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[PMID]:29020090
[Au] Autor:Leitman EM; Palmer CD; Buus S; Chen F; Riddell L; Sims S; Klenerman P; Sáez-Cirión A; Walker BD; Hess PR; Altfeld M; Matthews PC; Goulder PJR
[Ad] Endereço:Department of Paediatrics, University of Oxford, Oxford, United Kingdom.
[Ti] Título:Saporin-conjugated tetramers identify efficacious anti-HIV CD8+ T-cell specificities.
[So] Source:PLoS One;12(10):e0184496, 2017.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Antigen-specific T-cells are highly variable, spanning potent antiviral efficacy and damaging auto-reactivity. In virus infections, identifying the most efficacious responses is critical to vaccine design. However, current methods depend on indirect measures or on ex vivo expanded CTL clones. We here describe a novel application of cytotoxic saporin-conjugated tetramers to kill antigen-specific T-cells without significant off-target effects. The relative efficacy of distinct antiviral CD8+ T-cell specificity can be directly assessed via antigen-specific CD8+ T-cell depletion. The utility of these reagents is demonstrated here in identifying the CD8+ T-cell specificity most effective in preventing HIV progression in HIV-infected HLA-B*27-positive immune controllers.
[Mh] Termos MeSH primário: Linfócitos T CD8-Positivos/imunologia
Infecções por HIV/tratamento farmacológico
Infecções por HIV/imunologia
Multimerização Proteica
Proteínas Inativadoras de Ribossomos Tipo 1/uso terapêutico
[Mh] Termos MeSH secundário: Antivirais/farmacologia
Linfócitos T CD8-Positivos/efeitos dos fármacos
Endocitose/efeitos dos fármacos
Seres Humanos
Depleção Linfocítica
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Antiviral Agents); 0 (Ribosome Inactivating Proteins, Type 1); EC 3.2.2.22 (saporin)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171101
[Lr] Data última revisão:
171101
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171012
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0184496


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[PMID]:28705805
[Au] Autor:Diepenbroek C; Quinn D; Stephens R; Zollinger B; Anderson S; Pan A; de Lartigue G
[Ad] Endereço:The John B. Pierce Laboratory, New Haven, Connecticut.
[Ti] Título:Validation and characterization of a novel method for selective vagal deafferentation of the gut.
[So] Source:Am J Physiol Gastrointest Liver Physiol;313(4):G342-G352, 2017 Oct 01.
[Is] ISSN:1522-1547
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:There is a lack of tools that selectively target vagal afferent neurons (VAN) innervating the gut. We use saporin (SAP), a potent neurotoxin, conjugated to the gastronintestinal (GI) hormone cholecystokinin (CCK-SAP) injected into the nodose ganglia (NG) of male Wistar rats to specifically ablate GI-VAN. We report that CCK-SAP ablates a subpopulation of VAN in culture. In vivo, CCK-SAP injection into the NG reduces VAN innervating the mucosal and muscular layers of the stomach and small intestine but not the colon, while leaving vagal efferent neurons intact. CCK-SAP abolishes feeding-induced c-Fos in the NTS, as well as satiation by CCK or glucagon like peptide-1 (GLP-1). CCK-SAP in the NG of mice also abolishes CCK-induced satiation. Therefore, we provide multiple lines of evidence that injection of CCK-SAP in NG is a novel selective vagal deafferentation technique of the upper GI tract that works in multiple vertebrate models. This method provides improved tissue specificity and superior separation of afferent and efferent signaling compared with vagotomy, capsaicin, and subdiaphragmatic deafferentation. We develop a new method that allows targeted lesioning of vagal afferent neurons that innervate the upper GI tract while sparing vagal efferent neurons. This reliable approach provides superior tissue specificity and selectivity for vagal afferent over efferent targeting than traditional approaches. It can be used to address questions about the role of gut to brain signaling in physiological and pathophysiological conditions.
