scholarly journals Imaging Sensitive and Drug-Resistant Bacterial Infection with [11C]-TMP: In Vitro and First-in-Human Evaluation

2021 ◽  
Author(s):  
Iris K Lee ◽  
Daniel A Jacome ◽  
Joshua K Cho ◽  
Vincent Tu ◽  
Anthony Young ◽  
...  
Keyword(s):  
Bacterial Infection ◽  
Mammalian Cells ◽  
Bacterial Infections ◽  
The Body ◽  
Response Monitoring ◽  
Drug Resistant ◽  
Bacterial Strains ◽  
Critical Question ◽  
In Vitro Uptake

Recently, several molecular imaging strategies have developed to image bacterial infections in humans. Nuclear approaches, specifically positron emission tomography (PET), affords sensitive detection and the ability to non-invasively locate infections deep within the body. Two key radiotracer classes have arisen: metabolic approaches targeting bacterial specific biochemical transformations, and antibiotic-based approaches that have inherent selectivity for bacteria over mammalian cells. A critical question for clinical application of antibiotic radiotracers is whether resistance to the template antibiotic abrogates specific uptake, thus diminishing the predictive value of the diagnostic test. We recently developed small-molecule PET radiotracers based on the antibiotic trimethoprim (TMP), including [11C]-TMP, and have shown their selectivity for imaging bacteria in preclinical models. Here, we measure the in vitro uptake of [11C]-TMP in pathogenic susceptible and drug-resistant bacterial strains. Both resistant and susceptible bacteria showed similar in vitro uptake, which led us to perform whole genome sequencing of these isolates to identify the mechanisms of TMP resistance that permit retained radiotracer binding. By interrogating these isolate genomes and a broad panel of previously sequenced strains, we reveal mechanisms where uptake or binding of TMP radiotracers can potentially be maintained despite the annotation of genes conferring antimicrobial resistance. Finally, we present several examples of patients with both TMP-sensitive and drug-resistant infections in our first-in-human experience with [11C]-TMP. This work underscores the ability of an antibiotic radiotracer to image bacterial infection in patients, which may allow insights into human bacterial pathogenesis, infection diagnosis, and antimicrobial response monitoring.

scholarly journals Drug resistant gut bacteria mimic a host mechanism for anticancer drug clearance

2019 ◽  
Author(s):  
Peter Spanogiannopoulos ◽  
Patrick H. Bradley ◽  
Jonathan Melamed ◽  
Ysabella Noelle Amora Malig ◽  
Kathy N. Lam ◽  
...  
Keyword(s):  
Mammalian Cells ◽  
Drug Disposition ◽  
Transcriptional Profiling ◽  
Drug Efficacy ◽  
Drug Clearance ◽  
Gut Bacteria ◽  
Resistant Bacteria ◽  
Drug Resistant ◽  
Bacterial Strains

Microbiome surveys indicate that pharmaceuticals are the top predictor of inter-individual variations in gut microbial community structure1, consistent with in vitro evidence that non-antibiotic (i.e. host-targeted) drugs inhibit gut bacterial growth2and are subject to extensive metabolism by the gut microbiome3,4. In oncology, bacterial metabolism has been implicated in both drug efficacy5,6and toxicity7,8; however, the degree to which bacterial sensitivity and metabolism can be driven by conserved pathways also found in mammalian cells remains poorly understood. Here, we show that anticancer fluoropyrimidine drugs broadly inhibit the growth of diverse gut bacterial strains. Media supplementation, transcriptional profiling (RNA-seq), and bacterial genetics implicated pyrimidine metabolism as a key target in bacteria, as in mammalian cells. Drug resistant bacteria metabolized 5FU to its inactive metabolite dihydrofluorouracil (DHFU) mimicking the major host pathway for drug clearance. Functional orthologs of the bacterial operon responsible (preTA) are widespread across human gut bacteria from the Firmicutes and Proteobacteria phyla. The observed conservation of both the targets and pathways for metabolism of therapeutics across domains highlights the need to distinguish the relative contributions of human and microbial cells to drug disposition9, efficacy, and side effect profiles.

scholarly journals Autonomous treatment of bacterial infections in vivo using antimicrobial micro- and nanomachines

2021 ◽  
Author(s):  
Xavier Arque ◽  
Marcelo D. T. Torres ◽  
Tania Patino ◽  
Andreia Boaro ◽  
Samuel Sanchez ◽  
...  
Keyword(s):  
Bacterial Infections ◽  
Target Area ◽  
Drug Resistant ◽  
Infection Site ◽  
Bacterial Strains ◽  
Resistance Development ◽  
Active Motion ◽  
Target Effects

