scholarly journals Impact of bacterial microcompartment-dependent ethanolamine and propanediol metabolism on Listeria monocytogenes interactions with Caco-2 cells

2021 ◽  
Author(s):  
Zhe Zeng ◽  
Lucas M. Wijnands ◽  
Sjef Boeren ◽  
Eddy J. Smid ◽  
Richard A. Notebaart ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Human Cells ◽  
Cellular Systems ◽  
Anaerobic Growth ◽  
Commensal Microbiota ◽  
Bacterial Microcompartment ◽  
Transmission Electron ◽  
Well Assay ◽  
Phospholipid Degradation

AbstractBacterial microcompartment (BMC) dependent ethanolamine (eut) and propanediol utilization (pdu) has recently been shown to stimulate anaerobic growth of Listeria monocytogenes. This metabolic repertoire conceivably contributes to the competitive fitness of L. monocytogenes in the human gastrointestinal (GI) tract, where these compounds become available following phospholipid degradation and mucus-derived rhamnose metabolism by commensal microbiota. Previous transcriptomics and mutant studies of eut and pdu L. monocytogenes suggested a possible role of eut and pdu BMC metabolism in transmission in foods and pathogenicity, but data on a potential role of L. monocytogenes interaction with human cells is currently absent. First, we ask which cellular systems are expressed in the activation of eut and pdu BMC metabolism and the extent to which these systems are conserved between the states. We find common and unique systems related to metabolic shifts, stress and virulence factors. Next, we hypothesize that these common and unique activated cellular systems contribute to a role in the interaction of L. monocytogenes interaction with human cells. We present evidence that metabolically primed L. monocytogenes with active eut and pdu BMCs, as confirmed by metabolic analysis, transmission electron microscopy and proteomics, show significantly enhanced translocation efficacy compared to non-induced cells in a trans-well assay using Caco-2 cells, while adhesion and invasion capacity was similar. Taken together, our results provide insights into the possible key cellular players that drive translocation efficacy upon eut and pdu BMC activation.

scholarly journals Anaerobic growth of Listeria monocytogenes on rhamnose is stimulated by Vitamin B12 and bacterial microcompartment dependent 1,2-propanediol utilization

2021 ◽  
Author(s):  
Zhe Zeng ◽  
Siming Li ◽  
Sjef Boeren ◽  
Eddy J. Smid ◽  
Richard A. Notebaart ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Vitamin B12 ◽  
Anaerobic Growth ◽  
Physiological Effects ◽  
Human Intestine ◽  
Human Pathogens ◽  
Bacterial Microcompartment ◽  
Food Borne ◽  
Transmission Electron ◽  
Shell Proteins

AbstractThe food-borne pathogen Listeria monocytogenes is able to form proteinaceous organelles called bacterial microcompartments (BMCs) that optimize the utilization of substrates, such as 1,2-propanediol, and confer an anaerobic growth advantage. Rhamnose is a deoxyhexose sugar abundant in a range of environments including the human intestine, and can be degraded in anaerobic conditions into 1,2-propanediol, next to acetate and lactate. Rhamnose-derived 1,2-propanediol has been found to link with BMCs in a limited number of commensal human colonic species and some human pathogens such as Salmonella enterica, but the involvement of BMCs in rhamnose metabolism and potential physiological effects on L. monocytogenes are still unknown. In this study, we firstly test the effect of rhamnose uptake and utilization on anaerobic growth of L. monocytogenes EGDe without and with added vitamin B12, followed by metabolic analysis. We unveil that the vitamin B12-dependent activation of pdu stimulates metabolism and anaerobic growth of L. monocytogenes EGDe on rhamnose via 1,2-propanediol degradation into 1-propanol and propionate. Transmission electron microscopy of pdu-induced cells shows that BMCs are formed and additional proteomics experiments confirm expression of pdu BMC shell proteins and enzymes. Finally, we discuss physiological effects and energy efficiency of L. monocytogenes pdu BMC-driven anaerobic rhamnose metabolism and impact on competitive fitness in environments such as the human intestine.