[Mh] Termos MeSH primário: Vias Aferentes/efeitos dos fármacos
Denervação Autônoma/métodos
Trato Gastrointestinal/efeitos dos fármacos
Bloqueio Nervoso/métodos
Proteínas Inativadoras de Ribossomos Tipo 1/administração & dosagem
Nervo Vago/efeitos dos fármacos
[Mh] Termos MeSH secundário: Vias Aferentes/fisiologia
Animais
Trato Gastrointestinal/fisiologia
Masculino
Neurotoxinas/administração & dosagem
Neurotoxinas/farmacologia
Ratos
Ratos Wistar
Resultado do Tratamento
Nervo Vago/fisiologia
[Pt] Tipo de publicação:JOURNAL ARTICLE; VALIDATION STUDIES
[Nm] Nome de substância:
0 (Neurotoxins); 0 (Ribosome Inactivating Proteins, Type 1); EC 3.2.2.22 (saporin)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171108
[Lr] Data última revisão:
171108
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170715
[St] Status:MEDLINE
[do] DOI:10.1152/ajpgi.00095.2017


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[PMID]:28320279
[Au] Autor:Bondarenko NS; Dilmukhametova LK; Kurina AY; Murtazina AR; Sapronova AY; Sysoeva AP; Ugrumov MV
[Ad] Endereço:Koltzov Institute of Developmental Biology, Russian Academy of Sciences, Moscow, 119334, Russia. aliya_mr89@mail.ru.
[Ti] Título:Plasticity of Central and Peripheral Sources of Noradrenaline in Rats during Ontogenesis.
[So] Source:Biochemistry (Mosc);82(3):373-379, 2017 Mar.
[Is] ISSN:1608-3040
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:The morphogenesis of individual organs and the whole organism occurs under the control of intercellular chemical signals mainly during the perinatal period of ontogenesis in rodents. In this study, we tested our hypothesis that the biologically active concentration of noradrenaline (NA) in blood in perinatal ontogenesis of rats is maintained due to humoral interaction between its central and peripheral sources based on their plasticity. As one of the mechanisms of plasticity, we examined changes in the secretory activity (spontaneous and stimulated release of NA) of NA-producing organs under deficiency of its synthesis in the brain. The destruction of NA-ergic neurons was provoked by administration of a hybrid molecular complex - antibodies against dopamine-ß-hydroxylase associated with the cytotoxin saporin - into the lateral cerebral ventricles of neonatal rats. We found that 72 h after the inhibition of NA synthesis in the brain, its spontaneous release from hypothalamus increased, which was most likely due to a compensatory increase of NA secretion from surviving neurons and can be considered as one of the mechanisms of neuroplasticity aimed at the maintenance of its physiological concentration in peripheral blood. Noradrenaline secretion from peripheral sources (adrenal glands and the organ of Zuckerkandl) also showed a compensatory increase in this model. Thus, during the critical period of morphogenesis, the brain is integrated into the system of NA-producing organs and participates in their reciprocal humoral regulation as manifested in compensatory enhancement of NA secretion in each of the studied sources of NA under specific inhibition of NA production in the brain.
[Mh] Termos MeSH primário: Glândulas Suprarrenais
Neurônios Adrenérgicos/secreção
Ventrículos Cerebrais
Hipotálamo
Norepinefrina/secreção
Glomos Para-Aórticos/secreção
[Mh] Termos MeSH secundário: Glândulas Suprarrenais/crescimento & desenvolvimento
Glândulas Suprarrenais/secreção
Animais
Ventrículos Cerebrais/crescimento & desenvolvimento
Ventrículos Cerebrais/secreção
Dopamina beta-Hidroxilase/metabolismo
Hipotálamo/crescimento & desenvolvimento
Hipotálamo/secreção
Masculino
Ratos
Ratos Wistar
Proteínas Inativadoras de Ribossomos Tipo 1/toxicidade
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Ribosome Inactivating Proteins, Type 1); EC 1.14.17.1 (Dopamine beta-Hydroxylase); EC 3.2.2.22 (saporin); X4W3ENH1CV (Norepinephrine)
[Em] Mês de entrada:1703
[Cu] Atualização por classe:170322
[Lr] Data última revisão:
170322
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170322
[St] Status:MEDLINE
[do] DOI:10.1134/S0006297917030166


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[PMID]:28264667
[Au] Autor:Jeong DU; Lee J; Chang WS; Chang JW
[Ad] Endereço:Brain Korea 21 PLUS Project for Medical Science and Brain Research Institute, Yonsei University College of Medicine, Seoul, Korea.
[Ti] Título:Identifying the appropriate time for deep brain stimulation to achieve spatial memory improvement on the Morris water maze.
[So] Source:BMC Neurosci;18(1):29, 2017 Mar 07.
