Maintenance of α1-antitrypsin activity by means of co-application of hypochlorous acid-scavengers in vitro and in the supernatant of polymorphonuclear leukocytes

SummaryDefibrotide is a polydeoxyribonucleotide with antithrombotic effects in experimental animal models. Most of the actions of this drug have been observed in in vivo test models but no effects have been reported in in vitro systems. In this paper we demonstrate that defibrotide interferes with polymorphonuclear leukocyte-induced human platelet activation in vitro. This effect was not related to any direct interaction with polymorphonuclear leukocytes or platelets, but was due to the inhibition of cathepsin G, the main biochemical mediator of this cell-cell cooperation. Since cathepsin G not only induces platelet activation but also affects some endothelial cell functions, the anticathepsin G activity of defibrotide could help to explain the antithrombotic effect of this drug.

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The Safety and Antimicrobial Properties of Stabilized Hypochlorous Acid in Acetic Acid Buffer for the Treatment of Acute Wounds—a Human Pilot Study and In Vitro Data

The International Journal of Lower Extremity Wounds ◽

10.1177/15347346211015656 ◽

2021 ◽

pp. 153473462110156

Author(s):

Ewa A. Burian ◽

Lubna Sabah ◽

Klaus Kirketerp-Møller ◽

Elin Ibstedt ◽

Magnus M. Fazli ◽

...

Keyword(s):

Pilot Study ◽

Hypochlorous Acid ◽

Treatment Time ◽

Donor Site ◽

Antimicrobial Properties ◽

Prolonged Treatment ◽

Wound Irrigation ◽

The Mean ◽

And Performance

Acute wounds may require cleansing to reduce the risk of infection. Stabilized hypochlorous acid in acetic buffer (HOCl + buffer) is a novel wound irrigation solution with antimicrobial properties. We performed a first-in-man, prospective, open-label pilot study to document preliminary safety and performance in the treatment of acute wounds. The study enrolled 12 subjects scheduled for a split-skin graft transplantation, where the donor site was used as a model of an acute wound. The treatment time was 75 s, given on 6 occasions. A total of 7 adverse events were regarded as related to the treatment; all registered as pain during the procedure for 2 subjects. One subject had a wound infection at the donor site. The mean colony-forming unit (CFU) decreased by 41% after the treatment, and the mean epithelialization was 96% on both days 14 (standard deviation [SD] 8%) and 21 (SD 10%). The study provides preliminary support for the safety, well-tolerance, and efficacy of HOCl + buffer for acute wounds. The pain was frequent although resolved quickly. Excellent wound healing and satisfying antimicrobial properties were observed. A subsequent in vitro biofilm study also indicated good antimicrobial activity against Pseudomonas aeruginosa with a 96% mean reduction of CFU, when used for a treatment duration of 15 min ( P < .0001), and a 50% decrease for Staphylococcus aureus ( P = .1010). Future larger studies are needed to evaluate the safety and performance of HOCl + buffer in acute wounds, including the promising antimicrobial effect by prolonged treatment on bacterial biofilms.

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The Use of the Vitality and Chemotaxis of Human Polymorphonuclear Leukocytes for the In Vitro Estimation of the Acute Toxicity of the First Ten Chemicals from the MEIC List

Alternatives to Laboratory Animals ◽

10.1177/026119299302100112 ◽

1993 ◽

Vol 21 (1) ◽

pp. 73-80

Author(s):

Matteo Valentino ◽

Francesca Monaco ◽

Maria Antonietta Pizzichini ◽

Mario Governa

Keyword(s):