scholarly journals Anaerobic Growth of Listeria monocytogenes on Rhamnose Is Stimulated by Vitamin B 12 and Bacterial Microcompartment-Dependent 1,2-Propanediol Utilization

mSphere ◽  
2021 ◽  
Author(s):  
Zhe Zeng ◽  
Siming Li ◽  
Sjef Boeren ◽  
Eddy J. Smid ◽  
Richard A. Notebaart ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Molecular Mechanisms ◽  
Foodborne Pathogen ◽  
Anaerobic Growth ◽  
Severe Illness ◽  
Vitamin B ◽  
Anaerobic Conditions ◽  
Content Type ◽  
Bacterial Microcompartment ◽  
Vitamin B 12

Listeria monocytogenes is a foodborne pathogen causing severe illness and, as such, it is crucial to understand the molecular mechanisms contributing to its survival strategy and pathogenicity. Rhamnose is a deoxyhexose sugar abundant in a range of environments, including the human intestine, and can be degraded in anaerobic conditions into 1,2-propanediol.

scholarly journals Bacterial microcompartments linked to the flavin-based extracellular electron transfer drives anaerobic ethanolamine utilization in Listeria monocytogenes

2020 ◽  
Author(s):  
Zhe Zeng ◽  
Sjef Boeren ◽  
Varaang Bhandula ◽  
Samuel H. Light ◽  
Eddy J. Smid ◽  
...  
Keyword(s):  
Electron Transfer ◽  
Listeria Monocytogenes ◽  
Electron Acceptor ◽  
Cell Membranes ◽  
Anaerobic Growth ◽  
Extracellular Electron Transfer ◽  
Severe Illness ◽  
Redox Imbalance ◽  
Bacterial Microcompartments

AbstractEthanolamine (EA) is a valuable microbial carbon and nitrogen source derived from phospholipids present in cell membranes. EA catabolism is suggested to occur in so-called bacterial microcompartments (BMCs) and activation of EA utilization (eut) genes is linked to bacterial pathogenesis. Despite reports showing that activation of eut in Listeria monocytogenes is regulated by a vitamin B12-binding riboswitch and that upregulation of eut genes occurs in mice, it remains unknown whether EA catabolism is BMC dependent. Here, we provide evidence for BMC-dependent anaerobic EA utilization via metabolic analysis, proteomics and electron microscopy. First, we show B12-induced activation of the eut operon in L. monocytogenes coupled to uptake and utilization of EA thereby enabling growth. Next, we demonstrate BMC formation in conjunction to EA catabolism with the production of acetate and ethanol in a molar ratio of 2:1. Flux via the ATP generating acetate branch causes an apparent redox imbalance due to reduced regeneration of NAD+ in the ethanol branch resulting in a surplus of NADH. We hypothesize that the redox imbalance is compensated by linking eut BMC to anaerobic flavin-based extracellular electron transfer (EET). Using L. monocytogenes wild type, a BMC mutant and a EET mutant, we demonstrate an interaction between BMC and EET and provide evidence for a role of Fe3+ as an electron acceptor. Taken together, our results suggest an important role of anaerobic BMC-dependent EA catabolism in the physiology of L. monocytogenes, with a crucial role for the flavin-based EET system in redox balancing.IMPORTANCEListeria monocytogenes is a food-borne pathogen causing severe illness and, as such, it is crucial to understand the molecular mechanisms contributing to pathogenicity. One carbon source that allows L. monocytogenes to grow in humans is ethanolamine (EA), which is derived from phospholipids present in eukaryotic cell membranes. It is hypothesized that EA utilization occurs in bacterial microcompartments (BMCs), self-assembling subcellular proteinaceous structures and analogs of eukaryotic organelles. Here, we demonstrate that BMC-driven utilization of EA in L. monocytogenes results in increased energy production essential for anaerobic growth. However, exploiting BMCs and the encapsulated metabolic pathways also requires balancing of oxidative and reductive pathways. We now provide evidence that L. monocytogenes copes with this by linking BMC activity to flavin-based extracellular electron transfer (EET) using iron as an electron acceptor. Our results shed new light on an important molecular mechanism that enables L. monocytogenes to grow using host-derived phospholipid degradation products.

