Antigenic and genetic characterization of a divergent African virus, Ikoma lyssavirus

In 2009, a novel lyssavirus (subsequently named Ikoma lyssavirus, IKOV) was detected in the brain of an African civet (Civettictis civetta) with clinical rabies in the Serengeti National Park of Tanzania. The degree of nucleotide divergence between the genome of IKOV and those of other lyssaviruses predicted antigenic distinction from, and lack of protection provided by, available rabies vaccines. In addition, the index case was considered likely to be an incidental spillover event, and therefore the true reservoir of IKOV remained to be identified. The advent of sensitive molecular techniques has led to a rapid increase in the discovery of novel viruses. Detecting viral sequence alone, however, only allows for prediction of phenotypic characteristics and not their measurement. In the present study we describe the in vitro and in vivo characterization of IKOV, demonstrating that it is (1) pathogenic by peripheral inoculation in an animal model, (2) antigenically distinct from current rabies vaccine strains and (3) poorly neutralized by sera from humans and animals immunized against rabies. In a laboratory mouse model, no protection was elicited by a licensed rabies vaccine. We also investigated the role of bats as reservoirs of IKOV. We found no evidence for infection among 483 individuals of at least 13 bat species sampled across sites in the Serengeti and Southern Kenya.

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The use of siRNA as a pharmacological tool to assess a role for the transcription factor NF-IL6 in the brain under in vitro and in vivo conditions during LPS-induced inflammatory stimulation

Journal of Basic and Clinical Physiology and Pharmacology ◽

10.1515/jbcpp-2017-0017 ◽

2017 ◽

Vol 28 (6) ◽

Cited By ~ 2

Author(s):

Jelena Damm ◽

Joachim Roth ◽

Rüdiger Gerstberger ◽

Christoph Rummel

Keyword(s):

Transcription Factor ◽

Brain Cells ◽

Nuclear Factor ◽

Si Rna ◽

The Brain ◽

Deficient Mice ◽

Transcription Factor Nuclear Factor

AbstractBackground:Studies with NF-IL6-deficient mice indicate that this transcription factor plays a dual role during systemic inflammation with pro- and anti-inflammatory capacities. Here, we aimed to characterize the role of NF-IL6 specifically within the brain.Methods:In this study, we tested the capacity of short interfering (si) RNA to silence the inflammatory transcription factor nuclear factor-interleukin 6 (NF-IL6) in brain cells underResults:In cells of a mixed neuronal and glial primary culture from the ratConclusions:This approach was, thus, not suitable to characterize the role NF-IL6 in the brain

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Characterization of the role of Fhit in maintenance of genomic integrity following low dose radiation, in vivo and in vitro

10.2172/990617 ◽

2010 ◽

Author(s):

Ya Wang

Keyword(s):

Low Dose ◽

Genomic Integrity ◽

Low Dose Radiation ◽

Dose Radiation

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Tissue-specific characterization of mitochondrial branched-chain keto acid oxidation using a multiplexed assay platform

Biochemical Journal ◽

10.1042/bcj20190182 ◽

2019 ◽

Vol 476 (10) ◽

pp. 1521-1537 ◽

Cited By ~ 4

Author(s):

Emma J. Goldberg ◽

Katherine A. Buddo ◽

Kelsey L. McLaughlin ◽

Regina F. Fernandez ◽

Andrea S. Pereyra ◽

...

