scholarly journals Le diagnostic sérologique de la cowdriose : cinq tests comparés

1993 ◽  
Vol 46 (1-2) ◽  
pp. 123-129
Author(s):  
J.L. Du Plessis ◽  
J.D. Bezuidenhout ◽  
M.S. Brett ◽  
Emmanuel Camus ◽  
Frans Jongejan ◽  
...  
Keyword(s):  
Western Blotting ◽  
Cowdria Ruminantium

Cinq tests sérologiques, l'ELISA indirect, l'ELISA de compétition, deux tests par immunofluorescence indirecte utilisant des antigènes différents, et la technique de Western blotting, ont été comparés sur des sérums de contrôle négatifs ou positifs pour Cowdria ruminantium et des sérums d'animaux de régions indemnes de cowdriose. Aucun des tests ne donnait de réaction positive sur les sérums de contrôle négatifs. En dehors de variations peu importantes dans la sensibilité, il y avait une bonne corrélation entre les 5 tests. Leur spécificité reste contestée, car dans tous les 5 tests, des réactions croisées considérables ont été enregistrées avec des anticorps contre un agent non encore identifié, probablement Ehrlichia.

scholarly journals Diagnostic sérologique de la cowdriose au Zimbabwe. Problèmes et perspectives

1993 ◽  
Vol 46 (1-2) ◽  
pp. 121-121
Author(s):  
Anthony F. Barbet ◽  
N. Tebele ◽  
S. Semu ◽  
T. Peter ◽  
L. Wassink ◽  
...  
Keyword(s):  
Western Blotting ◽  
Grand Nombre ◽  
Cowdria Ruminantium

La valeur potentielle de l'immunoblotting (Western blotting) a été étudiée pour le sérodiagnostic de la cowdriose au Zimbabwe, utilisant des Cowdria ruminantium de culture comme antigène. L'anticorps dominant de la réponse des bovins infectés expérimentalement avec des isolats de C. ruminantium du Zimbabwe était dirigé contre un polypeptide d'environ 32 kDa. Des sérums de bovins et ovins de Floride étaient tous négatifs contre ce polypeptide, et des sérums de régions endémiques pour la cowdriose au Zimbabwe étaient positifs, d'où l'idée d'utiliser cette réaction pour le diagnostic. Toutefois, un grand nombre de réactions positives à l'immunoblot a été obtenu en testant des sérums de bovins et d'ovins de régions du Zimbabwe connues pour être indemnes d'Amblyomma et de cowdriose. La dilution de ces sérums positifs n'a pas permis de les distinguer de sérums positifs en provenance de régions endémiques. Des moutons de régions indemnes, positifs à la réaction, étaient négatifs pour C. ruminantium par PCR, n'étaient pas infectieux pour des tiques, et étaient entièrement sensibles à la cowdriose expérimentale. Il est donc vraisemblable qu'il s'agit de fausses réactions croisées, causées par un organisme apparenté qui existe dans les régions indemnes de cowdriose du Zimbabwe.

scholarly journals La protéine immunodominante de Cowdria ruminantium Cr32 conservée dans le genre Ehrlichia

1993 ◽  
Vol 46 (1-2) ◽  
pp. 145-152
Author(s):  
Frans Jongejan ◽  
N. De Vries ◽  
J. Nieuwenhuijs ◽  
A.H.M. Van Vliet ◽  
L.A. Wassink
Keyword(s):  
Western Blotting ◽  
Cowdria Ruminantium

