scholarly journals Tolerancia in vitro de cultivares de Rubus spp. a estrés hídrico simulado con manitol

2021 ◽  
pp. 46442
Author(s):  
Carlos Millones ◽  
Ernestina Vásquez
Keyword(s):  
Factor B

Introducción. La caracterización de cultivares de Rubus spp. tolerantes al estrés hídrico en campos experimentales es complicado por la dificultad de controlar los factores externos del medio ambiente donde son instalados. La inducción de estrés in vitro es una herramienta eficiente para estudiar los mecanismos de respuesta de las plantas y se emplea en los programas de mejoramiento para la selección de genotipos tolerantes al estrés hídrico. Objetivo. Evaluar la respuesta morfológica y fisiológica in vitro de los explantes en tres cultivares y la accesión silvestre de Rubus spp. bajo condiciones de estrés hídrico simulado con manitol. Materiales y métodos. El estudio se realizó en el Laboratorio de Biología de la Universidad Nacional Toribio Rodríguez de Mendoza de Amazonas, Perú, durante el año 2020. Se empleó un diseño completamente al azar con arreglo factorial (Factor A: cuatro genotipos y Factor B: potenciales hídricos simulados con manitol: 0, -0,2, -0,3 y -0,4 MPa) y cuatro explantes por unidad experimental. Resultados. Los cultivares respondieron de manera distinta bajo estrés hídrico simulado con manitol. Los cultivares Navaho y Tupy registraron mayor tolerancia. Conclusión. Los rasgos morfológicos y fisiológicos relacionados con la longitud de raíz, el contenido hídrico de brote, raíz y hoja permitieron identificar cultivares de Rubus spp. tolerantes al estrés hídrico simulado con manitol en la fase vegetativa.

Schistosoma mansoni: complement activation in human and rodent sera by living parasites of various developmental stages

Parasitology ◽  
1983 ◽  
Vol 87 (1) ◽  
pp. 75-86 ◽  
Author(s):  
A. Ruppel ◽  
U. Rother ◽  
H. Vongerichten ◽  
H. J. Diesfeld
Keyword(s):  
Schistosoma Mansoni ◽  
Complement System ◽  
Developmental Stages ◽  
Alternative Pathway ◽  
Classical Pathway ◽  
Factor B ◽  
The Complement System ◽  
Dose Dependent ◽  
Living Adult

SUMMARYLiving Schistosoma mansoni of various developmental stages were studied with respect to their ability to activate the complement system in sera of humans, mice and rats. Immunofluorescence assays demonstrated that binding of human C3 occurred on fresh schistosomula as well as on schistosomula prepared from mouse lymph-nodes or lungs and on adult schistosomes. However, rodent C3 was deposited only on fresh schistosomula. Deposition of human C3 on the worms' surface required activation of the complement system. The alternative pathway was shown to be involved in deposition of human C3 on schistosomes of all ages, whereas activation of the classical pathway was demonstrable only with fresh schistosomula. Immunoelectrophoretic studies demonstrated a dose-dependent cleavage of human C3 and conversion of factor B by living adult schistosomes. The results demonstrate that the ability of living schistosomes to activate complement in vitro is dependent not only on their developmental stage but also on the species of the serum.

scholarly journals Gluthathione S-transferase-resuscitation-promoting factor B recombinant protein of Mycobacterium tuberculosis induces the production of interferon-γ and interleukin-12 in mice splenocytes

2019 ◽  
Vol 28 (3) ◽  
pp. 234-40
Author(s):  
Andriansjah Rukmana ◽  
Burhanuddin Rasyid ◽  
Fitriyah Sjatha
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Recombinant Protein ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Interferon Γ ◽  
Interleukin 12 ◽  
Potential Candidate ◽  
Factor B ◽  
Latent Tb

BACKGROUND As the only TB vaccine available, Bacillus Calmette-Guérin shows variable efficacy in adults and does not provide protection against the resuscitation of latent TB infections. Resuscitation-promoting factor B (RpfB) is a protein produced by Mycobacterium tuberculosis during the resuscitation phase and is promising as a novel TB vaccine. This study was aimed to analyze the immunogenicity of the gluthathione S-transferase (GST)-RpfB recombinant protein on mice splenocytes in vitro.  METHODS After induction with isopropyl β-D-1-thiogalactopyranoside, the protein was extracted by sonication followed by solubilization in 8 M urea buffer. Protein was then re-natured and purified with a GST chromatography column. The isolated protein was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot using anti-GST antibodies, and its concentration was determined using the Bradford method. Each group of splenocytes was treated with 25 μg/ ml of the recombinant protein (GST-RpfB), GST, and phytohemagglutinin. Antigen induction was repeated twice at 24 and 72 hours. The supernatant was collected at 96 hours and interferon gamma (IFNγ), interleukin (IL-12, IL-4, and IL-10) levels were measured with enzyme-linked immunosorbent assays.  RESULTS GST-RpfB recombinant proteins were expressed in the form of inclusion bodies with a molecular weight of approximately 66 kDa. Based on the independent t-test, GST-RpfB stimulated IFNγ and IL-12 production but not IL-4 and IL-10.  CONCLUSIONS The GST-RpfB protein has been immunogenically proven and is a potential candidate as a novel subunit TB vaccine.