Keyword(s):

Acid Oxidation ◽

Valuable Insight ◽

Keto Acid ◽

Isolated Mitochondria ◽

Mouse Tissues ◽

Branched Chain

Abstract Alterations to branched-chain keto acid (BCKA) oxidation have been implicated in a wide variety of human diseases, ranging from diabetes to cancer. Although global shifts in BCKA metabolism—evident by gene transcription, metabolite profiling, and in vivo flux analyses have been documented across various pathological conditions, the underlying biochemical mechanism(s) within the mitochondrion remain largely unknown. In vitro experiments using isolated mitochondria represent a powerful biochemical tool for elucidating the role of the mitochondrion in driving disease. Such analyses have routinely been utilized across disciplines to shed valuable insight into mitochondrial-linked pathologies. That said, few studies have attempted to model in vitro BCKA oxidation in isolated organelles. The impetus for the present study stemmed from the knowledge that complete oxidation of each of the three BCKAs involves a reaction dependent upon bicarbonate and ATP, both of which are not typically included in respiration experiments. Based on this, it was hypothesized that the inclusion of exogenous bicarbonate and stimulation of respiration using physiological shifts in ATP-free energy, rather than excess ADP, would allow for maximal BCKA-supported respiratory flux in isolated mitochondria. This hypothesis was confirmed in mitochondria from several mouse tissues, including heart, liver and skeletal muscle. What follows is a thorough characterization and validation of a novel biochemical tool for investigating BCKA metabolism in isolated mitochondria.

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DR5 Knockout Mice Are Compromised in Radiation-Induced Apoptosis

Molecular and Cellular Biology ◽

10.1128/mcb.25.5.2000-2013.2005 ◽

2005 ◽

Vol 25 (5) ◽

pp. 2000-2013 ◽

Cited By ~ 77

Author(s):

Niklas Finnberg ◽

Joshua J. Gruber ◽

Peiwen Fei ◽

Dorothea Rudolph ◽

Anka Bric ◽

...

Keyword(s):

Cell Death ◽

Null Mouse ◽

Organ Specific ◽

Radiation Induced ◽

Induced Apoptosis ◽

The Brain ◽

Necrosis Factor

ABSTRACT DR5 (also called TRAIL receptor 2 and KILLER) is an apoptosis-inducing membrane receptor for tumor necrosis factor-related apoptosis-inducing ligand (also called TRAIL and Apo2 ligand). DR5 is a transcriptional target of p53, and its overexpression induces cell death in vitro. However, the in vivo biology of DR5 has remained largely unexplored. To better understand the role of DR5 in development and in adult tissues, we have created a knockout mouse lacking DR5. This mouse is viable and develops normally with the exception of having an enlarged thymus. We show that DR5 is not expressed in developing embryos but is present in the decidua and chorion early in development. DR5-null mouse embryo fibroblasts expressing E1A are resistant to treatment with TRAIL, suggesting that DR5 may be the primary proapoptotic receptor for TRAIL in the mouse. When exposed to ionizing radiation, DR5-null tissues exhibit reduced amounts of apoptosis compared to wild-type thymus, spleen, Peyer's patches, and the white matter of the brain. In the ileum, colon, and stomach, DR5 deficiency was associated with a subtle phenotype of radiation-induced cell death. These results indicate that DR5 has a limited role during embryogenesis and early stages of development but plays an organ-specific role in the response to DNA-damaging stimuli.

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In vivo and in vitro response to octreotide LAR in a TSH-secreting adenoma: characterization of somatostatin receptor expression and role of subtype 5

Pituitary ◽

10.1007/s11102-010-0271-2 ◽

2010 ◽

Vol 14 (2) ◽

pp. 141-147 ◽

Cited By ~ 25

Author(s):

Federico Gatto ◽

Federica Barbieri ◽

Lara Castelletti ◽

Marica Arvigo ◽

Alessandra Pattarozzi ◽

...

Keyword(s):

Somatostatin Receptor ◽

Receptor Expression ◽

Octreotide Lar ◽

In Vitro Response

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Identification and Characterization of Two Bordetella avium Gene Products Required for Hemagglutination

Infection and Immunity ◽

10.1128/iai.00140-10 ◽

2010 ◽

Vol 78 (6) ◽

pp. 2370-2376 ◽

Cited By ~ 14

Author(s):

Louise M. Temple ◽

David M. Miyamoto ◽

Manju Mehta ◽

Christian M. Capitini ◽

Stephen Von Stetina ◽

...