The increasing resistance of bacteria to existing antibiotics constitutes a major public health threat globally. Most current antibiotic treatments are hindered by poor delivery to the infection site, leading to undesired off-target effects and drug resistance development and spread. Here, we describe micro- and nanomachines that effectively and autonomously deliver antibiotic payloads to the target area. The active motion and antimicrobial activity of the silica-based robots are driven by catalysis of the enzyme urease and antimicrobial peptides, respectively. These antimicrobial machines show micromolar bactericidal activity in vitro against different Gram-positive and Gram-negative pathogenic bacterial strains and act by rapidly depolarizing their membrane. Finally, they demonstrated autonomous anti-infective efficacy in vivo in a clinically relevant abscess infection mouse model. In summary, our machines combine navigation, catalytic conversion, and bactericidal capacity to deliver antimicrobial payloads to specific infection sites. This technology represents a much-needed tool to direct therapeutics to their target to help combat drug-resistant infections.

Repurposing of auranofin against bacterial infections: An In silico and In vitro study

2020 ◽  
Vol 16 ◽  
Author(s):  
Nidhi Sharma ◽  
Arti Singh ◽  
Ruchika Sharma ◽  
Anoop Kumar
Keyword(s):  
Broad Spectrum ◽  
In Silico ◽  
Antibacterial Agent ◽  
Bacterial Infections ◽  
In Vitro Assays ◽  
Infection Model ◽  
Synergistic Activity ◽  
Bacterial Strains ◽  
Molecular Docking Study

Aim: The aim of the study was to find out the role of auranofin as a promising broad spectrum antibacterial agent. Methods: In-vitro assays (Percentage growth retardation, Bacterial growth kinetics, Biofilm formation assay) and In-silico study (Molegro virtual docker (MVD) version 6.0 and Molecular operating environment (MOE) version 2008.10 software). Results: The in vitro assays have shown that auranofin has good antibacterial activity against Gram positive and Gram negative bacterial strains. Further, auranofin has shown synergistic activity in combination with ampicillin against S. aureus and B. subtilis whereas in combination with neomycin has just shown additive effect against E. coli, P. aeruginosa and B. pumilus. In vivo results have revealed that auranofin alone and in combination with standard drugs significantly decreased the bioburden in zebrafish infection model as compared to control. The molecular docking study have shown good interaction of auranofin with penicillin binding protein (2Y2M), topoisomerase (3TTZ), UDP-3-O-[3- hydroxymyristoyl] N-acetylglucosaminedeacetylase (3UHM), cell adhesion protein (4QRK), β-lactamase (5CTN) and arylsulphatase (1HDH) enzyme as that of reference ligand which indicate multimodal mechanism of action of auranofin. Finally, MTT assay has shown non-cytotoxic effect of auranofin. Conclusion: In conclusion, auranofin in combination with existing antibiotics could be developed as a broad spectrum antibacterial agent; however, further studies are required to confirm its safety and efficacy. This study provides possibility of use of auranofin apart from its established therapeutic indication in combination with existing antibiotics to tackle the problem of resistance.

Studies on Tapeworm Physiology

1949 ◽  
Vol 26 (1) ◽  
pp. 1-15
Author(s):  
J. D. SMYTH
Keyword(s):  
Bacterial Infections ◽  
Detrimental Effect ◽  
Horse Serum ◽  
Body Cavity ◽  
The Body ◽  
Nutrient Media ◽  
Medium Strength ◽  
By Products ◽  
Histochemical Examination

1. Plerocercoid larvae of the pseudophyllidean cestode Ligula intestinalis from the body cavity of roach, were cultured in vitro at 40°C. in a variety of saline and nutrient media. About 65% of such cultures were aseptic. 2. During cultivation, larvae produced acid by-products (unidentified) and the pH fell rapidly. 3. The presence of these acid by-products slowed down development, or, if present in sufficient quantity, caused death. 4. In order to obtain development in nutrient media in a period (3 days) comparable to that required in a bird (the normal host) it was necessary to renew the medium 24-hourly. 5. 6% of the eggs produced from a worm cultured in horse serum were fertile. Fertile eggs were never obtained from larvae cultured in any other media. 6. Certain bacterial infections had no apparent detrimental effect on development, but others were toxic. 7. Some larvae underwent development in non-nutrient medium (¾ strength Locke's solution). The exact conditions under which this occurred was not determined. 8. Fragments (3 cm. long), of larvae or larvae with either scolex or posterior half removed, underwent development to the stage of oviposition in nutrient media. 9. Histochemical examination revealed that the plerocercoid larvae were almost fat-free. During cultivation, very large quantities of cytoplasmic fat were produced the quantity being proportional to the duration of cultivation. Fat was produced even under starvation conditions (i.e. during cultivation in saline) and can be considered a metabolic by-product. 10. The fresh plerocercoid contained great quantities of glycogen in the parenchyma and muscle regions. After cultivation in nutrient or saline media, considerable quantities were still present.