Development Of RNAi Therapeutics Targeting The Complement Pathway

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 2471-2471 ◽  
Author(s):  
Anna Borodovsky ◽  
Kristina Yucius ◽  
Andrew Sprague ◽  
James Butler ◽  
Shannon Fishman ◽  
...  
Keyword(s):  
Cell Lines ◽  
Human Cell ◽  
Hemolytic Activity ◽  
Alternative Pathway ◽  
In Vitro Screening ◽  
Human Cell Lines ◽  
Complement Pathway ◽  
Factor B ◽  
Rnai Therapeutics

Abstract The complement system is a pivotal player in multiple hematological conditions. Antibody blockade of the C5 component of complement has been approved as a treatment for both paroxysmal nocturnal hemoglobinuria (PNH) and atypical hemolytic-uremic syndrome (aHUS), validating C5 as an important therapeutic target. Recently, we developed a robust RNAi therapeutics platform for the delivery of siRNAs to the liver using trivalent GalNAc conjugates, enabling silencing of hepatocyte-expressed genes following subcutaneous (SC) injection. The liver is a major source of C5 and other complement pathway components. The GalNAc conjugate technology allows rapid development of siRNAs targeting multiple members of the complement cascade and evaluation of their silencing in pre-clinical models. To examine the utility of the siRNA approach for targeting complement pathway components we designed and synthesized GalNAc conjugated siRNAs targeting rodent, primate and human C5. Potent siRNA duplexes, showing greater than 95% silencing of C5 mRNA were selected using in vitro screening in human cell lines and mouse primary hepatocytes. C5 silencing and serum hemolytic activity inhibition were evaluated in rodents using single and multi-dose SC treatment regimens. A C5-targeting siRNA conjugate demonstrated a single dose ED50 of 0.625 mg/kg in the mouse with greater than 90% silencing of serum C5 achievable at higher doses. Serum C5 silencing was durable, with recovery starting two weeks after a single SC injection We went on to examine the efficacy of C5 silencing in the rat and observed robust lowering of serum C5 with 2.5 and 5 mg/kg multi-dose regimens, resulting in up to ∼90% inhibition of complement classical pathway hemolytic activity. Evaluation of the translation of this approach to higher species is in progress. Since PNH erythrocyte lysis is thought to be mediated by the activation of the alternative pathway of complement we initiated work on the development of siRNA conjugates targeting Factor B, an essential component of the alternative pathway C3 convertase. siRNAs targeting rodent, primate and human Factor B were identified by in vitro screening and demonstrate >90% silencing of Factor B mRNA in human cell lines and primary mouse hepatocytes. Evaluation of Factor B silencing in rodent models is ongoing. siRNA-mediated silencing of liver-derived complement components is a promising novel therapeutic approach for inhibiting the activity of C5 and other complement pathway targets, with the potential to enable subcutaneous treatment for patients with PNH and related disorders. Disclosures: Borodovsky: Alnylam: Employment. Yucius:Alnylam: Employment. Sprague:Alnylam: Employment. Butler:Alnylam: Employment. Fishman:Alnylam: Employment. Nguyen:Alnylam: Employment. Vaishnaw:Alnylam: Employment. Maier:Alnylam: Employment. Kallanthottathil:Alnylam: Employment. Kuchimanchi:Alnylam: Employment. Manoharan:Alnylam: Employment. Meyers:Alnylam: Employment. Fitzgerald:Alnylam: Employment.

Dobutamine Inhibits Phorbol-Myristate-Acetate-Induced Activation of Nuclear Factor-??B in Human T Lymphocytes In Vitro

2004 ◽  
pp. 1508-1515 ◽  
Author(s):  
Torsten Loop ◽  
Tobias Bross ◽  
Matjaz Humar ◽  
Alexander Hoetzel ◽  
Rene Schmidt ◽  
...  
Keyword(s):  
T Lymphocytes ◽  
Phorbol Myristate Acetate ◽  
Nuclear Factor ◽  
Myristate Acetate ◽  
Factor B ◽  
Human T Lymphocytes

scholarly journals Stomata Index of Cattleya sp. Lindl., Planlet in Drought-Stress Conditions

2019 ◽  
Vol 6 (1) ◽  
pp. 15-19
Author(s):  
Tara Sesafia Paletri ◽  
Endang Nurcahyani ◽  
Yulianty Yulianty ◽  
Rochmah Agustrina
Keyword(s):  
Drought Stress ◽  
Ethylene Glycol ◽  
Water Availability ◽  
Stress Conditions ◽  
Poly Ethylene Glycol ◽  
Factor B ◽  
Peg 6000 ◽  
Poly Ethylene ◽  
Concentration Levels

