scholarly journals Complex coacervation-based loading and tunable release of a cationic protein from monodisperse glycosaminoglycan microgels

Soft Matter ◽  
2018 ◽  
Vol 14 (30) ◽  
pp. 6327-6341 ◽  
Author(s):  
Carl C. L. Schuurmans ◽  
Anna Abbadessa ◽  
Mikkel A. Bengtson ◽  
Galja Pletikapic ◽  
Huseyin Burak Eral ◽  
...  
Keyword(s):  
Biomedical Applications ◽  
Complex Coacervation ◽  
Cationic Protein ◽  
Cationic Proteins ◽  
Tunable Release

Glycosaminoglycan-based microgels are of interest for biomedical applications because of their ability to retain and gradually release bioactive cationic proteins.

Ultrastructural localization of cationic proteins in cytoplasmic granules of chicken and rabbit polymorphonuclear leukocytes

1975 ◽  
Vol 17 (1) ◽  
pp. 79-94
Author(s):  
E.K. Macrae ◽  
J.K. Spitznagel
Keyword(s):  
Polymorphonuclear Leukocytes ◽  
Intracellular Localization ◽  
Ultrastructural Localization ◽  
Cationic Protein ◽  
Cationic Proteins ◽  
Cytoplasmic Granules ◽  
Electron Dense Deposit ◽  
Dense Deposit ◽  
Neutrophilic Polymorphonuclear Leukocytes ◽  
Ammoniacal Silver

Cytoplasmic granules known to contain cationic arginine-rich proteins can be identified by the ammoniacal silver reaction (ASR) which provides a cytochemical marker detectable under the electron microscope. Only the large rod-shaped granules of the chicken polymorphonuclear leukocytes (heterophils) and the large spherical azurophilic granules of the rabbit neutrophilic polymorphonuclear leukocytes show the ASR product as a discrete particulate electron-dense deposit. The other smaller granules are devoid of reaction product, as are membranes and mitochondria. The intracellular localization of the ASR product, as are membranes and mitochondria. The intracellular localization of the ASR product on the large granules coincides with the ASR product localization on the same isolated granule populations, when the ammoniacal silver reaction is applied to these granules after their separation by sucrose-density gradients. The cationic proteins may have intraleukocytic bacteriolytic properties, since ASR product, presumably indicating cationic protein from discharged granules, appears to surround ingested bacteria within cytoplasmic phagosomes.

scholarly journals Cationic Proteins of Human Granulocytes. II. Separation of the Cationic Proteins of the Granules of Leukemic Myeloid Cells

Blood ◽  
1974 ◽  
Vol 44 (2) ◽  
pp. 235-246 ◽  
Author(s):  
I. Olsson ◽  
P. Venge
Keyword(s):  
Amino Acid ◽  
Amino Acid Composition ◽  
Acid Composition ◽  
Aminocaproic Acid ◽  
Cationic Protein ◽  
Cationic Proteins ◽  
Molecular Weights ◽  
Electrophoretic Mobilities ◽  
Human Granulocytes ◽  
Protein Components

Abstract The highly cationic proteins of human granulocytes, whose electrophoretic mobilities toward the cathode are faster than that for lysozyme, were isolated from the cytoplasmic granules of leukocytes, obtained from patients with chronic myeloid leukemia. The granule extract was subjected to chromatography on Sephadex G-75 and E-aminocaproic acid-Sepharose ion adsorbant followed by preparative electrophoresis on agarose. Seven cationic protein components were identified, and five of these were obtained in a pure form. One group of cationic proteins, including components 1-4, exhibited molecular weights in the range 25,500-28,500, almost identical amino acid composition, and complete immunologic identity. Another group of proteins, including components 5-7, exhibited molecular weights in the range 21,000-29,000 and also showed complete immunologic identity; amino acid analysis performed on component 5 indicated a different amino acid composition from that of components 1-4. Cationic proteins with similar electrophoretic mobilities and immunochemical identities were also detected in granule extracts of granulocytes from healthy individuals. The proteins isolated from human granulocytes have a higher molecular weight and a lower content of basic amino acids than the cationic proteins with antibacterial and permeability-increasing properties previously demonstrated in rabbit polymorphonuclear granulocytes.

