-ß-glucan (BG), a component of fungal cell walls leading to false positive signals. Very few studies have taken account of this. We examined the influence of BG-based activation of the LAL assay on the detection of endotoxemia in this setting. METHOD: We measured plasma endotoxin levels in 50 hemodialysis patients with and without the use of BG-blocking buffers. These buffers inhibit BG activation of the LAL assay to ensure that any signal detected is endotoxin-specific. Blood samples were measured for BG, interleukin-6 (IL-6), tumor necrosis factor-alfa (TNF-α) to examine the association between endotoxin signals, BG and inflammation. RESULTS: Endotoxin signals were detected in 50% of patients. On repeat measurement with a BG-blocking buffer, all detected endotoxin signals were extinguished. No patient had detectable endotoxemia. Plasma BG levels were significantly elevated in 58% of patients and were higher in those with detectable endotoxin signals using the LAL assay without BG-blocking buffers (78vs.54pg/mL;p<0.001). Endotoxin signal and BG levels did not correlate with levels of TNF-α or IL-6. CONCLUSION: Use of the LAL assay for blood endotoxin detection in dialysis patients has its limitations due to high blood BG. Endotoxemia frequently reported in non-infected hemodialysis patients may be artefactual due to BG interference.
[Mh] Termos MeSH primário: Artefatos
Endotoxemia/sangue
Teste do Limulus/métodos
Diálise Renal
beta-Glucanas/sangue
[Mh] Termos MeSH secundário: Idoso
Tampões (Química)
Estudos Transversais
Endotoxemia/diagnóstico
Endotoxemia/etiologia
Endotoxinas/sangue
Reações Falso-Positivas
Feminino
Seres Humanos
Masculino
Meia-Idade
Diálise Renal/efeitos adversos
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Buffers); 0 (Endotoxins); 0 (beta-Glucans)
[Em] Mês de entrada:1706
[Cu] Atualização por classe:170606
[Lr] Data última revisão:
170606
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161021
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0164978


  7 / 1519 MEDLINE  
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[PMID]:27494624
[Au] Autor:Fennrich S; Hennig U; Toliashvili L; Schlensak C; Wendel HP; Stoppelkamp S
[Ad] Endereço:Clinical Research Laboratory, Clinic of Thoracic, Cardiac and Vascular Surgery, University Hospital Tübingen, Germany.
[Ti] Título:More than 70 years of pyrogen detection: Current state and future perspectives.
[So] Source:Altern Lab Anim;44(3):239-53, 2016 Jul.
[Is] ISSN:0261-1929
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:In the quality assurance of medical products, tests for sterility are essential. For parenteral pharmaceuticals, avoiding the presence of pyrogens is crucial. These fever-inducing substances (endotoxins and non-endotoxins) are not eliminated by standard sterilisation processes, and are biologically active once in the bloodstream, causing risks to human health, ranging from mild reactions (e.g. fever) to septic shock and death. Therefore, for injectable formulations, pyrogen testing is mandatory. Over the years, various pyrogen testing methods have been introduced, namely: in the 1940s, the rabbit pyrogen test, which is an in vivo test that measures the fever reaction as an endpoint; in the 1970s, the Limulus Amoebocyte Lysate (LAL) test, which is an in vitro test (with the haemolymph of the horseshoe crab) that specifically detects endotoxin; and in 2010, the Monocyte-Activation Test (MAT), which is a non-animal based in vitro pyrogen test that represents a full replacement of the rabbit test. Due to the ubiquity and biological significance of pyrogens, we are currently further developing the MAT so that it can be used for other applications. More specifically, our focus is on the detection of pyrogenic contamination on medical devices, as well as on the measurement of air quality. In addition, further improvements to permit the use of cryopreserved blood in the MAT, to overcome the limitations in the availability of freshly-drawn blood from human donors, are ongoing.
[Mh] Termos MeSH primário: Alternativas aos Testes com Animais/métodos
Teste do Limulus/história
Pirogênios/isolamento & purificação
[Mh] Termos MeSH secundário: Alternativas aos Testes com Animais/tendências
Animais
Citocinas/genética
Citocinas/metabolismo
Regulação da Expressão Gênica/efeitos dos fármacos
História do Século XX
História do Século XXI
Caranguejos Ferradura/metabolismo
Seres Humanos
Monócitos/efeitos dos fármacos
Monócitos/metabolismo
Pirogênios/toxicidade
Coelhos
[Pt] Tipo de publicação:HISTORICAL ARTICLE; JOURNAL ARTICLE; REVIEW
[Nm] Nome de substância:
0 (Cytokines); 0 (Pyrogens)
[Em] Mês de entrada:1701
[Cu] Atualização por classe:170107
[Lr] Data última revisão:
170107
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:160806
[St] Status:MEDLINE