Ethidium Bromide ◽

Polymorphonuclear Leukocytes ◽

Rank Correlation ◽

Fluorescein Diacetate ◽

Live Cells ◽

In Vitro Toxicity ◽

Ic50 Values ◽

Acute Cytotoxicity ◽

Human Polymorphonuclear Leukocytes

The acute cytotoxicity of the first ten MEIC chemicals has been estimated by others in various cell lines. In the present investigation, isolated human polymorphonuclear leukocytes (PMN) from ten healthy non-smoking laboratory personnel were used to assess in vitro toxicity of the same chemicals. The cells were treated with different concentrations of the respective chemicals for three hours and their vitality and chemotaxis were tested. Vitality was measured by fluorescence microscopy after the addition of fluorescein diacetate and ethidium bromide. Living cells which took up and hydrolysed fluorescein diacetate, and dead cells, stained by ethidium bromide, were counted and the percentage of live cells was calculated. Locomotion stimulated by the chemotactic peptide formyl-methionyl-leucyl-phenylalanine (F-MLP), was measured in blind-well Boyden chambers and a chemotactic index was calculated. The results were mathematically transformed to produce a linear curve, and then fitted by the linear least squares procedure, from which LC50 and IC50 values were obtained by interpolation. All the chemicals decreased the vitality and inhibited the chemotaxis of the PMN. Obviously the chemotactic test was more sensitive than the vitality one. A correlation (r = 0.933) was found between vitality and chemotaxis inhibition. Spearman rank correlation analysis revealed significant correlations between our results and those from in vitro experiments conducted in other laboratories, as well as with data concerning mouse, rat and human lethal doses.

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The Responses of Bioactive Betanin Pigment and Its Derivatives from a Red Beetroot (Beta vulgaris L.) Betalain-Rich Extract to Hypochlorous Acid

International Journal of Molecular Sciences ◽

10.3390/ijms22031155 ◽

2021 ◽

Vol 22 (3) ◽

pp. 1155

Author(s):

Karolina Starzak ◽

Katarzyna Sutor ◽

Tomasz Świergosz ◽

Boris Nemzer ◽

Zbigniew Pietrzkowski ◽

...

Keyword(s):

Beta Vulgaris ◽

Hypochlorous Acid ◽

Innate Immune ◽

Inflammatory Factors ◽

Oxygen Species ◽

Plant Pigments ◽

Important Food ◽

Inflammatory Conditions ◽

Ph Range

Neutrophils produce hypochlorous acid (HOCl) as well as other reactive oxygen species as part of a natural innate immune response in the human body; however, excessive levels of HOCl can ultimately be detrimental to health. Recent reports suggest that betacyanin plant pigments can act as potent scavengers of inflammatory factors and are notably effective against HOCl. Comparison of the in vitro anti-hypochlorite activities of a novel betalain-rich red beetroot (Beta vulgaris L.) extract with its pure betalainic pigments revealed that the extract had the highest anti-hypochlorite activity, far exceeding the activity of all of the betalainic derivatives and selected reference antioxidants. This suggests that it may be an important food-based candidate for management of inflammatory conditions induced by excessive HOCl production. Among all pigments studied, betanidin exhibited the highest activity across the pH range.

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THE EFFECT OF TEMPERATURE ON CHEMILUMINESCENCE BY POLYMORPHONUCLEAR LEUKOCYTES IN VITRO

Biochemical and Clinical Aspects of Oxygen ◽

10.1016/b978-0-12-164380-5.50046-3 ◽

1979 ◽

pp. 725-736 ◽

Cited By ~ 1

Author(s):

John E. Repine ◽

John R. Hoidal ◽

Gregory D. Beall ◽

Fred L. Rasp ◽

Dennis P. Clifford ◽

...

