Systemic hypoxia enhances exercise-mediated bactericidal and subsequent apoptotic responses in human neutrophils

2009 ◽  
Vol 107 (4) ◽  
pp. 1213-1222 ◽  
Author(s):  
Jong-Shyan Wang ◽  
Ya-Ting Chiu
Keyword(s):  
Bactericidal Activity ◽  
Defense Mechanisms ◽  
Experimental Tests ◽  
Oxidase Inhibitor ◽  
Strenuous Exercise ◽  
Human Neutrophils ◽  
E Coli ◽  
Esherichia Coli ◽  
Diphenylene Iodonium ◽  
Oxidant Production

Phagocytosis and oxidative burst are critical host defense mechanisms in which neutrophils clear invading pathogens. Clearing phagocytic neutrophils by triggering apoptosis is an essential process for controlling inflammation. This study elucidates how various exercise bouts with/without hypoxia affected neutrophil bactericidal activity and subsequent apoptosis in humans. Fifteen sedentary males performed six distinct experimental tests in an air-conditioned normobaric hypoxia chamber: two normoxic exercises [strenuous exercise (SE; up to maximal O2consumption) and moderate exercise (ME; 50% maximal O2consumption for 30 min) while exposed to 21% O2], two hypoxic exercises (ME for 30 min while exposed to 12% and 15% O2), and two hypoxic exposures (resting for 30 min while exposed to 12% and 15% O2). The results showed that 1) plasma complement-C3a desArg/C4a desArg/C5a concentrations were increased, 2) expressions of L-selectin/lymphocyte functin-associated antigen-1/Mac-1/C5aR on neutrophils were enhanced, 3) phagocytosis of neutrophils to Esherichia coli and release of neutrophil oxidant products by E. coli were elevated, and 4) E. coli -induced phosphotidylserine exposure or caspase-3 activation of neutrophils were promoted immediately and 2 h after both 12% O2exposure at rest and with ME as well as normoxic SE. Although neither normoxic ME nor breathing 15% O2at rest influenced these complement- and neutrophil-related immune responses, ME at both 12% and 15% O2resulted in enhanced complement activation in the blood, expressions of opsonic/complement receptors on neutrophils, or the bactericidal activity and apoptosis of neutrophils. Moreover, the increased neutrophil oxidant production and apoptosis by normoxic SE and hypoxic ME were ameliorated by treating neutrophils with diphenylene iodonium (a NADPH oxidase inhibitor). Therefore, we conclude that ME at 12–15% O2enhances bactericidal capacity and facilitates the subsequent apoptosis of neutrophils.

scholarly journals Bactericidal Activity of Human Eosinophilic Granulocytes against Escherichia coli

2001 ◽  
Vol 69 (6) ◽  
pp. 3591-3596 ◽  
Author(s):  
Terese Persson ◽  
Pia Andersson ◽  
Mikael Bodelsson ◽  
Martin Laurell ◽  
Johan Malm ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Bactericidal Activity ◽  
Bacterial Invasion ◽  
No Production ◽  
Minor Importance ◽  
Experimental Conditions ◽  
Bacterial Killing ◽  
Gram Negative ◽  
E Coli ◽  
Diphenylene Iodonium

ABSTRACT Eosinophils participate in allergic inflammation and may have roles in the body's defense against helminthic infestation. Even under noninflammatory conditions, eosinophils are present in the mucosa of the large intestine, where large numbers of gram-negative bacteria reside. Therefore, roles for eosinophils in host defenses against bacterial invasion are possible. In a system for bacterial viable counts, the bactericidal activity of eosinophils and the contribution of different cellular antibacterial systems against Escherichia coli were investigated. Eosinophils showed a rapid and efficient killing of E. coli under aerobic conditions, whereas under anaerobic conditions bacterial killing decreased dramatically. In addition, diphenylene iodonium chloride (DPI), an inhibitor of the NADPH oxidase and thereby of superoxide production, also significantly inhibited bacterial killing. The inhibitor of nitric oxide (NO) productionl-N 5-(1-iminoethyl)-ornithine dihydrochloride did not affect the killing efficiency, suggesting that NO or derivatives thereof are of minor importance under the experimental conditions used. To investigate the involvement of superoxide and eosinophil peroxidase (EPO) in bacterial killing, EPO was blocked by azide. The rate of E. coli killing decreased significantly in the presence of azide, whereas addition of DPI did not further decrease the killing, suggesting that superoxide acts in conjunction with EPO. Bactericidal activity was seen in eosinophil extracts containing granule proteins, indicating that oxygen-independent killing may be of importance as well. The findings suggest that eosinophils can participate in host defense against gram-negative bacterial invasion and that oxygen-dependent killing, i.e., superoxide acting in conjunction with EPO, may be the most important bactericidal effector function of these cells.

