Proliferative enteropathy: a global enteric disease of pigs caused byLawsonia intracellularis

AbstractProliferative enteropathy (PE; ileitis) is a common intestinal disease affecting susceptible pigs raised under various management systems around the world. Major developments in the understanding of PE and its causative agent,Lawsonia intracellularis, have occurred that have led to advances in the detection of this disease and methods to control and prevent it. Diagnostic tools that have improved overall detection and early onset of PE in pigs include various serological and molecular-based assays. Histological tests such as immunohistochemistry continue to be the gold standard in confirmingLawsonia-specific lesions in pigspost mortem. Despite extreme difficulties in isolatingL. intracellularis, innovations in the cultivation and the development of pure culture challenge models, have opened doors to better characterization of the pathogenesis of PE throughin vivoandin vitro L. intracellularis–host interactions. Advancements in molecular research such as the genetic sequencing of the entireLawsoniagenome have provided ways to identify various immunogens, metabolic pathways and methods for understanding the epidemiology of this organism. The determinations of immunological responsiveness in pigs to virulent and attenuated isolates ofL. intracellularisand identification of various immunogens have led to progress in vaccine development.

Download Full-text

Composition and Clinical Significance of Exosomes in Tuberculosis: A Systematic Literature Review

Journal of Clinical Medicine ◽

10.3390/jcm10010145 ◽

2021 ◽

Vol 10 (1) ◽

pp. 145

Author(s):

Fantahun Biadglegne ◽

Brigitte König ◽

Arne C. Rodloff ◽

Anca Dorhoi ◽

Ulrich Sack

Keyword(s):

Vaccine Development ◽

Biological Fluids ◽

Diagnostic Tools ◽

Major Health ◽

Fast Detection ◽

Isolation And Characterization ◽

Culture Supernatants

Tuberculosis (TB) remains a major health issue worldwide. In order to contain TB infections, improved vaccines as well as accurate and reliable diagnostic tools are desirable. Exosomes are employed for the diagnosis of various diseases. At present, research on exosomes in TB is still at the preliminary stage. Recent studies have described isolation and characterization of Mycobacterium tuberculosis (Mtb) derived exosomes in vivo and in vitro. Mtb-derived exosomes (Mtbexo) may be critical for TB pathogenesis by delivering mycobacterial-derived components to the recipient cells. Proteomic and transcriptomic analysis of Mtbexo have revealed a variety of proteins and miRNA, which are utilized by the TB bacteria for pathogenesis. Exosomes have been isolated in body fluids, are amenable for fast detection, and could contribute as diagnostic or prognostic biomarker to disease control. Extraction of exosomes from biological fluids is essential for the exosome research and requires careful standardization for TB. In this review, we summarized the different studies on Mtbexo molecules, including protein and miRNA and the methods used to detect exosomes in biological fluids and cell culture supernatants. Thus, the detection of Mtbexo molecules in biological fluids may have a potential to expedite the diagnosis of TB infection. Moreover, the analysis of Mtbexo may generate new aspects in vaccine development.

Download Full-text

Prediction and characterization of enzymatic activities guided by sequence similarity and genome neighborhood networks

eLife ◽

10.7554/elife.03275 ◽

2014 ◽

Vol 3 ◽

Cited By ~ 56

Author(s):

Suwen Zhao ◽

Ayano Sakai ◽

Xinshuai Zhang ◽

Matthew W Vetting ◽

Ritesh Kumar ◽

...

Keyword(s):

Metabolic Pathways ◽

Large Scale ◽

Sequence Similarity ◽

Gene Clusters ◽

Enzymatic Activities ◽

Or Gene ◽

Enzyme Functions

Metabolic pathways in eubacteria and archaea often are encoded by operons and/or gene clusters (genome neighborhoods) that provide important clues for assignment of both enzyme functions and metabolic pathways. We describe a bioinformatic approach (genome neighborhood network; GNN) that enables large scale prediction of the in vitro enzymatic activities and in vivo physiological functions (metabolic pathways) of uncharacterized enzymes in protein families. We demonstrate the utility of the GNN approach by predicting in vitro activities and in vivo functions in the proline racemase superfamily (PRS; InterPro IPR008794). The predictions were verified by measuring in vitro activities for 51 proteins in 12 families in the PRS that represent ~85% of the sequences; in vitro activities of pathway enzymes, carbon/nitrogen source phenotypes, and/or transcriptomic studies confirmed the predicted pathways. The synergistic use of sequence similarity networks3 and GNNs will facilitate the discovery of the components of novel, uncharacterized metabolic pathways in sequenced genomes.