scholarly journals Effects of human eosinophil granule-derived cationic proteins on C-fiber afferents in the rat lung

2001 ◽  
Vol 91 (3) ◽  
pp. 1318-1326 ◽  
Author(s):  
Lu-Yuan Lee ◽  
Qihai Gu ◽  
Gerald J. Gleich
Keyword(s):  
Eosinophil Cationic Protein ◽  
Eosinophil Infiltration ◽  
Cationic Protein ◽  
Human Eosinophil ◽  
Lung Inflation ◽  
Cationic Proteins ◽  
C Fibers ◽  
Arterial Blood ◽  
C Fiber ◽  
Eosinophil Granule

Experiments were performed to test the hypothesis that human eosinophil granule-derived cationic proteins stimulate vagal C-fiber afferents in the lungs and elicit pulmonary chemoreflex responses in anesthetized Sprague-Dawley rats. Intratracheal instillation of eosinophil cationic protein (ECP; 1–2 mg/ml, 0.1 ml) consistently induced an irregular breathing pattern, characterized by tachypnea (change in breathing frequency of 44.7%) and small unstable tidal volume (Vt). The tachypnea, accompanied by decreased heart rate and arterial blood pressure, started within 30 s after the delivery of ECP and lasted for >30 min. These ECP-induced cardiorespiratory responses were completely prevented by perineural capsaicin treatment of both cervical vagi, which selectively blocked C-fiber conduction, suggesting the involvement of these afferents. Indeed, direct recording of single-unit activities of pulmonary C-fibers further demonstrated that the same dose of ECP evoked a pronounced and sustained (>30-min) stimulatory effect on pulmonary C-fibers. Furthermore, the sensitivity of these afferents to lung inflation was also markedly elevated after the ECP instillation, whereas the vehicle of ECP administered in the same manner had no effect. Other types of eosinophil granule cationic proteins, such as major basic protein and eosinophil peroxidase, induced very similar respiratory and cardiovascular reflex responses. In conclusion, these results show that eosinophil granule-derived cationic proteins induce a distinct stimulatory effect on vagal pulmonary C-fiber endings, which may play an important role in the airway hyperresponsiveness associated with eosinophil infiltration in the airways.

A light scattering study of interactions of oppositely charged proteins in solution

2007 ◽  
Vol 1061 ◽  
Author(s):  
Perumal Radha Ramasamy ◽  
Raafat Elmaghrabi ◽  
Gary Halada ◽  
Miriam Rafailovich
Keyword(s):  
Light Scattering ◽  
Zeta Potential ◽  
Fluorescent Dyes ◽  
Cationic Protein ◽  
Cationic Proteins ◽  
Low Concentrations ◽  
Scattering Study ◽  
Fluorescent Tags ◽  
Oppositely Charged ◽  
Electrophoresis Of Proteins

ABSTRACTIn experiments involving electrophoresis of proteins in gels, it was observed that the mobility of FITC tagged albumin (FITC albumin) was greater than that of TRITC tagged albumin (TRITC albumin). To further understand the effects of tagging proteins with fluorescent dyes, interactions of anionic proteins FITC albumin and untagged bovine serum albumin (BSA), with cationic protein ploy-L-lysine was studied using dynamical light scattering. It was found that aggregates formed by the interaction of FITC albumin with poly-L-lysine were larger than those formed by the interaction between poly-L-lysine and BSA. Using zeta potential measurements it was observed that irrespective of the fluorescent tags attached to them, the zeta potential values of cationic proteins changed from negative to positive with increasing amounts of poly-L-lysine. It was also observed that addition of small amounts of poly-L-lysine to solutions containing FITC albumin decreased the zeta potential drastically. To explain this data, we are proposing a model that suggests that low concentrations of poly-L-lysine serve as scaffold - like structures on which several FITC albumin molecules anchor. We conclude that FITC appears to change the surface charge of albumin significantly and thereby influencing its behavior in solution and its interaction with cationic poly-L-lysine.