scholarly journals Comparison of intracellular bactericidal activities of human neutrophils and eosinophils

Blood ◽  
1978 ◽  
Vol 52 (3) ◽  
pp. 609-617
Author(s):  
LR DeChatelet ◽  
RA Migler ◽  
PS Shirley ◽  
HB Muss ◽  
P Szejda ◽  
...  
Keyword(s):  
Amino Acids ◽  
Bactericidal Activity ◽  
Microscopic Examination ◽  
Human Neutrophils ◽  
Time Intervals ◽  
E Coli ◽  
Direct Microscopic Examination ◽  
Bactericidal Activities ◽  
Number Of Bacteria

Eosinophils from a patient with hypereosinophilia were observed to phagocytize radiolabeled E. coli or S. aureus as effectively as neutrophils from a normal control. This was observed at a number of bacteria/cell ratios and a various time intervals following initial challenge and was confirmed by direct microscopic examination. In spite of comparables rates of phagocytosis, the eosinophils were consistently less capable of killing the bacteria. This correlates with an inability of eosinophil peroxidase to catalyze the peroxidase-H2O2-CI--mediated decarboxylation of amino acids; in contrast, both eosinophil and neutrophil peroxidases showed similar capabilities to iodinate protein in vitro. These data are compatible with the importance of the chloride- mediated mechanism in the bactericidal activity of intact phagocytes.

scholarly journals Biological Trojan Horse: Antigen 43 Provides Specific Bacterial Uptake and Survival in Human Neutrophils

2006 ◽  
Vol 75 (1) ◽  
pp. 30-34 ◽  
Author(s):  
Sara Fexby ◽  
Thomas Bjarnsholt ◽  
Peter Østrup Jensen ◽  
Viktoria Roos ◽  
Niels Høiby ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Human Body ◽  
Defense Mechanisms ◽  
Polymorphonuclear Neutrophils ◽  
Human Neutrophils ◽  
Independent Manner ◽  
E Coli ◽  
Multicellular Aggregates ◽  
Bacterial Uptake ◽  
Antigen 43

ABSTRACT Escherichia coli is a versatile pathogen causing millions of infections in humans every year. This bacterium can form multicellular aggregates when it expresses a self-associating protein, antigen 43 (Ag43), on its surface. We have discovered that Ag43-expressing E. coli cells are efficiently taken up by human defense cells, polymorphonuclear neutrophils (PMNs), in an opsonin-independent manner. Surprisingly, the phagocytosed bacteria were not immediately killed but resided as tight aggregates within the PMNs. Our observations indicate that Ag43-mediated uptake and survival in PMNs constitute a mechanism to subvert one of the primary defense mechanisms of the human body.

scholarly journals Comparison of intracellular bactericidal activities of human neutrophils and eosinophils

Blood ◽  
1978 ◽  
Vol 52 (3) ◽  
pp. 609-617 ◽  
Author(s):  
LR DeChatelet ◽  
RA Migler ◽  
PS Shirley ◽  
HB Muss ◽  
P Szejda ◽  
...  
Keyword(s):  
Amino Acids ◽  
Bactericidal Activity ◽  
Microscopic Examination ◽  
Human Neutrophils ◽  
Time Intervals ◽  
E Coli ◽  
Direct Microscopic Examination ◽  
Bactericidal Activities ◽  
Number Of Bacteria

Abstract Eosinophils from a patient with hypereosinophilia were observed to phagocytize radiolabeled E. coli or S. aureus as effectively as neutrophils from a normal control. This was observed at a number of bacteria/cell ratios and a various time intervals following initial challenge and was confirmed by direct microscopic examination. In spite of comparables rates of phagocytosis, the eosinophils were consistently less capable of killing the bacteria. This correlates with an inability of eosinophil peroxidase to catalyze the peroxidase-H2O2-CI--mediated decarboxylation of amino acids; in contrast, both eosinophil and neutrophil peroxidases showed similar capabilities to iodinate protein in vitro. These data are compatible with the importance of the chloride- mediated mechanism in the bactericidal activity of intact phagocytes.