[Is] ISSN:1471-2202
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:BACKGROUND: The possibility of using deep brain stimulation (DBS) for memory enhancement has recently been reported, but the precise underlying mechanisms of its effects remain unknown. Our previous study suggested that spatial memory improvement by medial septum (MS)-DBS may be associated with cholinergic regulation and neurogenesis. However, the affected stage of memory could not be distinguished because the stimulation was delivered during the execution of all memory processes. Therefore, this study was performed to determine the stage of memory affected by MS-DBS. Rats were administered 192 IgG-saporin to lesion cholinergic neurons. Stimulation was delivered at different times in different groups of rats: 5 days before the Morris water maze test (pre-stimulation), 5 days during the training phase of the Morris water maze test (training-stimulation), and 2 h before the Morris water maze probe test (probe-stimulation). A fourth group of rats was lesioned but received no stimulation. These four groups were compared with a normal (control) group. RESULTS: The most effective memory restoration occurred in the pre-stimulation group. Moreover, the pre-stimulation group exhibited better recall of the platform position than the other stimulation groups. An increase in the level of brain derived neurotrophic factor (BDNF) was observed in the pre-stimulation group; this increase was maintained for 1 week. However, acetylcholinesterase activity in the pre-stimulation group was not significantly different from the lesion group. CONCLUSION: Memory impairment due to cholinergic denervation can be improved by DBS. The improvement is significantly correlated with the up-regulation of BDNF expression and neurogenesis. Based on the results of this study, the use of MS-DBS during the early stage of disease may restore spatial memory impairment.
[Mh] Termos MeSH primário: Estimulação Encefálica Profunda/métodos
Aprendizagem em Labirinto/fisiologia
Núcleos Septais/fisiologia
Memória Espacial/fisiologia
[Mh] Termos MeSH secundário: Animais
Anticorpos Monoclonais/administração & dosagem
Fator Neurotrófico Derivado do Encéfalo/metabolismo
Colina O-Acetiltransferase/metabolismo
Colinérgicos/administração & dosagem
Neurônios Colinérgicos/efeitos dos fármacos
Neurônios Colinérgicos/metabolismo
Glutamato Descarboxilase/metabolismo
Masculino
Neurogênese
Ratos
Ratos Sprague-Dawley
Proteínas Inativadoras de Ribossomos Tipo 1/administração & dosagem
Núcleos Septais/efeitos dos fármacos
Núcleos Septais/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (192 IgG-saporin); 0 (Antibodies, Monoclonal); 0 (Brain-Derived Neurotrophic Factor); 0 (Cholinergic Agents); 0 (Ribosome Inactivating Proteins, Type 1); EC 2.3.1.6 (Choline O-Acetyltransferase); EC 4.1.1.15 (Glutamate Decarboxylase); EC 4.1.1.15 (glutamate decarboxylase 1); EC 4.1.1.15 (glutamate decarboxylase 2)
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170915
[Lr] Data última revisão:
170915
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170308
[St] Status:MEDLINE
[do] DOI:10.1186/s12868-017-0345-4


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[PMID]:28235471
[Au] Autor:Smith WS; Baker EJ; Holmes SE; Koster G; Hunt AN; Johnston DA; Flavell SU; Flavell DJ
[Ad] Endereço:The Simon Flavell Leukaemia Research Laboratory, Southampton General Hospital, Southampton, Hampshire SO16 6YD, United Kingdom.
[Ti] Título:Membrane cholesterol is essential for triterpenoid saponin augmentation of a saporin-based immunotoxin directed against CD19 on human lymphoma cells.
[So] Source:Biochim Biophys Acta;1859(5):993-1007, 2017 05.
[Is] ISSN:0006-3002
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:Triterpenoid saponins from Saponinum Album (SA) exert potent lytic effects on eukaryotic cell plasma membranes and, when used at sub-lytic concentrations, significantly augment the cytotoxicity of saporin-based immunotoxins (IT). To help elucidate the mechanism(s) behind these two phenomena we investigated the role of cholesterol to both. Human Daudi lymphoma cells were lipid deprived using a combination of three different approaches. Following treatment, the total cellular lipid content was analyzed by electrospray ionization mass spectrometry (ESI-MS) and plasma membrane (PM) cholesterol content measured using the lipophilic fluorescent probe NR12S. Maximal lipid deprivation of cells resulted in a complete loss of sensitivity to lysis by SA. Similarly augmentation of the anti-CD19 immunotoxin (IT) BU12-SAPORIN by SA was lost but without a concomitant loss of intrinsic IT cytotoxicity. The lytic activity of SA was restored following incubation of lipid deprived Daudi cells with Synthecol or LDL. The augmentative effect of SA on IT cytotoxicity for Daudi cells was restored following repletion of PM cholesterol levels with LDL. NR12S fluorescence and ESI-MS analysis of cellular lipids demonstrated that restoration of SA lytic activity by Synthecol was entirely due to increased PM cholesterol levels. Restoration of cellular and PM cholesterol levels by LDL also restored the augmentative effect of SA for IT, an effect associated with repletion of PM cholesterol with minor changes in some phospholipid species. These results indicate that the lytic and IT augmentative properties of SA are cholesterol-dependent in contrast to intrinsic IT cytotoxicity that is at least partially cholesterol independent.