Download Full-text

In vitro Cultivation and Partial Characterization of Lawsonia intracellularis from a Japanese Field Case of Porcine Proliferative Enteropathy

Journal of Veterinary Medical Science ◽

10.1292/jvms.68.609 ◽

2006 ◽

Vol 68 (6) ◽

pp. 609-613 ◽

Cited By ~ 2

Author(s):

Tomohiro KOYAMA ◽

Takuya HIRAI ◽

Shinya NAGAI

Keyword(s):

Partial Characterization ◽

In Vitro Cultivation ◽

Lawsonia Intracellularis ◽

Field Case ◽

Proliferative Enteropathy

Download Full-text

A Monoclonal Immunoglobulin G Antibody Directed against an Immunodominant Linear Epitope on the Ricin A Chain Confers Systemic and Mucosal Immunity to Ricin

Infection and Immunity ◽

10.1128/iai.00796-09 ◽

2009 ◽

Vol 78 (1) ◽

pp. 552-561 ◽

Cited By ~ 57

Author(s):

Lori M. Neal ◽

Joanne O'Hara ◽

Robert N. Brey ◽

Nicholas J. Mantis

Keyword(s):

Mucosal Immunity ◽

Vaccine Development ◽

Linear Epitope ◽

Serum Antibodies ◽

Polyclonal Igg ◽

A Chain ◽

Fast Acting

ABSTRACT Due to the potential use of ricin and other fast-acting toxins as agents of bioterrorism, there is an urgent need for the development of safe and effective antitoxin vaccines. A candidate ricin subunit vaccine (RiVax) consisting of a recombinant attenuated enzymatic A chain (RTA) has been shown to elicit protective antitoxin antibodies in mice and rabbits and is currently being tested in phase I human clinical trials. However, evaluation of the efficacy of this vaccine for humans is difficult for a number of reasons, including the fact that the key neutralizing B-cell epitopes on RTA have not been fully defined. Castelletti and colleagues (Clin. Exp. Immunol. 136:365-372, 2004) recently identified a linear epitope on RTA, spanning residues L161 to I175, as a primary target of serum antibodies derived from humans who had been treated with ricin immunotoxin. While affinity-purified polyclonal IgG antibodies against this region of RTA were capable of neutralizing ricin in vitro, their capacity to confer protection against ricin challenge in vivo was not determined. In this report, we describe the production and characterization of GD12, a murine monoclonal IgG1 antibody specifically directed against residues 163 to 174 (TLARSFIICIQM) of RTA. GD12 bound ricin holotoxin with high affinity (KD [dissociation constant], 2.9 × 10−9 M) and neutralized it with a 50% inhibitory concentration of ∼0.25 μg/ml, as determined by a Vero cell-based cytotoxicity assay. Passive administration of GD12 was sufficient to protect BALB/c mice against intraperitoneal and intragastric ricin challenges. These data are important in terms of vaccine development, since they firmly establish that preexisting serum antibodies directed against residues 161 to 175 on RTA are sufficient to confer both systemic and mucosal immunity to ricin. The potential of GD12 to serve as a therapeutic following ricin challenge was not explored in this study.

Download Full-text

Characterization of in vitro and in vivo metabolic pathways of the investigational anticancer agent, 2‐methoxyestradiol

Journal of Pharmaceutical Sciences ◽

10.1002/jps.20837 ◽

2007 ◽

Vol 96 (7) ◽

pp. 1821-1831 ◽

Cited By ~ 24

Author(s):

Nehal J. Lakhani ◽

Alex Sparreboom ◽

X.i.a. Xu ◽

Timothy D. Veenstra ◽

Jürgen Venitz ◽

...

Keyword(s):

Metabolic Pathways ◽

Anticancer Agent

Download Full-text

Bacille Calmette-Guérin vaccine reprograms human neonatal lipid metabolism in vitro and in vivo

10.1101/2021.12.03.21267251 ◽

2021 ◽

Author(s):

Joann Diray-Arce ◽

Asimenia Angelidou ◽

Kristoffer Jarlov Jensen ◽

Maria Giulia Conti ◽

Rachel S. Kelly ◽

...