scholarly journals Pharmacological perspectives from Brazilian Salvia officinalis (Lamiaceae): antioxidant, and antitumor in mammalian cells

2016 ◽  
Vol 88 (1) ◽  
pp. 281-292 ◽  
Author(s):  
CHARLENE S.C. GARCIA ◽  
CAROLINE MENTI ◽  
ANA PAULA F. LAMBERT ◽  
THIAGO BARCELLOS ◽  
SIDNEI MOURA ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Mammalian Cells ◽  
Annexin V ◽  
The Body ◽  
Salvia Officinalis ◽  
Gas Chromatography Mass Spectrometry ◽  
Ionization Mass ◽  
Hek 293 ◽  
Salvia Officinalis L

ABSTRACT Salvia officinalis (Lamiaceae) has been used in south of Brazil as a diary homemade, in food condiment and tea-beverage used for the treatment of several disorders. The objective of this study was to characterize chemical compounds in the hydroalcoholic (ExtHS) and aqueous (ExtAS) extract from Salvia officinalis (L.) by gas chromatography-mass spectrometry (GC-MS) and by high-resolution electrospray ionization mass spectrometry (ESI-QTOF MS/MS), evaluate in vitro ability to scavenge the free radical 2,2-diphenyl-1-picrylhydrazyl (DPPH•) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS•+), catalase (CAT-like) and superoxide dismutase (SOD-like) activity, moreover cytotoxic by MTT assay, alterations on cell morphology by giemsa and apoptotic-induced mechanism for annexin V/propidium iodide. Chemical identification sage extracts revealed the presence of acids and phenolic compounds. In vitro antioxidant analysis for both extracts indicated promising activities. The cytotoxic assays using tumor (Hep-2, HeLa, A-549, HT-29 and A-375) and in non-tumor (HEK-293 and MRC-5), showed selectivity for tumor cell lines. Immunocytochemistry presenting a majority of tumor cells at late stages of the apoptotic process and necrosis. Given the results presented here, Brazilian Salvia officinalis (L.) used as condiment and tea, may protect the body against some disease, in particularly those where oxidative stress is involved, like neurodegenerative disorders, inflammation and cancer.

Recent advances: peptides and self-assembled peptide-nanosystems for antimicrobial therapy and diagnosis

2020 ◽  
Vol 8 (18) ◽  
pp. 4975-4996
Author(s):  
Pengfei Zou ◽  
Wen-Ting Chen ◽  
Tongyi Sun ◽  
Yuanyuan Gao ◽  
Li-Li Li ◽  
...  
Keyword(s):  
Human Health ◽  
Antimicrobial Therapy ◽  
Bacterial Infections ◽  
The Body ◽  
Resistant Bacteria ◽  
Drug Resistant ◽  
Self Assembling ◽  
Recent Advances ◽  
Drug Resistant Bacteria ◽  
Therapy And Diagnosis

Bacterial infections, especially the refractory treatment of drug-resistant bacteria, are one of the greatest threats to human health. Self-assembling peptide-based strategies can specifically detect the bacteria at the site of infection in the body and kill it.

scholarly journals Physics Comes to the Aid of Medicine—Clinically-Relevant Microorganisms through the Eyes of Atomic Force Microscope

Pathogens ◽  
2020 ◽  
Vol 9 (11) ◽  
pp. 969
Author(s):  
Mateusz Cieśluk ◽  
Piotr Deptuła ◽  
Ewelina Piktel ◽  
Krzysztof Fiedoruk ◽  
Łukasz Suprewicz ◽  
...  
Keyword(s):  
Mammalian Cells ◽  
Antimicrobial Agents ◽  
Antimicrobial Treatment ◽  
Point Of View ◽  
Diagnostic Methods ◽  
Drug Resistant ◽  
Bacterial Strains ◽  
Force Microscopy ◽  
Atomic Force ◽  
Recent Developments

Despite the hope that was raised with the implementation of antibiotics to the treatment of infections in medical practice, the initial enthusiasm has substantially faded due to increasing drug resistance in pathogenic microorganisms. Therefore, there is a need for novel analytical and diagnostic methods in order to extend our knowledge regarding the mode of action of the conventional and novel antimicrobial agents from a perspective of single microbial cells as well as their communities growing in infected sites, i.e., biofilms. In recent years, atomic force microscopy (AFM) has been mostly used to study different aspects of the pathophysiology of noninfectious conditions with attempts to characterize morphological and rheological properties of tissues, individual mammalian cells as well as their organelles and extracellular matrix, and cells’ mechanical changes upon exposure to different stimuli. At the same time, an ever-growing number of studies have demonstrated AFM as a valuable approach in studying microorganisms in regard to changes in their morphology and nanomechanical properties, e.g., stiffness in response to antimicrobial treatment or interaction with a substrate as well as the mechanisms behind their virulence. This review summarizes recent developments and the authors’ point of view on AFM-based evaluation of microorganisms’ response to applied antimicrobial treatment within a group of selected bacteria, fungi, and viruses. The AFM potential in development of modern diagnostic and therapeutic methods for combating of infections caused by drug-resistant bacterial strains is also discussed.