[Mh] Termos MeSH primário: Antígenos CD19/imunologia
LDL-Colesterol/fisiologia
Imunotoxinas/farmacologia
Linfoma/tratamento farmacológico
Lipídeos de Membrana/fisiologia
Proteínas Inativadoras de Ribossomos Tipo 1/farmacologia
Saponinas/farmacologia
Triterpenos/farmacologia
[Mh] Termos MeSH secundário: Linhagem Celular Tumoral
Seres Humanos
Linfoma/química
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Antigens, CD19); 0 (CD19 molecule, human); 0 (Cholesterol, LDL); 0 (Immunotoxins); 0 (Membrane Lipids); 0 (Ribosome Inactivating Proteins, Type 1); 0 (Saponins); 0 (Triterpenes); EC 3.2.2.22 (saporin)
[Em] Mês de entrada:1708
[Cu] Atualização por classe:170831
[Lr] Data última revisão:
170831
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170226
[St] Status:MEDLINE


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[PMID]:28223166
[Au] Autor:Martinez de Pinillos Bayona A; Woodhams JH; Pye H; Hamoudi RA; Moore CM; MacRobert AJ
[Ad] Endereço:Division of Surgery and Interventional Sciences, University College London, London, United Kingdom. Electronic address: a.martinezdepinillos.12@ucl.ac.uk.
[Ti] Título:Efficacy of photochemical internalisation using disulfonated chlorin and porphyrin photosensitisers: An in vitro study in 2D and 3D prostate cancer models.
[So] Source:Cancer Lett;393:68-75, 2017 May 01.
[Is] ISSN:1872-7980
[Cp] País de publicação:Ireland
[La] Idioma:eng
[Ab] Resumo:This study shows the therapeutic outcome of Photochemical Internalisation (PCI) in prostate cancer in vitro surpasses that of Photodynamic Therapy (PDT) and could improve prostate PDT in the clinic, whilst avoiding chemotherapeutics side effects. In addition, the study assesses the potential of PCI with two different photosensitisers (TPCS and TPPS ) in prostate cancer cells (human PC3 and rat MatLyLu) using standard 2D monolayer culture and 3D biomimetic model. Photosensitisers were used alone for photodynamic therapy (PDT) or with the cytotoxin saporin (PCI). TPPS and TPCS were shown to be located in discrete cytoplasmic vesicles before light treatment and redistribute into the cytosol upon light excitation. PC3 cells exhibit a higher uptake than MatLyLu cells for both photosensitisers. In the 2D model, PCI resulted in greater cell death than PDT alone in both cell lines. In 3D model, morphological changes were also observed. Saporin-based toxicity was negligible in PC3 cells, but pronounced in MatLyLu cells (IC50 = 18 nM). In conclusion, the study showed that tumour features such as tumour cell growth rate or interaction with drugs determine therapeutic conditions for optimal photochemical treatment in metastatic prostate cancer.