scholarly journals Impact of vitamin B12 on rhamnose metabolism, stress defense and in-vitro virulence of Listeria monocytogenes

2021 ◽  
Author(s):  
Zhe Zeng ◽  
Lucas M. Wijnands ◽  
Sjef Boeren ◽  
Eddy J. Smid ◽  
Richard A. Notebaart ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Vitamin B12 ◽  
Growth Stimulation ◽  
Anaerobic Growth ◽  
Salmonella Spp ◽  
Defense Proteins ◽  
Bacterial Microcompartment ◽  
The Impact ◽  
Stress Defense

Listeria monocytogenes is a facultative anaerobe which can cause a severe food-borne infection known as listeriosis. Rhamnose is a deoxyhexose sugar abundant in a range of environments, including the human intestine, and can be degraded by L. monocytogenes in aerobic and anaerobic conditions into lactate, acetate and 1,2-propanediol. Our previous study showed that addition of vitamin B12 stimulates anaerobic growth of L. monocytogenes on rhamnose due to the activation of bacterial microcompartment (BMC)-dependent 1,2-propanediol utilization with concomitant production of propionate and propanol. Notably, anaerobic propanediol metabolism has been linked to virulence of enteric pathogens including Salmonella spp. and L. monocytogenes. In this study we investigate the impact of B12 on aerobic and anerobic growth of L. monocytogenes on rhamnose, and observed growth stimulation and pdu BMC activation only in anaerobically grown cells with B12 added to the medium. Comparative Caco-2 virulence assays, showed that these pdu BMC induced cells have significantly higher translocation efficiency compared to aerobically grown cells (without and with added B12) and non-induced anaerobically grown cells, while adhesion and invasion capacity is similar for all cells. Comparative proteomics analysis showed specific and overlapping responses linked to metabolic shifts, activation of stress defense proteins and virulence factors, with RNA polymerase sigma factor SigL; teichoic acids export ATP-binding protein, TagH; DNA repair and protection proteins RadA and DPS; and glutathione synthase GshAB previously linked to activation of virulence response in L. monocytogenes, uniquely upregulated in anaerobically rhamnose grown pdu BMC induced cells. Our results shed new light into B12 impact on L. monocytogenes competitive fitness and virulence.

On Future Directions in Pathology

1982 ◽  
Vol 40 ◽  
pp. 274-277
Author(s):  
Benjamin F. Trump ◽  
Irene K. Berezesky ◽  
Raymond T. Jones
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Clinical Pathology ◽  
Future Directions ◽  
Diagnostic Pathology ◽  
The Past ◽  
Transmission Electron ◽  
Organ Systems ◽  
The Many

The role of electron microscopy and associated techniques is assured in diagnostic pathology. At the present time, most of the progress has been made on tissues examined by transmission electron microscopy (TEM) and correlated with light microscopy (LM) and by cytochemistry using both plastic and paraffin-embedded materials. As mentioned elsewhere in this symposium, this has revolutionized many fields of pathology including diagnostic, anatomic and clinical pathology. It began with the kidney; however, it has now been extended to most other organ systems and to tumor diagnosis in general. The results of the past few years tend to indicate the future directions and needs of this expanding field. Now, in addition to routine EM, pathologists have access to the many newly developed methods and instruments mentioned below which should aid considerably not only in diagnostic pathology but in investigative pathology as well.