Viability of Edwardsiella tarda and Esherichia coli preserved with glycerol-tryptone soy broth (TSB) kept at freezing temperature

2013 ◽  
Vol 1 (2) ◽  
pp. 154
Author(s):  
Abdur Rohman ◽  
Frans Ijong ◽  
I K Suwetja
Keyword(s):  
Research And Development ◽  
Germ Plasm ◽  
Freezing Temperature ◽  
Experimental Methods ◽  
Edwardsiella Tarda ◽  
Diagnostic Tools ◽  
Glycerol Concentration ◽  
E Coli ◽  
Esherichia Coli ◽  
Escherchia Coli

Preservation of bacteria carried out in relation to the collection and preservation of germ plasm microbe is useful for research and development or for the establishment of diagnostic tools. Glycerol is a good preservation media but it is not known what doses should be used for effective preservation.  This research used two experimental  methods consisting of 2 factors and 3 treatments. This study aimed to find the best glycerol concentration that can be used to preserve Edwarsiella tarda and Escherchia coli in the -20ºC environment, to understand the viability of bacteria after being preserved and to describe the characteristics of the preserved bacteria. Treatments applied were 10%, 15% and 20%  glycerol in TSB. Viability of the bacteria was analyzed after 7, 14, 28, 35, and 42 days of preservation. Results showed that E.coli bacteria preserved in 15%  glycerol had the highest viability, i.e. 84% and preserved in 10% glycerol had the lowest viability, i.e. 80%. But for E. tarda bacteria preserved in 10% glycerol had the highest viability, i.e. 1.83% and preseved in 15% glycerol had the lowest viability, i.e. 0,55%. Preservasi bakteri dilakukan dalam kaitannya dengan koleksi dan konservasi plasma nutfah mikroba yang berguna untuk penelitian dan pengembangan atau untuk pembentukan alat diagnosa. Gliserol merupakan bahan preservasi yang baik, tetapi belum diketahui dosis yang baik dan efektif untuk preservasi bakteri Edwarsiella tarda dan Escherchia coli pada suhu -20ºC. Penelitian ini dilakukan dengan metode eksperimen yang terdiri dari 2 faktor dan 3 taraf perlakuan, masing-masing perlakuan dengan 3 kali ulangan, media preservasi yang digunakan adalah TSB dan gliserol dengan konsentrasi 10%, 15% dan 20%. Parameter yang diukur adalah viabilitas dan kecocokan/penyimpangan karakteristik biokimia. Penelitian ini dilaksanakan di Laboratorium Balai Karantina Ikan Pengendalian Mutu dan Keamanan Hasil Perikanan Manado, dari bulan September sampai dengan November 2013. Tujuan Penelitian ini adalah untuk menentukan konsentrasi gliserol dalam TSB sebagai media preservasi yang efektif dan efisien pada bakteri  Edwarsiella tarda dan Escherchia coli yang dipreservasi dengan suhu -20ºC dan disimpan selama 42  hari. Hasil penelitian menunjukkan adanya penurunan laju pertumbuhan bakteri selama preservasi. Persentase viabilitas  bakteri E. coli yang tertinggi selama preservasi diperoleh dengan penggunaan gliserol konsentrasi 15% dengan jumlah 84% dan yang terendah adalah dengan penggunaan konsentrasi 10% yakni sebesar 80%, sedangkan untuk E. tarda persentase viabilitas  bakteri yang tertinggi selama preservasi diperoleh dengan penggunaan gliserol konsentrasi 10% dengan jumlah 1,83% dan yang terendah adalah dengan penggunaan konsentrasi 15% yakni sebesar 0,55%. Berdasarkan uji statistik analisis variasi (ANAVA) didapat hasil F hitung E. tarda dan E. coli yang lebih besar  dari FTabel dengan tingkat kepercayaan 95 %.

scholarly journals Magic Peptide: Unique Properties of the LRR11 Peptide in the Activation of Leukotriene Synthesis in Human Neutrophils