[Mh] Termos MeSH primário: Adenocarcinoma/tratamento farmacológico
Benzenossulfonatos/farmacologia
Fotoquimioterapia/métodos
Fármacos Fotossensibilizantes/farmacologia
Porfirinas/farmacologia
Neoplasias da Próstata/tratamento farmacológico
[Mh] Termos MeSH secundário: Adenocarcinoma/metabolismo
Adenocarcinoma/patologia
Animais
Antineoplásicos Fitogênicos/farmacologia
Benzenossulfonatos/metabolismo
Benzenossulfonatos/toxicidade
Transporte Biológico
Materiais Biomiméticos
Linhagem Celular Tumoral
Proliferação Celular/efeitos dos fármacos
Sobrevivência Celular/efeitos dos fármacos
Colágeno/metabolismo
Relação Dose-Resposta a Droga
Seres Humanos
Hidrogéis
Masculino
Fotoquimioterapia/efeitos adversos
Fármacos Fotossensibilizantes/metabolismo
Fármacos Fotossensibilizantes/toxicidade
Porfirinas/metabolismo
Porfirinas/toxicidade
Neoplasias da Próstata/metabolismo
Neoplasias da Próstata/patologia
Ratos
Proteínas Inativadoras de Ribossomos Tipo 1/farmacologia
Fatores de Tempo
[Pt] Tipo de publicação:COMPARATIVE STUDY; JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Antineoplastic Agents, Phytogenic); 0 (Benzenesulfonates); 0 (Hydrogels); 0 (Photosensitizing Agents); 0 (Porphyrins); 0 (Ribosome Inactivating Proteins, Type 1); 0 (TPPS2a chlorin); 0 (meso-tetraphenyl chlorin disulphonate); 2683-84-3 (chlorin); 9007-34-5 (Collagen); EC 3.2.2.22 (saporin)
[Em] Mês de entrada:1708
[Cu] Atualização por classe:170814
[Lr] Data última revisão:
170814
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170223
[St] Status:MEDLINE


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[PMID]:27823883
[Au] Autor:Mason JM; Yuan H; Evans GB; Tyler PC; Du Q; Schramm VL
[Ad] Endereço:The Ferrier Institute, Victoria University of Wellington, 69 Gracefield Rd, Lower Hutt 5010, New Zealand. Electronic address: jennifer.mason@vuw.ac.nz.
[Ti] Título:Oligonucleotide transition state analogues of saporin L3.
[So] Source:Eur J Med Chem;127:793-809, 2017 Feb 15.
[Is] ISSN:1768-3254
[Cp] País de publicação:France
[La] Idioma:eng
[Ab] Resumo:Ribosome inactivating proteins (RIPs) are among the most toxic agents known. More than a dozen clinical trials against refractory cancers have been initiated using modified RIPs with impressive results. However, dose-limiting toxicity due to vascular leak syndrome limits success of the therapy. We have previously reported some tight-binding transition state analogues of Saporin L3 that mimic small oligonucleotide substrates in which the susceptible adenosine has been replaced by a 9-deazaadenyl hydroxypyrrolidinol derivative. They provide the first step in the development of rescue agents to prevent Saporin L3 toxicity on non-targeted cells. Here we report the synthesis, using solution phase chemistry, of these and a larger group of transition state analogues. They were tested for inhibition against Saporin L3 giving K values as low as 3.3 nM and indicating the structural requirements for inhibition.
[Mh] Termos MeSH primário: Materiais Biomiméticos/química
Materiais Biomiméticos/farmacologia
Oligonucleotídeos/química
Oligonucleotídeos/farmacologia
Proteínas Inativadoras de Ribossomos Tipo 1/química
[Mh] Termos MeSH secundário: Sequência de Bases
Modelos Moleculares
Oligonucleotídeos/genética
Conformação Proteica
RNA/genética
RNA/metabolismo
Proteínas Inativadoras de Ribossomos Tipo 1/metabolismo
Proteínas Inativadoras de Ribossomos Tipo 1/toxicidade
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Oligonucleotides); 0 (Ribosome Inactivating Proteins, Type 1); 63231-63-0 (RNA); EC 3.2.2.22 (saporin)
[Em] Mês de entrada:1702
[Cu] Atualização por classe:170425
[Lr] Data última revisão:
170425
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161109
[St] Status:MEDLINE


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[PMID]:27845292
[Au] Autor:Kruashvili L; Demurishvili M; Burjanadze M; Dashniani M; Beselia G
[Ad] Endereço:I. Beritashvili Center of Experimental Biomedicine, Tbilisi, Georgia.
[Ti] Título:EFFECTS OF SELECTIVE CHOLINERGIC AND GABAERGIC LESIONS OF THE NUCLEUS BASALIS MAGNOCELLULARIS ON PLACE OR RESPONCE LEARNING IN PLUS-SHAPED MAZE.
[So] Source:Georgian Med News;(259):77-82, 2016 Oct.