  8 / 1519 MEDLINE  
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[PMID]:27470947
[Au] Autor:Bolden JS; Warburton RE; Phelan R; Murphy M; Smith KR; De Felippis MR; Chen D
[Ad] Endereço:Global Quality Laboratories, Eli Lilly and Company, Indianapolis, IN, 46285, USA.
[Ti] Título:Endotoxin recovery using limulus amebocyte lysate (LAL) assay.
[So] Source:Biologicals;44(5):434-40, 2016 Sep.
[Is] ISSN:1095-8320
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:A phenomenon initially reported by Chen and Vinther in 2013 [1], and now commonly referred to as low endotoxin recovery (LER), has prompted the Food and Drug Administration (FDA) to request specific data demonstrating the capability of the LAL BET method (i.e., USP <85>) to recover endotoxin from spiked samples over time. The results of these spike/hold recovery studies are expected to be included in the Biologics License Applications (BLA) for review by the Center for Drug Evaluation and Research (CDER) Hughes (2014) and Hughes et al. (2015) [2,3]. Such studies involve spiking a known amount of a surrogate endotoxin, such as purified lipopolysaccharide (LPS), into undiluted biological products and then testing at different time points to determine the recovery over time. We report here the experience and learning gained from conducting spike/hold recovery studies for a monoclonal antibody (Mab) product. Results from initial hold studies spiked with purified LPS showed rapid loss of endotoxin activity in the drug substance (DS) and significant batch-to-batch variation in the drug product (DP). After careful review and examination of the experimental details, it was determined that the study design and execution differed from the routine batch release USP <85> BET method with regard to mixing time and sampling scheme. The hold study design was subsequently revised so that the mixing time and sampling were the same as the verified USP <85> BET method used for routine batch release testing. The spike/hold recovery studies were repeated and the results demonstrated that LPS could be consistently recovered over time. These findings highlight the importance of carefully controlling sample preparation procedures in a spike/hold recovery study in order to demonstrate the suitability of using the LAL BET method for endotoxin detection.
[Mh] Termos MeSH primário: Teste do Limulus/métodos
Lipopolissacarídeos/análise
[Mh] Termos MeSH secundário: Lipopolissacarídeos/química
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Lipopolysaccharides)
[Em] Mês de entrada:1701
[Cu] Atualização por classe:170123
[Lr] Data última revisão:
170123
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:160730
[St] Status:MEDLINE