Transmission Electron Microscopy studies of texture of Cr underlayer of magnetic recording hard disk

1991 ◽  
Vol 49 ◽  
pp. 580-581
Author(s):  
L. Tang ◽  
G. Thomas ◽  
M. R. Khan ◽  
S. L. Duan
Keyword(s):  
Electron Microscopy ◽  
Thin Films ◽  
Transmission Electron Microscopy ◽  
Magnetic Recording ◽  
Magnetic Thin Films ◽  
Magnetic Films ◽  
Substrate Bias ◽  
Epitaxial Relationship ◽  
Transmission Electron

Cr thin films are often used as underlayers for Co alloy magnetic thin films, such as Co1, CoNi2, and CoNiCr3, for high density longitudinal magnetic recording. It is belived that the role of the Cr underlayer is to control the growth and texture of the Co alloy magnetic thin films, and, then, to increase the in plane coercivity of the films. Although many epitaxial relationship between the Cr underlayer and the magnetic films, such as ﹛1010﹜Co/ {110﹜Cr4, ﹛2110﹜Co/ ﹛001﹜Cr5, ﹛0002﹜Co/﹛110﹜Cr6, have been suggested and appear to be related to the Cr thickness, the texture of the Cr underlayer itself is still not understood very well. In this study, the texture of a 2000 Å thick Cr underlayer on Nip/Al substrate for thin films of (Co75Ni25)1-xTix dc-sputtered with - 200 V substrate bias is investigated by electron microscopy.

Transmission Electron Microscopy of Evaporated Perylene Thin Films

1990 ◽  
Vol 48 (2) ◽  
pp. 336-337
Author(s):  
C. Ewins ◽  
J.R. Fryer
Keyword(s):  
Electron Microscopy ◽  
Thin Films ◽  
Transmission Electron Microscopy ◽  
Molecular Electronics ◽  
High Vacuum ◽  
Amorphous Layer ◽  
Electric Heater ◽  
Transmission Electron ◽  
Polycyclic Aromatic

The preparation of thin films of organic molecules is currently receiving much attention because of the need to produce good quality thin films for molecular electronics. We have produced thin films of the polycyclic aromatic, perylene C10H12 by evaporation under high vacuum onto a potassium chloride (KCl) substrate. The role of substrate temperature in determining the morphology and crystallography of the films was then investigated by transmission electron microscopy (TEM).The substrate studied was the (001) face of a freshly cleaved crystal of KCl. The temperature of the KCl was controlled by an electric heater or a cold finger. The KCl was heated to 200°C under a vacuum of 10-6 torr and allowed to cool to the desired temperature. The perylene was then evaporated over a period of one minute from a molybdenum boat at a distance of 10cm from the KCl. The perylene thin film was then backed with an amorphous layer of carbon and floated onto copper microscope grids.

Fine structure and possible functions of the hermaphrodite duct of some pulmonate snails (Mollusca: Gastropoda)

1990 ◽  
Vol 48 (3) ◽  
pp. 68-69
Author(s):  
Alan N. Hodgson
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Fine Structure ◽  
Seminal Vesicle ◽  
Reproductive Biology ◽  
Light Microscope ◽  
Light Microscope Level ◽  
Transmission Electron ◽  
Standard Techniques

The hermaphrodite duct of pulmonate snails connects the ovotestis to the fertilization pouch. The duct is typically divided into three zones; aproximal duct which leaves the ovotestis, the middle duct (seminal vesicle) and the distal ovotestis duct. The seminal vesicle forms the major portion of the duct and is thought to store sperm prior to copulation. In addition the duct may also play a role in sperm maturation and degredation. Although the structure of the seminal vesicle has been described for a number of snails at the light microscope level there appear to be only two descriptions of the ultrastructure of this tissue. Clearly if the role of the hermaphrodite duct in the reproductive biology of pulmonatesis to be understood, knowledge of its fine structure is required.Hermaphrodite ducts, both containing and lacking sperm, of species of the terrestrial pulmonate genera Sphincterochila, Levantina, and Helix and the marine pulmonate genus Siphonaria were prepared for transmission electron microscopy by standard techniques.

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