Keyword(s):

Whooping Cough ◽

Bioinformatic Analysis ◽

Tracheal Ring ◽

Tracheal Cell ◽

Bordetella Avium ◽

Identification And Characterization

ABSTRACT Bordetella avium causes bordetellosis in birds, a disease similar to whooping cough caused by Bordetella pertussis in children. B. avium agglutinates guinea pig erythrocytes via an unknown mechanism. Loss of hemagglutination ability results in attenuation. We report the use of transposon mutagenesis to identify two genes required for hemagglutination. The genes (hagA and hagB) were adjacent and divergently oriented and had no orthologs in the genomes of other Bordetella species. Construction of in-frame, unmarked mutations in each gene allowed examination of the role of each in conferring erythrocyte agglutination, explanted tracheal cell adherence, and turkey poult tracheal colonization. In all of the in vitro and in vivo assays, the requirement for the trans-acting products of hagA and hagB (HagA and HagB) was readily shown. Western blotting, using antibodies to purified HagA and HagB, revealed proteins of the predicted sizes of HagA and HagB in an outer membrane-enriched fraction. Antiserum to HagB, but not HagA, blocked B. avium erythrocyte agglutination and explanted turkey tracheal ring binding. Bioinformatic analysis indicated the similarity of HagA and HagB to several two-component secretory apparatuses in which one product facilitates the exposition of the other. HagB has the potential to serve as a useful immunogen to protect turkeys against colonization and subsequent disease.

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Computational Insight into the Effect of Natural Compounds on the Destabilization of Preformed Amyloid-β(1-40) Fibrils

10.20944/preprints201805.0380.v1 ◽

2018 ◽

Author(s):

Francesco Tavanti ◽

Alfonso Pedone ◽

Maria Cristina Menziani

Keyword(s):

Therapeutic Strategy ◽

Therapeutic Potential ◽

Structure Stability ◽

Insoluble Form ◽

The Brain ◽

Identification And Characterization ◽

Insight Into

One of the principal hallmarks of Alzheimer’s disease (AD) is related to the aggregation of amyloid-β fibrils in an insoluble form in the brain, also known as amyloidosis. Therefore, a prominent therapeutic strategy against AD consists either in blocking the amyloid aggregation and/or destroying the already formed aggregates. Natural products have shown significant therapeutic potential as amyloid inhibitors from in vitro studies as well as in vivo animal tests. In this study, the interaction of five natural biophenols (curcumin, dopamine, (-)-Epigallocatechin-3-gallate, Quercetin, and Rosmarinic acid) with the amyloid-β(1-40) fibrils has been studied through computational simulations. The results allowed the identification and characterization of the different binding modalities of each compounds and their consequences on fibril dynamics and aggregation. It emerges that the lateral aggregation of the fibrils is strongly influenced by the intercalation of the ligands, which modulate the double-layered structure stability.

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NLRP3- and AIM2-autonomy in a mouse model of MSU crystal-induced acute inflammation in vivo suggests imiquimod-dependent targeting of Il-1[beta] expression as relevant therapy for gout patients.

10.1101/772756 ◽

2019 ◽

Author(s):

Alexandre Mariotte ◽

Aurore Decauwer ◽

Chrystelle Po ◽

Cherine Abou-Faycal ◽

Angelique Pichot ◽

...

Keyword(s):

Mouse Model ◽

Acute Inflammation ◽

Joint Inflammation ◽

Inflammatory Action ◽

Recognition Receptor ◽

Msu Crystals

The role of Monosodium Urate (MSU) crystals in gout pathophysiology is well described, as is the major impact of IL-1b in the inflammatory reaction that constitutes the hallmark of the disease. However, despite the discovery of the NLRP3 inflammasome and its role as a Pattern Recognition Receptor linking the detection of a danger signal (MSU) to IL-1b; secretion in vitro, the precise mechanisms leading to joint inflammation in gout patients are still poorly understood. Here, we provide an extensive clinical, biological and molecular characterization of the acute uratic inflammation mouse model induced by subcutaneous injection of MSU crystals, which accurately mimics human gout. Our work reveals several key features of MSU-dependent inflammation and identifies novel therapeutic opportunities, among which the use of topical application of imiquimod to promote interferon-dependent anti-inflammatory action maybe relevant.