Les tests sérologiques pour la cowdriose sont perturbés par des réactions croisées dues à des anticorps présents chez des animaux soupçonnés d'être infectés par Ehrlichia spp. On a contrôlé des infections expérimentales par Ehrlichia bovis, E. ovina, E. canis et E. phagocytophila, par l'ELISA de compétition, le western blotting et l'immunofluorescence, utilisant des antigènes de cultures de cellules endothéliales infectées de Cowdria. Les réactions croisées par des anticorps contre Ehrlichia sont attribuables à leur reconnaissance d'épitopes sur la protéine immunodominante de Cowdria, Cr32. Ceci est surtout vrai pour E. canis et E. ovina, beaucoup moins pour E. bovis, mais pas du tout pour E. phagocytophila. De plus, une réaction croisée forte a été démontrée entre Cowdria et des anticorps contre E. chaffeensis. Ces résultats concordent avec les relations phylogénétiques trouvées récemment entre Cowdria et d'autres membres de la tribu des Ehrlichieae par Van Vliet et al. en 1992, qui montrent que Cowdria est étroitement apparentée à E. canis et également à E. chaffeensis. Il est proposé d'étudier des antigènes recombinants de Cowdria pour le développement de tests sérologiques de deuxième génération pour la maladie

Cross-Linked αsγt-Chain Hybrids in Plasma Clots Studied by 1D- and 2D Electrophoresis and Western Blotting

1993 ◽  
Vol 70 (03) ◽  
pp. 438-442 ◽  
Author(s):  
B Grøn ◽  
C Filion-Myklebust ◽  
S Bjørnsen ◽  
P Haidaris ◽  
F Brosstad
Keyword(s):  
Molecular Weight ◽  
Monoclonal Antibodies ◽  
Human Plasma ◽  
Western Blotting ◽  
Isoelectric Point ◽  
Factor Xiii ◽  
Cross Linking ◽  
2D Electrophoresis ◽  
Complex Phenomenon ◽  
Fibrinopeptide A

SummaryFibrinogen and fibrin related chains in reduced human plasma as well as the bonds interlinking partially cross-linked fibrin from plasma clots have been studied by means of 1D- and 2D electrophoresis and Western blotting. Immunovisualization of reduced plasma or partially cross-linked fibrin with monoclonal antibodies specific for the α-chains or the γ-chains have shown that several bands represent material belonging to both chains. In order to decide whether these bands constitute αγ-chain hybrids or superimposed α- and γ-chain dimers, the cross-linked material was separated according to both isoelectric point (pI) and molecular weight (MW) using Pharmacia’s Multiphor II system. Western blotting of the second dimension gels revealed that partially cross-linked fibrin contains αsγt-chain hybrids and γ- polymers, in addition to the well-known γ-dimers and α-polymers. The main αsγt-chain hybrid has a pI between that of the α- and the γ-chains, a MW of about 200 kDa and contains Aα-chains with intact fibrinopeptide A (FPA). It was also observed that soluble fibrinogen/fibrin complexes as well as partially cross-linked fibrin contain degraded α-dimers with MWs close to the γ-dimers. These findings demonstrate that factor XIII-catalyzed cross-linking of fibrin is a more complex phenomenon than earlier recognized.

Plasma Factor V Activation Is Prevented by Activated Protein C in the Presence of Phospholipid Vesicles, Not Platelets

1993 ◽  
Vol 69 (02) ◽  
pp. 124-129 ◽  
Author(s):  
Susan Solymoss ◽  
Kim Thi Phu Nguyen
Keyword(s):  
Western Blotting ◽  
Protein C ◽  
Thrombin Generation ◽  
Blood Platelets ◽  
Activated Protein C ◽  
Phospholipid Vesicles ◽  
Factor V ◽  
Two Stage ◽  
One Stage ◽  
Plasma Factor