[Is] ISSN:1512-0112
[Cp] País de publicação:Georgia (Republic)
[La] Idioma:eng
[Ab] Resumo:In the present study we evaluated effects of selective cholinergic or GABAergic lesions of the nucleus basalis magnocellularis (NBM) using immunotxins 192 IgG-saporin and GAT1-SAP on place and response learning in plus-shaped maze. In current behavioral paradigm rats learned food-rewarded mazes that were efficiently learned using either place or turning strategies. A histological evaluation indicated that 192 IgG-saporin lesions specifically depleted cholinergic neurons but did not result in noticeable damage to the GABAergic cells within NBM. GAT1-SAP lesions resulted extensive damage of GABAergic and a mild reduction of cholinergic NBM neurons. The results of present behavioral experiments showed, that selective lesions of cholinergic or GABAergic neurons in the NBM impair, but do not abolish, the animal's ability to learn location of rewarded arm of maze (place learning) or a skilled motor behavior (response learning). Our findings suggest the role of NBM cholinergic and GABAergic cortical projection neurons in processing of cognitive information. We suggested that lesions of NBM projections to the cortex modulate learning-mediated plasticity and impair both place and response learning.
[Mh] Termos MeSH primário: Núcleo Basal de Meynert/fisiologia
Neurônios Colinérgicos/fisiologia
Neurônios GABAérgicos/fisiologia
Aprendizagem em Labirinto
Aprendizagem Espacial
[Mh] Termos MeSH secundário: Animais
Anticorpos/toxicidade
Anticorpos Monoclonais/toxicidade
Proteínas da Membrana Plasmática de Transporte de GABA/imunologia
Proteínas da Membrana Plasmática de Transporte de GABA/metabolismo
Imunotoxinas/toxicidade
Masculino
Ratos
Recompensa
Proteínas Inativadoras de Ribossomos Tipo 1/toxicidade
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (192 IgG-saporin); 0 (Antibodies); 0 (Antibodies, Monoclonal); 0 (GABA Plasma Membrane Transport Proteins); 0 (Immunotoxins); 0 (Ribosome Inactivating Proteins, Type 1); EC 3.2.2.22 (saporin)
[Em] Mês de entrada:1703
[Cu] Atualização por classe:170817
[Lr] Data última revisão:
170817
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161116
[St] Status:MEDLINE


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[PMID]:27842546
[Au] Autor:Errico Provenzano A; Posteri R; Giansanti F; Angelucci F; Flavell SU; Flavell DJ; Fabbrini MS; Porro D; Ippoliti R; Ceriotti A; Branduardi P; Vago R
[Ad] Endereço:Istituto Biologia e Biotecnologia Agraria, CNR, Milan, Italy.
[Ti] Título:Optimization of construct design and fermentation strategy for the production of bioactive ATF-SAP, a saporin based anti-tumoral uPAR-targeted chimera.
[So] Source:Microb Cell Fact;15(1):194, 2016 Nov 14.
[Is] ISSN:1475-2859
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:BACKGROUND: The big challenge in any anti-tumor therapeutic approach is represented by the development of drugs selectively acting on the target with limited side effects, that exploit the unique characteristics of malignant cells. The urokinase (urokinase-type plasminogen activator, uPA) and its receptor uPAR have been identified as preferential target candidates since they play a key role in the evolution of neoplasms and are associated with neoplasm aggressiveness and poor clinical outcome in several different tumor types. RESULTS: To selectively target uPAR over-expressing cancer cells, we prepared a set of chimeric proteins (ATF-SAP) formed by the human amino terminal fragments (ATF) of uPA and the plant ribosome inactivating protein saporin (SAP). Codon-usage optimization was used to increase the expression levels of the chimera in the methylotrophic yeast Pichia pastoris. We then moved the bioprocess to bioreactors and demonstrated that the fed-batch production of the recombinant protein can be successfully achieved, obtaining homogeneous discrete batches of the desired constructs. We also determined the cytotoxic activity of the obtained batch of ATF-SAP which was specifically cytotoxic for U937 leukemia cells, while another construct containing a catalytically inactive mutant form of SAP showed no activity. CONCLUSION: Our results demonstrate that the uPAR-targeted, saporin-based recombinant fusion ATF-SAP can be produced in a fed-batch fermentation with full retention of the molecules selective cytotoxicity and hence therapeutic potential.