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The Two Faces of Ascorbate: Prooxidant Activity and Radio-Sensitisation

10.26686/wgtn.17014772.v1 ◽

2021 ◽

Author(s):

◽

Georgia Carson

Keyword(s):

Dna Damage ◽

Alternative Therapy ◽

High Dose ◽

Intravenous Injections ◽

Unique Nature ◽

Therapy Option ◽

The Brain

Although not recommended by mainstream oncologists, intravenous injections of pharmacological ascorbate are currently an alternative therapy option for cancer patients. Research has not yet determined whether high-dose ascorbate interacts favourably with radiation therapy to increase DNA damage, and therefore cell death in cancer. Some studies suggest that ascorbate can act as a prooxidant and increase the cytotoxic effect of irradiation in vitro. Glioblastoma multiforme (GBM) is a primary brain astrocytoma that is highly therapy resistant, so patients would be advantaged if ascorbate radiosensitised their cancer. In this investigation, flow cytometry and single cell gel electrophoresis (comet tail assay) were used to measure three indicators of DNA damage in GBM cells in response to ascorbate and irradiation, and were contrasted with immunofluorescence-revealed DNA damage from an intracranial mouse model of GBM. The pro-oxidant, radiosensitisation role of ascorbate was confirmed, as measured by H2AX, 8OHdG, and DSBs in vitro. With all three of these markers of DNA damage, combinations of irradiation and ascorbate had increased damage compared with individual treatments. However preliminary in vivo evidence indicates that increased DNA damage did not occur in an animal model of GBM, and in fact ascorbate may protect from DNA damage in an in vivo context. These findings complement previous results from our lab, and serve to fill in gaps in knowledge specifically around the DNA damaging effects of ascorbate. The unique nature of the brain environment, as enclosed by the blood brain barrier, prevents translation of data from other non-brain cancer studies, as such, this investigation also contributes to the exploration of a much needed avenue of research. Considering the context of ascorbate treatment as a potentially harmful currently used adjuvant, it is imperative to confirm or disprove its efficacy in a clinically relevant environment.

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Characterization of AcMNPV with a deletion of ac69 gene

Microbiology Research ◽

10.4081/mr.2011.e4 ◽

2011 ◽

Vol 2 (1) ◽

pp. 4

Author(s):

Jianhao Ke ◽

Jinwen Wang ◽

Riqiang Deng ◽

Lin Lin ◽

Bei Jinlong ◽

...

Keyword(s):

Spodoptera Frugiperda ◽

Trichoplusia Ni ◽

Virus Production ◽

Viral Infectivity ◽

Methyltransferase Activity ◽

Budded Virus

ORF69 (Ac69) of Autographa californica multiple nucleopolyhedrovirus (AcMNPV) is conserved in some baculovirus genomes. Although it has been shown that Ac69 has cap 0-dependent methyltransferase activity and is not required for budded virus production in Spodoptera frugiperda Sf-9 cells, its role in occlusion-derived virus synthesis and virus oral infectivity is not known. This paper describes generation of an ac69 knockout AcMNPV bacmid mutant and analyses of the influence of ac69 deletion on the viral infectivity in Sf-9 cells and Trichoplusia ni larvae so as to investigate the role of ac69 in the viral life cycle. Results indicated that ac69 deletion has little effect on the production rates and morphogenesis of budded virus and occlusion-derived virus in Sf-9 cells. In addition, animal experiment revealed that the deletion mutant did not affect AcMNPV infectivity for Trichoplusia ni larvae in LD50 and LT50 bioassay when administered orally. These results suggest that ac69 may be dispensable for viral infectivity both in vitro and in vivo.

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