SummaryActivated protein C (APC) is a vitamin K dependent anticoagulant which catalyzes the inactivation of factor Va and VIIIa, in a reaction modulated by phospholipid membrane surface, or blood platelets. APC prevents thrombin generation at a much lower concentration when added to recalcified plasma and phospholipid vesicles, than recalcified plasma and platelets. This observation was attributed to a platelet associated APC inhibitor. We have performed serial thrombin, factor V one stage and two stage assays and Western blotting of dilute recalcified plasma containing either phospholipid vesicles or platelets and APC. More thrombin was formed at a given APC concentration with platelets than phospholipid. One stage factor V values increased to higher levels with platelets and APC than phospholipid and APC. Two stage factor V values decreased substantially with platelets and 5 nM APC but remained unchanged with phospholipid and 5 nM APC. Western blotting of plasma factor V confirmed factor V activation in the presence of platelets and APC, but lack of factor V activation with phospholipid and APC. Inclusion of platelets or platelet membrane with phospholipid enhanced rather than inhibited APC catalyzed plasma factor V inactivation. Platelet activation further enhanced factor V activation and inactivation at any given APC concentration.Plasma thrombin generation in the presence of platelets and APC is related to ongoing factor V activation. No inhibition of APC inactivation of FVa occurs in the presence of platelets.

Berberine Alleviates Amyloid-beta Pathogenesis Via Activating LKB1/AMPK Signaling in the Brain of APP/PS1 Transgenic Mice

2019 ◽  
Vol 19 (5) ◽  
pp. 342-348 ◽  
Author(s):  
Zhi-You Cai ◽  
Chuan-Ling Wang ◽  
Tao-Tao Lu ◽  
Wen-Ming Yang
Keyword(s):  
Transgenic Mice ◽  
Western Blotting ◽  
Amyloid Β ◽  
Adenosine Monophosphate ◽  
Camp Response Element Binding ◽  
Enzyme Linked Immunosorbent Assay ◽  
Synaptic Density ◽  
Ampk Signaling ◽  
The Brain ◽  
Post Synaptic Density

Background:Liver kinase B1 (LKB1)/5’-adenosine monophosphate-activated protein kinase (AMPK) signaling, a metabolic checkpoint, plays a neuro-protective role in the pathogenesis of Alzheimer’s disease (AD). Amyloid-β (Aβ) acts as a classical biomarker of AD. The aim of the present study was to explore whether berberine (BBR) activates LKB1/AMPK signaling and ameliorates Aβ pathology.Methods:The Aβ levels were detected using enzyme-linked immunosorbent assay and immunohistochemistry. The following biomarkers were measured by Western blotting: phosphorylated (p-) LKB1 (Ser334 and Thr189), p-AMPK (AMPKα and AMPKβ1), synaptophysin, post-synaptic density protein 95 and p-cAMP-response element binding protein (p-CREB). The glial fibrillary acidic protein (GFAP) was determined using Western blotting and immunohistochemistry.Results:BBR inhibited Aβ expression in the brain of APP/PS1 mice. There was a strong up-regulation of both p-LKB1 (Ser334 and Thr189) and p-AMPK (AMPKα and AMPKβ1) in the brains of APP/PS1 transgenic mice after BBR-treatment (P<0.01). BBR promoted the expression of synaptophysin, post-synaptic density protein 95 and p-CREB(Ser133) in the AD brain, compared with the model mice.Conclusion:BBR alleviates Aβ pathogenesis and rescues synapse damage via activating LKB1/AMPK signaling in the brain of APP/PS1 transgenic mice.

MiR-597 Targeting 14-3-3σ Enhances Cellular Invasion and EMT in Nasopharyngeal Carcinoma Cells

2019 ◽  
Vol 12 (2) ◽  
pp. 105-114 ◽  
Author(s):  
Lisha Xie ◽  
Tao Jiang ◽  
Ailan Cheng ◽  
Ting Zhang ◽  
Pin Huang ◽  
...  
Keyword(s):  
Nasopharyngeal Carcinoma ◽  
Cell Lines ◽  
Western Blotting ◽  
Molecular Mechanisms ◽  
Epithelial Mesenchymal Transition ◽  
Suppressor Gene ◽  
Migration And Invasion ◽  
Mesenchymal Transition ◽  
Downstream Target ◽  
Before And After