[Mh] Termos MeSH primário: Proteínas Recombinantes de Fusão/biossíntese
Proteínas Inativadoras de Ribossomos Tipo 1/biossíntese
Ativador de Plasminogênio Tipo Uroquinase/biossíntese
[Mh] Termos MeSH secundário: Reatores Biológicos
Ensaios de Seleção de Medicamentos Antitumorais
Fermentação
Seres Humanos
Pichia/genética
Pichia/metabolismo
Proteínas Recombinantes de Fusão/genética
Proteínas Recombinantes de Fusão/isolamento & purificação
Proteínas Recombinantes de Fusão/farmacologia
Proteínas Inativadoras de Ribossomos Tipo 1/genética
Proteínas Inativadoras de Ribossomos Tipo 1/farmacologia
Células U937
Ativador de Plasminogênio Tipo Uroquinase/genética
Ativador de Plasminogênio Tipo Uroquinase/farmacologia
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Recombinant Fusion Proteins); 0 (Ribosome Inactivating Proteins, Type 1); EC 3.2.2.22 (saporin); EC 3.4.21.73 (Urokinase-Type Plasminogen Activator)
[Em] Mês de entrada:1703
[Cu] Atualização por classe:170330
[Lr] Data última revisão:
170330
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161116
[St] Status:MEDLINE


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[PMID]:27553574
[Au] Autor:Nam H; Kerman IA
[Ad] Endereço:Department of Anatomy and Neurobiology, University of Maryland School of Medicine, United States.
[Ti] Título:A2 noradrenergic neurons regulate forced swim test immobility.
[So] Source:Physiol Behav;165:339-49, 2016 Oct 15.
[Is] ISSN:1873-507X
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:The Wistar-Kyoto (WKY) rat is a widely used animal model of depression, which is characterized by dysregulation of noradrenergic signaling. We previously demonstrated that WKY rats show a unique behavioral profile on the forced swim test (FST), characterized by high levels of immobility upon initial exposure and a greater learning-like response by further increasing immobility upon re-exposure than the genetically related Wistar rats. In the current study we aimed to determine whether altered activation of brainstem noradrenergic cell groups contributes to this behavioral profile. We exposed WKY and Wistar rats, to either 5min of forced swim or to the standard two-day FST (i.e. 15min forced swim on Day 1, followed by 5min on Day 2). We then stained their brains for FOS/tyrosine hydroxylase double-immunocytochemistry to determine potential differences in the activation of the brainstem noradrenergic cell groups. We detected a relative hyperactivation in the locus coeruleus of WKY rats when compared to Wistars in response to both one- and two-day forced swim. In contrast, within the A2 noradrenergic cell group, WKY rats exhibited diminished levels of FOS across both days of the FST, suggesting their lesser activation. We followed up these observations by selectively lesioning the A2 neurons, using anti-dopamine-ß-hydroxylase-conjugated saporin, in Wistar rats, which resulted in increased FST immobility on both days of the test. Together these data indicate that the A2 noradrenergic cell group regulates FST behavior, and that its hypoactivation may contribute to the unique behavioral phenotype of WKY rats.
[Mh] Termos MeSH primário: Neurônios Adrenérgicos/fisiologia
Transtorno Depressivo/patologia
Resposta de Imobilidade Tônica/fisiologia
Natação/psicologia
[Mh] Termos MeSH secundário: Neurônios Adrenérgicos/efeitos dos fármacos
Animais
Modelos Animais de Doenças
Dopamina beta-Hidroxilase/farmacologia
Imunotoxinas/uso terapêutico
Masculino
Microinjeções
Proteínas Oncogênicas v-fos/metabolismo
Ratos
Ratos Endogâmicos WKY
Ratos Sprague-Dawley
Ratos Wistar
Proteínas Inativadoras de Ribossomos Tipo 1/farmacologia
Núcleo Solitário/efeitos dos fármacos
Núcleo Solitário/patologia
Especificidade da Espécie
Fatores de Tempo
Tirosina 3-Mono-Oxigenase/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Immunotoxins); 0 (Oncogene Proteins v-fos); 0 (Ribosome Inactivating Proteins, Type 1); EC 1.14.16.2 (Tyrosine 3-Monooxygenase); EC 1.14.17.1 (Dopamine beta-Hydroxylase); EC 3.2.2.22 (saporin)
[Em] Mês de entrada:1707
[Cu] Atualização por classe:170713
[Lr] Data última revisão:
170713
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:160825
[St] Status:MEDLINE



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