scholarly journals Molecular imaging of bacterial infections: Overcoming the barriers to clinical translation

2019 ◽  
Vol 11 (508) ◽  
pp. eaax8251 ◽  
Author(s):  
Alvaro A. Ordonez ◽  
Mark A. Sellmyer ◽  
Gayatri Gowrishankar ◽  
Camilo A. Ruiz-Bedoya ◽  
Elizabeth W. Tucker ◽  
...  
Keyword(s):  
Molecular Imaging ◽  
Early Diagnosis ◽  
Rapid Detection ◽  
Bacterial Infections ◽  
The Body ◽  
Diagnostic Tools ◽  
Drug Resistant ◽  
New Approaches ◽  
Sample Testing ◽  
Direct Sample

Clinical diagnostic tools requiring direct sample testing cannot be applied to infections deep within the body, and clinically available imaging tools lack specificity. New approaches are needed for early diagnosis and monitoring of bacterial infections and rapid detection of drug-resistant organisms. Molecular imaging allows for longitudinal, noninvasive assessments and can provide key information about infectious processes deep within the body.

scholarly journals Evolution of Pseudomonas aeruginosa Virulence as a Result of Phage Predation

2013 ◽  
Vol 79 (19) ◽  
pp. 6110-6116 ◽  
Author(s):  
Zeinab Hosseinidoust ◽  
Theo G. M. van de Ven ◽  
Nathalie Tufenkji
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Mammalian Cells ◽  
Bacterial Infections ◽  
Membrane Damage ◽  
Human Keratinocytes ◽  
Host Population ◽  
Resistant Bacteria ◽  
Content Type ◽  
Metabolic Action

ABSTRACTThe rapid increase in the emergence of antibiotic-resistant bacteria has attracted attention to bacteriophages for treating and preventing bacterial infections. Bacteriophages can drive the diversification ofPseudomonas aeruginosa, giving rise to phage-resistant variants with different phenotypes from their ancestral hosts. In this study, we sought to investigate the effect of phage resistance on cytotoxicity of host populations toward cultured mammalian cells. The library of phage-resistantP. aeruginosaPAO1 variants used was developed previously via experimental evolution of an isogenic host population using phages PP7 and E79. Our results presented herein indicate that the phage-resistant variants developed in a heterogeneous phage environment exhibit a greater ability to impede metabolic action of cultured human keratinocytes and have a greater tendency to cause membrane damage even though they cannot invade the cells in large numbers. They also show a heightened resistance to phagocytosis by model murine macrophages. Furthermore, all isolates produced higher levels of at least one of the secreted virulence factors, namely, total proteases, elastase, phospholipase C, and hemolysins. Reverse transcription-quantitative PCR (RT-qPCR) revealed upregulation in the transcription of a number of genes associated with virulence ofP. aeruginosafor the phage-resistant variants. The results of this study indicate a significant change in thein vitrovirulence ofP. aeruginosafollowing phage predation and highlight the need for caution in the selection and design of phages and phage cocktails for therapeutic use.

Micro- and Nanofluidics for Cell Biology, Cell Therapy, and Cell-Based Drug Testing

2009 ◽  
Author(s):  
Shuichi Takayama ◽  
Dongeun Huh ◽  
Jonathan Song ◽  
Wansik Cha ◽  
Yunseok Heo
Keyword(s):  
Cell Biology ◽  
Mammalian Cells ◽  
Cancer Metastasis ◽  
Dna Analysis ◽  
The Body ◽  
Biological Information ◽  
Screening Methods ◽  
Biological Studies

Many biological studies, drug screening methods, and cellular therapies require culture and manipulation of living cells outside of their natural environment in the body. The gap between the cellular microenvironment in vivo and in vitro, however, poses challenges for obtaining physiologically relevant responses from cells used in basic biological studies or drug screens and for drawing out the maximum functional potential from cells used therapeutically. One of the reasons for this gap is because the fluidic environment of mammalian cells in vivo is microscale and dynamic whereas typical in vitro cultures are macroscopic and static. This presentation will give an overview of efforts in our laboratory to develop microfluidic systems that enable spatio-temporal control of both the chemical and fluid mechanical environment of cells. The technologies and methods close the physiology gap to provide biological information otherwise unobtainable and to enhance cellular performance in therapeutic applications. Specific biomedical topics that will be discussed include, in vitro fertilization on a chip, microfluidic tissue engineering of small airway injuries, breast cancer metastasis on a chip, electrochemical biosensors, and development of tuneable nanofluidic systems towards applications in single molecule DNA analysis.

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