scholarly journals Cationic Albumin Encapsulated DNA Origami for Enhanced Cellular Transfection and Stability

Materials ◽  
2019 ◽  
Vol 12 (6) ◽  
pp. 949 ◽  
Author(s):  
Xuemei Xu ◽  
Shiqi Fang ◽  
Yuan Zhuang ◽  
Shanshan Wu ◽  
Qingling Pan ◽  
...  
Keyword(s):  
Mammalian Cells ◽  
Biomedical Applications ◽  
Transfection Efficiency ◽  
Dna Origami ◽  
Biological Applications ◽  
Dna Nanostructures ◽  
Cationic Protein ◽  
Physiological Conditions ◽  
Cellular Transfection ◽  
Biological Environment

DNA nanostructures, owing to their controllable and adaptable nature, have been considered as highly attractive nanoplatforms for biomedical applications in recent years. However, their use in the biological environment has been restricted by low cellular transfection efficiency in mammalian cells, weak stability under physiological conditions, and endonuclease degradation. Herein, we demonstrate an effective approach to facilitate fast transfection of DNA nanostructures and enhance their stability by encapsulating DNA origami with a biocompatible cationic protein (cHSA) via electrostatic interaction. The coated DNA origami is found to be stable under physiological conditions. Moreover, the cHSA coating could significantly improve the cellular transfection efficiency of DNA origami, which is essential for biological applications.

scholarly journals PRODUCTION OF INFLAMMATORY CHANGES IN THE MICROCIRCULATION BY CATIONIC PROTEINS EXTRACTED FROM LYSOSOMES

1964 ◽  
Vol 120 (5) ◽  
pp. 747-764 ◽  
Author(s):  
Aaron Janoff ◽  
Benjamin W. Zweifach
Keyword(s):  
Protein Fraction ◽  
Basic Protein ◽  
Tissue Injury ◽  
Active Material ◽  
Supernatant Fraction ◽  
Absorption Data ◽  
Cationic Protein ◽  
Cationic Proteins ◽  
Inflammatory Changes ◽  
Ethanol Fraction

Lysosomal granules of rabbit exudate polymorphonuclear (PMN) leucocytes were isolated and then lysed by freezing-thawing. Topical application of this material to rat and rabbit mesentery produced sticking and emigration of leucocytes, stasis of blood flow, and petechial hemorrhage. The granule-free, supernatant fraction of the homogenized leucocytes failed to produce any of these reactions. Cationic proteins extracted from these granules by weak acid and precipitated by ethanol at concentrations of 20 and 45 per cent, were also tested on heterologous, homologous, and autologous mesenteric vessels. The 20 per cent ethanol-precipitated fraction produced all of the aforementioned injury reactions, whereas the 45 per cent fraction was inactive. The intensity of inflammatory changes produced by the active cationic protein fraction was greater than that produced by lysed whole granules. Both the 20 per cent and 45 per cent ethanol fractions of cationic protein induced clumping of rabbit platelets, in vitro. The 20 per cent ethanol fraction also caused a slight acceleration in rate of swelling of isolated rabbit liver mitochondria. The active material proved to be non-pyrogenic in rabbits. This material exhibited no kinin-like effects when tested on isolated smooth muscle preparations (rabbit aorta and guinea pig ileum). In the rat, the protein produced a transient vasodepression which was inhibited by pretreatment of the animal with an antihistamine. Ultraviolet absorption data and ribose assays showed that the 20 per cent ethanol fraction contained only 4 per cent or less of ribonucleic acid. Upon electrophoresis in starch gel, using acid buffer, this fraction separated into at least three major components which migrated towards the cathode. Precipitation of one of the slowly migrating components by titration of the fraction to pH 10.5 greatly increased the inflammatory activity of the material. The inflammatory basic protein fraction was essentially devoid of acid phosphatase, beta glucuronidase, acid ribonuclease, lysozyme, and catalase activity. The non-inflammatory basic protein fraction contained appreciable quantities of acid ribonuclease and lysozyme. The foregoing data demonstrate that certain of the cationic proteins present in lysosomes of rabbit exudate PMN leucocytes can reproduce one of the cardinal features of the inflammatory response; namely, adhesion and emigration of leucocytes in the microcirculation. These findings offer fresh support for the role of lysosomes in the pathogenesis of tissue injury, and may help to account for the propagation of leucocyte emigration to peak numbers during inflammatory reactions.