Cattleya was one of the most popular orchids by Indonesian. Constraints in cultivating Cattleya included the lack of water availability in an area, causing drought stress. Stress in drought in vitro can be simulated by adding Poly Ethylene Glycol 6000 and growth regulators in atonic form. The purpose was to determine the effect of atonic solution and PEG 6000 on the stomata index toward in vitro drought stress conditions. This study was carried out in a 3 x 3 factorial experiment. Factor A was an atonic solution with 3 concentration levels: 0 mL/L, 2 mL/L, and 3 mL/L. Factor B is PEG 6000 with 3 concentration levels: 0% b/v, 20% b/v, 25% b/v. The results showed that the higher the concentration of PEG 6000 and atonic concentrations caused a decrease in the stomata index of Cattleya orchid plantlets under drought stress conditions in vitro.

scholarly journals Anti-Factor B Antibodies and Acute Postinfectious GN in Children

2020 ◽  
Vol 31 (4) ◽  
pp. 829-840 ◽  
Author(s):  
Sophie Chauvet ◽  
Romain Berthaud ◽  
Magali Devriese ◽  
Morgane Mignotet ◽  
Paula Vieira Martins ◽  
...  
Keyword(s):  
Complement Activation ◽  
Alternative Pathway ◽  
Functional Characterization ◽  
C3 Glomerulopathy ◽  
Contributing Factors ◽  
Factor B ◽  
Pathway Activation ◽  
Nephritic Syndrome ◽  
French Registry

BackgroundThe pathophysiology of the leading cause of pediatric acute nephritis, acute postinfectious GN, including mechanisms of the pathognomonic transient complement activation, remains uncertain. It shares clinicopathologic features with C3 glomerulopathy, a complement-mediated glomerulopathy that, unlike acute postinfectious GN, has a poor prognosis.MethodsThis retrospective study investigated mechanisms of complement activation in 34 children with acute postinfectious GN and low C3 level at onset. We screened a panel of anticomplement protein autoantibodies, carried out related functional characterization, and compared results with those of 60 children from the National French Registry who had C3 glomerulopathy and persistent hypocomplementemia.ResultsAll children with acute postinfectious GN had activation of the alternative pathway of the complement system. At onset, autoantibodies targeting factor B (a component of the alternative pathway C3 convertase) were found in a significantly higher proportion of children with the disorder versus children with hypocomplementemic C3 glomerulopathy (31 of 34 [91%] versus 4 of 28 [14%], respectively). In acute postinfectious GN, anti-factor B autoantibodies were transient and correlated with plasma C3 and soluble C5b-9 levels. We demonstrated that anti-factor B antibodies enhance alternative pathway convertase activity in vitro, confirming their pathogenic effect. We also identified crucial antibody binding sites on factor B, including one correlated to disease severity.ConclusionsThese findings elucidate the pathophysiologic mechanisms underlying acute postinfectious GN by identifying anti-factor B autoantibodies as contributing factors in alternative complement pathway activation. At onset of a nephritic syndrome with low C3 level, screening for anti-factor B antibodies might help guide indications for kidney biopsy to avoid misdiagnosed chronic glomerulopathy, such as C3 glomerulopathy, and to help determine therapy.

scholarly journals Development of a Tuberculosis Vaccine Seed: Construction of Resuscitation-Promoting Factor B DNA Vaccine and its Expression in Vitro and in Vivo

2018 ◽  
Vol 22 (1) ◽  
Author(s):  
Andriansjah Rukmana ◽  
Ratih D Saraswati ◽  
Fithriyah Fithriyah ◽  
Aprilia Rakhmawati
Keyword(s):  
Dna Vaccine ◽  
Factor B

scholarly journals Transcription by Methanothermobacter thermautotrophicus RNA Polymerase In Vitro Releases Archaeal Transcription Factor B but Not TATA-Box Binding Protein from the Template DNA

2004 ◽  
Vol 186 (18) ◽  
pp. 6306-6310 ◽  
Author(s):  
Yunwei Xie ◽  
John N. Reeve
Keyword(s):  
Transcription Factor ◽  
Rna Polymerase ◽  
Transcription Initiation ◽  
Tata Box ◽  
Factor B ◽  
Methanothermobacter Thermautotrophicus ◽  
Archaeal Transcription ◽  
Tata Box Binding Protein ◽  
Template Dna

ABSTRACT Transcription initiation in Archaea requires the assembly of a preinitiation complex containing the TATA- box binding protein (TBP), transcription factor B (TFB), and RNA polymerase (RNAP). The results reported establish the fate of Methanothermobacter thermautotrophicus TBP and TFB following transcription initiation by M. thermautotrophicus RNAP in vitro. TFB is released after initiation, during extension of the transcript from 4 to 24 nucleotides, but TBP remains bound to the template DNA. Regulation of archaeal transcription initiation by a repressor competition with TBP for TATA-box region binding must accommodate this observation.

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