2021 ◽  
Vol 22 (5) ◽  
pp. 2671
Author(s):  
Galina M. Viryasova ◽  
Ekaterina A. Golenkina ◽  
Tibor Hianik ◽  
Nataliya V. Soshnikova ◽  
Nina G. Dolinnaya ◽  
...  
Keyword(s):  
Defense Mechanisms ◽  
Specific Binding ◽  
Acoustic Method ◽  
Effective Vaccine ◽  
Human Neutrophils ◽  
Shear Mode ◽  
Vaccine Adjuvants ◽  
High Biological Activity ◽  
Pathogen Elimination ◽  
Cpg Odns

Neutrophil-mediated innate host defense mechanisms include pathogen elimination through bacterial phagocytosis, which activates the 5-lipoxygenase (5-LOX) product synthesis. Here, we studied the effect of synthetic oligodeoxyribonucleotides (ODNs), which mimic the receptor-recognized sites of bacterial (CpG-ODNs) and genomic (G-rich ODNs) DNAs released from the inflammatory area, on the neutrophil functions after cell stimulation with Salmonella typhimurium. A possible mechanism for ODN recognition by Toll-like receptor 9 (TLR9) and RAGE receptor has been proposed. We found for the first time that the combination of the magic peptide LRR11 from the leucine-rich repeat (LRR) of TLR9 with the CpG-ODNs modulates the uptake and signaling from ODNs, in particular, dramatically stimulates 5-LOX pathway. Using thickness shear mode acoustic method, we confirmed the specific binding of CpG-ODNs, but not G-rich ODN, to LRR11. The RAGE receptor has been shown to play an important role in promoting ODN uptake. Thus, FPS-ZM1, a high-affinity RAGE inhibitor, suppresses the synthesis of 5-LOX products and reduces the uptake of ODNs by neutrophils; the inhibitor effect being abolished by the addition of LRR11. The results obtained revealed that the studied peptide-ODN complexes possess high biological activity and can be promising for the development of effective vaccine adjuvants and antimicrobial therapeutics.

scholarly journals Intermittent Hypoxia Affects the Spontaneous DifferentiationIn Vitroof Human Neutrophils into Long-Lived Giant Phagocytes

2016 ◽  
Vol 2016 ◽  
pp. 1-17 ◽  
Author(s):  
Larissa Dyugovskaya ◽  
Slava Berger ◽  
Andrey Polyakov ◽  
Peretz Lavie ◽  
Lena Lavie
Keyword(s):  
Nadph Oxidase ◽  
Intermittent Hypoxia ◽  
Oxidase Inhibitor ◽  
Human Neutrophils ◽  
Hypoxic Conditions ◽  
Oxygen Conditions ◽  
Extended Lifespan ◽  
First Time ◽  
Dose Dependent

Previously we identified, for the first time, a new small-size subset of neutrophil-derived giant phagocytes (Gϕ) which spontaneously developin vitrowithout additional growth factors or cytokines. Gϕare CD66b+/CD63+/MPO+/LC3B+and are characterized by extended lifespan, large phagolysosomes, active phagocytosis, and reactive oxygen species (ROS) production, and autophagy largely controls their formation. Hypoxia, and particularly hypoxia/reoxygenation, is a prominent feature of many pathological processes. Herein we investigated Gϕformation by applying various hypoxic conditions. Chronic intermittent hypoxia (IH) (29 cycles/day for 5 days) completely abolished Gϕformation, while acute IH had dose-dependent effects. Exposure to 24 h (56 IH cycles) decreased their size, yield, phagocytic ability, autophagy, mitophagy, and gp91-phox/p22-phoxexpression, whereas under 24 h sustained hypoxia (SH) the size and expression of LC3B and gp91-phox/p22-phoxresembled Gϕformed in normoxia. Diphenyl iodide (DPI), a NADPH oxidase inhibitor, as well as the PI3K/Akt and autophagy inhibitor LY294002 abolished Gϕformation at all oxygen conditions. However, the potent antioxidant, N-acetylcysteine (NAC) abrogated the effects of IH by inducing large CD66b+/LC3B+Gϕand increased both NADPH oxidase expression and phagocytosis. These findings suggest that NADPH oxidase, autophagy, and the PI3K/Akt pathway are involved in Gϕdevelopment.