Cationic neutrophil proteins increase transendothelial albumin movement

1987 ◽  
Vol 62 (4) ◽  
pp. 1521-1530 ◽  
Author(s):  
M. W. Peterson ◽  
P. Stone ◽  
D. M. Shasby
Keyword(s):  
Human Neutrophil ◽  
Cell Shape ◽  
Human Neutrophil Elastase ◽  
Gap Formation ◽  
Cationic Protein ◽  
Edema Formation ◽  
Cationic Proteins ◽  
Fetal Bovine ◽  
Secretory Products ◽  
Shape Changes

Neutrophils play a role in the development of pulmonary edema in many models of the adult respiratory distress syndrome, but the mechanism of their action is not completely understood. We asked whether two neutrophil secretory products, human neutrophil cationic protein (NCP) and human neutrophil elastase (HNE), would nonenzymatically alter the movement of albumin across a cultured endothelial monolayer. Both enzymes were inactivated by heating before use. HNE was additionally enzymatically inactivated with a chloromethylketone oligopeptide (CMK) inhibitor and with alpha 1-proteinase inhibitor (alpha 1-PI). Heated NCP, heated HNE, and CMK-complexed HNE all increased transendothelial albumin transfer. The cation protamine also increased albumin transfer across the endothelium and this increase was blocked by heparin. Alpha 1-PI and fetal bovine serum also prevented the cationic proteins from increasing albumin transfer. Using the release of lactate dehydrogenase as a marker of cytotoxicity, heated HNE was toxic to endothelial cells, heated NCP had only minimal toxicity, and protamine had no toxicity. Changes in endothelial cell shape with gap formation was seen after exposure to both heated HNE and heated NCP. Both the cytotoxicity associated with heated HNE and the cell shape changes associated with heated NCP and heated HNE could be blocked by heparin. These results suggest that in addition to neutrophil proteases and reactive O2 molecules, neutrophil-derived cationic proteins can directly and nonenzymatically contribute to edema formation during acute inflammation.

scholarly journals Impact of dichloroethane on the microbicidal activity of neutrophils and mononuclear phagocytes

2017 ◽  
Vol 98 (3) ◽  
pp. 415-418
Author(s):  
I A Men’shikova ◽  
N A Mufazalova ◽  
F Kh Kamilov ◽  
L F Mufazalova
Keyword(s):  
Total Dose ◽  
Lethal Dose ◽  
Peritoneal Macrophages ◽  
Mononuclear Phagocytes ◽  
Myeloperoxidase Activity ◽  
Cationic Protein ◽  
Nonspecific Resistance ◽  
Cationic Proteins ◽  
Microbicidal Activity ◽  
Colony Forming Units