  9 / 1519 MEDLINE  
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[PMID]:27464990
[Au] Autor:Reich J; Lang P; Grallert H; Motschmann H
[Ad] Endereço:Institute of Physical and Theoretical Chemistry, University of Regensburg, Regensburg, Germany. Electronic address: reich.johannes@icloud.com.
[Ti] Título:Masking of endotoxin in surfactant samples: Effects on Limulus-based detection systems.
[So] Source:Biologicals;44(5):417-22, 2016 Sep.
[Is] ISSN:1095-8320
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Over the last few decades Limulus Amebocyte Lysate (LAL) has been the most sensitive method for the detection of endotoxins (Lipopolysaccharides) and is well accepted in a broad field of applications. Recently, Low Endotoxin Recovery (LER) in biopharmaceutical drug products has been noticed, whereby the detection of potential endotoxin contaminations is not ensured. Notably, most of these drug products contain surfactants, which can have crucial effects on the detectability of endotoxin. In order to analyze the driving forces of LER, endotoxin detection in samples containing nonionic surfactants in various buffer systems was investigated. The results show that the process of LER is kinetically controlled and temperature-dependent. Furthermore, only the simultaneous presence of nonionic surfactants and components capable of forming metal complexes resulted in LER. In addition, capacity experiments show that even hazardous amounts of endotoxin can remain undetectable within such formulation compositions. In conclusion, the LER phenomenon is caused by endotoxin masking and not by test interference. In this process, the supramolecular structure of endotoxin is altered and exhibits only a limited susceptibility in binding to the Factor C of Limulus-based detection systems. We propose a two-step mechanism of endotoxin masking by complex forming agents and nonionic surfactants.
[Mh] Termos MeSH primário: Teste do Limulus/métodos
Lipopolissacarídeos/análise
Tensoativos/química
[Mh] Termos MeSH secundário: Lipopolissacarídeos/química
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Lipopolysaccharides); 0 (Surface-Active Agents)
[Em] Mês de entrada:1701
[Cu] Atualização por classe:170123
[Lr] Data última revisão:
170123
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:160729
[St] Status:MEDLINE


  10 / 1519 MEDLINE  
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[PMID]:27418356
[Au] Autor:Munford RS
[Ad] Endereço:Laboratory of Clinical Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 9000 Rockville Pike, Bethesda, MD, USA munfordrs@niaid.nih.gov.
[Ti] Título:Endotoxemia-menace, marker, or mistake?
[So] Source:J Leukoc Biol;100(4):687-698, 2016 Oct.
[Is] ISSN:1938-3673
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Endotoxemia is in its scientific ascendancy. Never has blood-borne, Gram-negative bacterial endotoxin (LPS) been invoked in the pathogenesis of so many diseases-not only as a trigger for septic shock, once its most cited role, but also as a contributor to atherosclerosis, obesity, chronic fatigue, metabolic syndrome, and many other conditions. Finding elevated plasma endotoxin levels has been essential supporting evidence for each of these links, yet the assays used to detect and quantitate endotoxin have important limitations. This article describes several assays for endotoxin in plasma, reviews what they do and do not measure, and discusses why LPS heterogeneity, LPS trafficking pathways, and host LPS inactivation mechanisms should be considered when interpreting endotoxin assay results.
[Mh] Termos MeSH primário: Endotoxemia
[Mh] Termos MeSH secundário: Transporte Biológico
Hidrolases de Éster Carboxílico/metabolismo
Endotoxemia/sangue
Endotoxemia/complicações
Ensaio de Imunoadsorção Enzimática
Bactérias Gram-Negativas/química
Bactérias Gram-Negativas/patogenicidade
Infecções por Bactérias Gram-Negativas/sangue
Infecções por Bactérias Gram-Negativas/complicações
Seres Humanos
Absorção Intestinal
Teste do Limulus
Lipídeo A/sangue
Lipídeo A/química
Lipopolissacarídeos/sangue
Lipopolissacarídeos/química
Linfa/metabolismo
Testes de Neutralização
Antígenos O/sangue
Antígenos O/química
Relação Estrutura-Atividade
[Pt] Tipo de publicação:JOURNAL ARTICLE; REVIEW
[Nm] Nome de substância:
0 (Lipid A); 0 (Lipopolysaccharides); 0 (O Antigens); EC 3.1.1.- (Carboxylic Ester Hydrolases); EC 3.1.1.- (acyloxyacyl hydrolase)
[Em] Mês de entrada:1709
[Cu] Atualização por classe:171001
[Lr] Data última revisão:
171001
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:160716
[St] Status:MEDLINE



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