DIAGNOSTIC LUNG PUNCTURE IN THE PNEUMONIAS OF INFANTS AND CHILDREN

PEDIATRICS ◽  
1969 ◽  
Vol 44 (4) ◽  
pp. 486-492
Author(s):  
Jerome O. Klein
Keyword(s):  
Defense Mechanisms ◽  
Underlying Disease ◽  
Etiologic Agent ◽  
Upper Respiratory Tract ◽  
E Coli ◽  
Pure Cultures ◽  
Lung Puncture ◽  
Lower Respiratory Disease ◽  
And Staphylococcus Aureus ◽  
Etiologic Diagnosis

Lung punctures were performed on 32 occasions in 28 infants with pneumonia to assist in specific bacteriologic diagnosis. The aspirates yielded pure cultures of Diplococcus pneumoniae and Staphylococcus aureus each in four patients and E. coli in two patients. The procedure is performed as for a thoracentesis and requires no special instruments. Three children had pneumothoraces and one had a small hemoptysis following the procedure, but only one child exhibited even minimal respiratory distress as a result of the tap. The literature on lung aspirates was reviewed with respect to the value and potential liability of the procedure. At present, diagnostic lung punctures should be considered in three groups of children with lower respiratory disease: (1) the critically ill child in whom a specific etiologic diagnosis is of major importance to guide antimicrobial therapy, (2) the child who has deteriorated while on therapy and in whom an etiologic agent is not available from the usual upper respiratory tract cultures, and (3) the child with pneumonia complicated by underlying disease or drugs limiting normal host defense mechanisms. In these three instances, the advantages of a specific etiologic diagnosis outweighs the small risk from the lung puncture.

Effects of Water Source, Dilution, Storage, and Bacterial and Fecal Loads on the Efficacy of Electrolyzed Oxidizing Water for the Control of Escherichia coli O157:H7

2004 ◽  
Vol 67 (7) ◽  
pp. 1377-1383 ◽  
Author(s):  
S. M. L. STEVENSON ◽  
S. R. COOK ◽  
S. J. BACH ◽  
T. A. McALLISTER
Keyword(s):  
Escherichia Coli ◽  
Bactericidal Activity ◽  
Storage Temperature ◽  
Uv Light ◽  
Tap Water ◽  
Organic Matter Content ◽  
Water Source ◽  
Deionized Water ◽  
Escherichia Coli O157 ◽  
E Coli

To evaluate the potential of using electrolyzed oxidizing (EO) water for controlling Escherichia coli O157:H7 in water for livestock, the effects of water source, electrolyte concentration, dilution, storage conditions, and bacterial or fecal load on the oxidative reduction potential (ORP) and bactericidal activity of EO water were investigated. Anode and combined (7:3 anode:cathode, vol/vol) EO waters reduced the pH and increased the ORP of deionized water, whereas cathode EO water increased pH and lowered ORP. Minimum concentrations (vol/vol) of anode and combined EO waters required to kill 104 CFU/ml planktonic suspensions of E. coli O157:H7 strain H4420 were 0.5 and 2.0%, respectively. Cathode EO water did not inhibit H4420 at concentrations up to 16% (vol/vol). Higher concentrations of anode or combined EO water were required to elevate the ORP of irrigation or chlorinated tap water compared with that of deionized water. Addition of feces to EO water products (0.5% anode or 2.0% combined, vol/vol) significantly reduced (P < 0.001) their ORP values to <700 mV in all water types. A relationship between ORP and bactericidal activity of EO water was observed. The dilute EO waters retained the capacity to eliminate a 104 CFU/ml inoculation of E. coli O157:H7 H4420 for at least 70 h regardless of exposure to UV light or storage temperature (4 versus 24°C). At 95 h and beyond, UV exposure reduced ORP, significantly more so (P < 0.05) in open than in closed containers. Bactericidal activity of EO products (anode or combined) was lost in samples in which ORP value had fallen to ≤848 mV. When stored in the dark, the diluted EO waters retained an ORP of >848 mV and bactericidal efficacy for at least 125 h; with refrigeration (4°C), these conditions were retained for at least 180 h. Results suggest that EO water may be an effective means by which to control E. coli O157:H7 in livestock water with low organic matter content.

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