Aim. To study the damaging effect of dichloroethane on the functional state of neutrophils and peritoneal macrophages. Methods. Dichloroethane was administered to animals intragastrically daily in olive oil at a dose of 0.84 mg/kg of body weight for 60 days so that the total dose was 0.1 of 50% lethal dose. The number of leukocytes, neutrophils and lymphocytes in peripheral blood, intensity of oxygen-dependant metabolism (induced test with nitro blue tetrazolium), antimicrobial activity in the conditions of functioning and blockade (by sodium azide), oxygen-dependent factors of microbicidity, content of myeloperoxidase and cationic proteins in neutrophils and peritoneal macrophages were measured. The results were recorded the next day after introduction of the toxicant. Fungicidal activity was measured by the number of colony-forming units of C. albicans, growing on day 3 at culture medium. Results. Intoxication with dichloroethane for 60 days leads to the formation of leukopenia, mainly due to the decreased number of neutrophils while reducing the number of lymphocytes. This is accompanied by inhibition of oxygen-dependant killing of neutrophils as a result of suppression of peroxidase-dependant mechanisms of microbicidity (the formation of oxygen active forms decreases). Besides, decrease of activity of oxygen-independant mechanisms of killing develops, which correlates with a reduction of cationic proteins level. Suppression of oxidative and non-oxidative mechanisms of microbicidity of peritoneal macrophages. This is accompanied by a decrease of oxygen-dependant metabolism intensity, myeloperoxidase activity and cationic protein level in these cells. Conclusion. Intoxication with dichloroethane for 60 days in a total dose of 0.1 of 50% lethal dose has a profound damaging effect on the cells of phagocytic link of nonspecific resistance: formation of leukopenia, suppression of oxidative metabolism and microbicidal activity of neutrophils and mononuclear phagocytes occur.

Applications of Scanning Microscopy in Biomedical Research

1968 ◽  
Vol 26 ◽  
pp. 354-355
Author(s):  
T. L. Hayes
Keyword(s):  
Information Content ◽  
Biomedical Research ◽  
Biomedical Applications ◽  
Three Dimensional ◽  
Dental Enamel ◽  
Resolving Power ◽  
Whole Mount ◽  
Two Parameters ◽  
Content Information ◽  
Sectioned Tissue

Biomedical applications of the scanning electron microscope (SEM) have increased in number quite rapidly over the last several years. Studies have been made of cells, whole mount tissue, sectioned tissue, particles, human chromosomes, microorganisms, dental enamel and skeletal material. Many of the advantages of using this instrument for such investigations come from its ability to produce images that are high in information content. Information about the chemical make-up of the specimen, its electrical properties and its three dimensional architecture all may be represented in such images. Since the biological system is distinctive in its chemistry and often spatially scaled to the resolving power of the SEM, these images are particularly useful in biomedical research.In any form of microscopy there are two parameters that together determine the usefulness of the image. One parameter is the size of the volume being studied or resolving power of the instrument and the other is the amount of information about this volume that is displayed in the image. Both parameters are important in describing the performance of a microscope. The light microscope image, for example, is rich in information content (chemical, spatial, living specimen, etc.) but is very limited in resolving power.

How SIMS microscopy can be used in medicine

1992 ◽  
Vol 50 (2) ◽  
pp. 1602-1603
Author(s):  
Philippe Fragu
Keyword(s):  
Mass Spectrometry ◽  
Biomedical Applications ◽  
Secondary Ion Mass Spectrometry ◽  
Chemical Elements ◽  
Biological Applications ◽  
The Past ◽  
Potential Source ◽  
Secondary Ion ◽  
Chemical Integrity ◽  
Scientific Endeavour

The identification, localization and quantification of intracellular chemical elements is an area of scientific endeavour which has not ceased to develop over the past 30 years. Secondary Ion Mass Spectrometry (SIMS) microscopy is widely used for elemental localization problems in geochemistry, metallurgy and electronics. Although the first commercial instruments were available in 1968, biological applications have been gradual as investigators have systematically examined the potential source of artefacts inherent in the method and sought to develop strategies for the analysis of soft biological material with a lateral resolution equivalent to that of the light microscope. In 1992, the prospects offered by this technique are even more encouraging as prototypes of new ion probes appear capable of achieving the ultimate goal, namely the quantitative analysis of micron and submicron regions. The purpose of this review is to underline the requirements for biomedical applications of SIMS microscopy.Sample preparation methodology should preserve both the structural and the chemical integrity of the tissue.

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