Keyword(s):

Metabolic Pathways ◽

Early Life ◽

Mycobacterial Antigen ◽

Bacille Calmette Guerin ◽

Tlr Agonist ◽

Induced Changes ◽

Insight Into

SummaryVaccines have generally been developed with limited insight into their molecular impact. While systems vaccinology, including metabolomics, enables new characterization of vaccine mechanisms of action, these tools have yet to be applied to infants at high risk of infection and receive the most vaccines. Bacille Calmette-Guérin (BCG) protects infants against disseminated tuberculosis (TB) and TB-unrelated infections via incompletely understood mechanisms. We employed mass spectrometry-based metabolomics of blood plasma to profile BCG-induced infant responses in Guinea Bissau in vivo and the U.S. in vitro. BCG selectively altered plasma lipid pathways, including lysophospholipids. BCG-induced lysophosphatidylcholines (LPCs) correlated with both TLR agonist- and purified protein derivative (PPD, mycobacterial antigen)-induced blood cytokine production in vitro, raising the possibility that LPCs contribute to BCG immunogenicity. Analysis of an independent newborn cohort from The Gambia demonstrated shared vaccine-induced metabolites such as phospholipids and sphingolipids. BCG-induced changes to the plasma lipidome and LPCs may contribute to its immunogenicity and inform the discovery and development of early life vaccines.HighlightsNeonatal BCG immunization generates distinct metabolic shifts in vivo and in vitro across multiple independent cohorts.BCG induces prominent changes in concentrations of plasma lysophospholipids (LPLs)BCG induced changes in plasma lysophosphatidylcholines (LPCs) correlate with BCG effects on TLR agonist- and mycobacterial antigen-induced cytokine responses.Characterization of vaccine-induced changes in metabolism may define predictive signatures of vaccine responses and inform early life vaccine development.Abstract FigureGraphical abstract:BCG vaccination perturbs metabolic pathways in vivo and in vitro.Vaccines have traditionally been developed empirically, with limited insight into their impact on molecular pathways. Metabolomics provides a new approach to characterizing vaccine mechanisms but has not yet been applied to human newborns, who are at the highest risk of infection and receive the most vaccines. Bacille Calmette-Guérin (BCG) prevents disseminated mycobacterial disease in children and can induce broad protection to reduce mortality due to non-TB infections. Underlying mechanisms are incompletely characterized. Employing mass spectrometry-based metabolomics, we demonstrate that early BCG administration alters the human neonatal plasma metabolome, especially lipid metabolic pathways such as lysophosphatidylcholines (LPCs), both in vivo and in vitro. Plasma LPCs correlated with both innate TLR-mediated and PPD antigen-induced cytokine responses suggesting that BCG-induced lipids might contribute to the immunogenicity of this vaccine. Vaccine-induced metabolic changes may provide fresh insights into vaccine immunogenicity and inform the discovery and development of early life vaccines.

Download Full-text

In vivo and in vitro characterization of immediate release dosage forms containing n-acetylcysteine using the dynamic open flow-through test apparatus

10.26226/morressier.57d6b2b7d462b8028d88dd29 ◽

2016 ◽

Author(s):

Maximilian Sager

Keyword(s):

Immediate Release ◽

Dosage Forms ◽

Test Apparatus ◽

Open Flow ◽

In Vitro Characterization ◽

Flow Through

Download Full-text

Rapid, Refined, and Robust Method for Expression, Purification, and Characterization of Recombinant Human Amyloid-beta M1-42

10.26434/chemrxiv.7725002.v1 ◽

2019 ◽

Author(s):

Priya Prakash ◽

Travis Lantz ◽

Krupal P. Jethava ◽

Gaurav Chopra

Keyword(s):

Amyloid Beta ◽

Western Blots ◽

Force Microscopy ◽

E Coli ◽

Atomic Force ◽

Set Up ◽

Short Time

Amyloid plaques found in the brains of Alzheimer’s disease (AD) patients primarily consists of amyloid beta 1-42 (Ab42). Commercially, Ab42 is synthetized using peptide synthesizers. We describe a robust methodology for expression of recombinant human Ab(M1-42) in Rosetta(DE3)pLysS and BL21(DE3)pLysS competent E. coli with refined and rapid analytical purification techniques. The peptide is isolated and purified from the transformed cells using an optimized set-up for reverse-phase HPLC protocol, using commonly available C18 columns, yielding high amounts of peptide (~15-20 mg per 1 L culture) in a short time. The recombinant Ab(M1-42) forms characteristic aggregates similar to synthetic Ab42 aggregates as verified by western blots and atomic force microscopy to warrant future biological use. Our rapid, refined, and robust technique to purify human Ab(M1-42) can be used to synthesize chemical probes for several downstream in vitro and in vivo assays to facilitate AD research.

Download Full-text