Background: Alterations in microRNAs (miRNAs) are related to the occurrence of nasopharyngeal carcinoma (NPC) and play an important role in the molecular mechanism of NPC. Our previous studies show low expression of 14-3-3σ (SFN) is related to the metastasis and differentiation of NPC, but the underlying molecular mechanisms remain unclear. Methods: Through bioinformatics analysis, we find miR-597 is the preferred target miRNA of 14-3-3σ. The expression level of 14-3-3σ in NPC cell lines was detected by Western blotting. The expression of miR-597 in NPC cell lines was detected by qRT-PCR. We transfected miR-597 mimic, miR-597 inhibitor and 14-3-3σ siRNA into 6-10B cells and then verified the expression of 14-3-3σ and EMT related proteins, including E-cadherin, N-cadherin and Vimentin by western blotting. The changes of migration and invasion ability of NPC cell lines before and after transfected were determined by wound healing assay and Transwell assay. Results: miR-597 expression was upregulated in NPC cell lines and repaired in related NPC cell lines, which exhibit a potent tumor-forming effect. After inhibiting the miR-597 expression, its effect on NPC cell line was obviously decreased. Moreover, 14-3-3σ acts as a tumor suppressor gene and its expression in NPC cell lines is negatively correlated with miR-597. Here 14-3-3σ was identified as a downstream target gene of miR-597, and its downregulation by miR-597 drives epithelial-mesenchymal transition (EMT) and promotes the migration and invasion of NPC. Conclusion: Based on these findings, our study will provide theoretical and experimental evidences for molecular targeted therapy of NPC.

scholarly journals Honeysuckle Aqueous Extracts Induced let-7a Suppress EV71 Replication and Pathogenesis In Vitro and In Vivo and Is Predicted to Inhibit SARS-CoV-2

Viruses ◽  
2021 ◽  
Vol 13 (2) ◽  
pp. 308
Author(s):  
Ying-Ray Lee ◽  
Chia-Ming Chang ◽  
Yuan-Chieh Yeh ◽  
Chi-Ying F. Huang ◽  
Feng-Mao Lin ◽  
...  
Keyword(s):  
Protein Expression ◽  
Western Blotting ◽  
Expression Profiles ◽  
Plaque Assay ◽  
Virus Titer ◽  
Enterovirus 71 ◽  
Luciferase Reporter ◽  
Pathogen Invasion

Honeysuckle (Lonicera japonica Thunb) is a traditional Chinese medicine (TCM) with an antipathogenic activity. MicroRNAs (miRNAs) are small non-coding RNA molecules that are ubiquitously expressed in cells. Endogenous miRNA may function as an innate response to block pathogen invasion. The miRNA expression profiles of both mice and humans after the ingestion of honeysuckle were obtained. Fifteen overexpressed miRNAs overlapped and were predicted to be capable of targeting three viruses: dengue virus (DENV), enterovirus 71 (EV71) and SARS-CoV-2. Among them, let-7a was examined to be capable of targeting the EV71 RNA genome by reporter assay and Western blotting. Moreover, honeysuckle-induced let-7a suppression of EV71 RNA and protein expression as well as viral replication were investigated both in vitro and in vivo. We demonstrated that let-7a targeted EV71 at the predicted sequences using luciferase reporter plasmids as well as two infectious replicons (pMP4-y-5 and pTOPO-4643). The suppression of EV71 replication and viral load was demonstrated in two cell lines by luciferase activity, RT-PCR, real-time PCR, Western blotting and plaque assay. Furthermore, EV71-infected suckling mice fed honeysuckle extract or inoculated with let-7a showed decreased clinical scores and a prolonged survival time accompanied with decreased viral RNA, protein expression and virus titer. The ingestion of honeysuckle attenuates EV71 replication and related pathogenesis partially through the upregulation of let-7a expression both in vitro and in vivo. Our previous report and the current findings imply that both honeysuckle and upregulated let-7a can execute a suppressive function against the replication of DENV and EV71. Taken together, this evidence indicates that honeysuckle can induce the expression of let-7a and that this miRNA as well as 11 other miRNAs have great potential to prevent and suppress EV71 replication.

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