Keyword(s):

Polymorphonuclear Leukocytes ◽

Effect Of Temperature

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POLYMORPHONUCLEAR LEUKOCYTES IN IMMUNOLOGIC REACTIONS

Journal of Experimental Medicine ◽

10.1084/jem.124.4.733 ◽

1966 ◽

Vol 124 (4) ◽

pp. 733-752 ◽

Cited By ~ 184

Author(s):

Charles G. Cochrane ◽

Barbara S. Aikin

Keyword(s):

Basement Membrane ◽

Polymorphonuclear Leukocytes ◽

Electron Microscopic Observation ◽

Severe Damage ◽

Electron Microscopic ◽

Cationic Proteins ◽

Vascular Basement Membrane ◽

Arthus Phenomenon ◽

Immunologic Reactions

Vascular basement membrane was disrupted in the presence of polymorphonuclear leukocytes (PMN's) during two immunologic reactions: The Arthus phenomenon and the reaction to locally injected antibody to vascular basement membrane. This disruption was evidenced by (a) the inability of the basement membrane to retain circulating carbon, by (b) loss of antigenic constituents, and by (c) electron microscopic observation showing actual gaps in the structure of the vascular basement membrane. The factors within PMN's responsible for damage to isolated glomerular basement membrane in vitro were found by isolation procedures to be cathepsins D and E. Cationic proteins of PMN's were separable from the cathepsins. While inducing vascular permeability upon injection, these basic proteins failed to inflict the severe damage to the basement membrane observed in Arthus and antibasement membrane reactions. It is concluded that the full expression of these immunologic lesions requires destruction of the basement membrane possibly brought about by cathepsins D and E. Some of the physicochemical properties of these pathologically active leukocytic factors are given.

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Phagocytosis of polymorphonuclear leukocytes by guinea pig peritoneal macrophages: effects of serum and temperature in vitro

FEMS Microbiology Letters ◽

10.1016/0378-1097(92)90065-v ◽

1992 ◽

Vol 105 (4) ◽

pp. 211-218

Author(s):

Chiyuki Yamamoto

Keyword(s):

Guinea Pig ◽

Polymorphonuclear Leukocytes ◽

Peritoneal Macrophages

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Evaluation of a Novel Therapeutic Approach to Treating Severe Pneumococcal Infection Using a Mouse Model

Clinical and Vaccine Immunology ◽

10.1128/cvi.00120-09 ◽

2009 ◽

Vol 16 (6) ◽

pp. 806-810 ◽

Cited By ~ 7

Author(s):

Nikkol Melnick ◽

Gowrisankar Rajam ◽

George M. Carlone ◽

Jacquelyn S. Sampson ◽

Edwin W. Ades

Keyword(s):

Single Dose ◽

Combination Therapy ◽

Polymorphonuclear Leukocytes ◽

Low Dose ◽

Control Level ◽

Pneumococcal Infection ◽

Amino Acid Peptide ◽

Opsonophagocytic Killing

ABSTRACT P4, a 28-amino-acid peptide, is a eukaryotic cellular activator that enhances specific in vitro opsonophagocytic killing of multiple bacterial pathogens. In a previous study, we successfully recreated this phenomenon in mice in vivo by using a two-dose regimen of P4 and pathogen-specific antibodies, which significantly reduced moribundity in mice. For the present study, we hypothesized that the inclusion of a low-dose antibiotic would make it possible to treat the infected mice with a single dose containing a mixture of P4 and a pathogen-specific antibody. A single dose consisting of P4, intravenous immunoglobulin (IVIG), and ceftriaxone effectively reduced moribundity compared to that of untreated controls (n = 10) by 75% (P < 0.05) and rescued all (10 of 10) infected animals (P < 0.05). If rescued animals were reinfected with Streptococcus pneumoniae and treated with a single dose containing P4, IVIG, and ceftriaxone, they could be rerescued. This observation of the repeated successful use of P4 combination therapy demonstrates a low risk of tolerance development. Additionally, we examined the polymorphonuclear leukocytes (PMN) derived from infected mice and observed that P4 enhanced in vitro opsonophagocytic killing (by >80% over the control level; P < 0.05). This finding supports our hypothesis that PMN are activated by P4 during opsonophagocytosis and the recovery of mice from pneumococcal infection. P4 peptide-based combination therapy may offer an alternative and rapid immunotherapy to treat fulminant pneumococcal infection.

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