STUDY OF ACTIVATING EFFECT OF SURFACTANT DRUGS ON PULMONARY MACROPHAGES IN VITRO IN PATIENTS WITH BRONCHIECTASIS

2020 ◽  
pp. 5-8
Author(s):  
Dmitrii Valerevich Minukhin
Keyword(s):  
Cell Surface ◽  
Bronchoalveolar Lavage ◽  
Injury Rate ◽  
Exogenous Surfactant ◽  
Control Group ◽  
Electron Microscopic ◽  
Local Immunity ◽  
Surfactant System ◽  
Pulmonary Macrophages

The state of surfactant system of lungs in chronic inflammation is known to disrupt their structural organization, causes shifts in the system itself, the severity of which depends on alveolar parenchyma injury rate. The cell destruction of aerohematical barrier is accompanied by changes in pulmonary surfactant system, its deficiency, development of dis− and atelectases, which increases the severity of lung injury and promotes the progression of bronchiectasis. Exogenously introduced surfactant restores local immunity and serves as a substrate for synthesis of its own surfactant. To determine the stimulatory effect of natural surfactant on receptors of the lung macrophages and its influence on activation of phagocytosis a comparative study of direct effect of infasurf on cell surface and formation of phagocytic vacuoles in macrophage elements of individuals with bronchodectic disease at an acute stage was performed . In bronchoalveolar lavage of patients the percentage and viability of pulmonary macrophages were determined. Other lavage material was investigated both as a baseline control and with infasurf. The electron microscopic examination of bronchoalveolar lavage macrophage elements in patients show that after incubation of cells with infasurf, there is a pronounced cell surface activation, formation of phagocytic vacuoles containing membranes of exogenous surfactant. Its small and large clusters were often observed lying freely between pulmonary macrophages, whereas in the control group (without infasurf) small fragments of tubular myelin or osmiophilous lamellar bodies were noted. Thus, the specific pharmacological action of surfactant drugs directly contributes to differentiation and maturation of pulmonary macrophages, their phagocytic function activation. Key words: bronchiectasis, surfactant, pulmonary macrophages.

An integrated screening method for evaluating the pulmonary toxicity of inhaled particulates: Role of microscopic techniques in assessing pulmonary macrophage clearance functions

1990 ◽  
Vol 48 (3) ◽  
pp. 826-827
Author(s):  
David B. Warheit ◽  
Lena Achinko ◽  
Mark A. Hartsky
Keyword(s):  
Bronchoalveolar Lavage ◽  
Pulmonary Toxicity ◽  
Screening Method ◽  
Pulmonary Response ◽  
Inhaled Particles ◽  
Cytochemical Analysis ◽  
Pulmonary Macrophages ◽  
Integrated Screening

There is a great need for the development of a rapid and reliable bioassay to evaluate the pulmonary toxicity of inhaled particles. A number of methods have been proposed, including lung clearance studies, bronchoalveolar lavage analysis, and in vitro cytotoxicity tests. These methods are often limited in scope inasmuch as they measure only one dimension of the pulmonary response to inhaled, instilled or incubated dusts. Accordingly, a comprehensive approach to lung toxicity studies has been developed.To validate the method, rats were exposed for 6 hours or 3 days to various concentrations of either aerosolized alpha quartz silica (Si) or carbonyl iron (CI) particles. Cells and fluids from groups of sham and dust-exposed animals were recovered by bronchoalveolar lavage (BAL). Alkaline phosphatase, LDH and protein values were measured in BAL fluids at several time points postexposure. Cells were counted and evaluated for viability, as well as differential and cytochemical analysis. In addition, pulmonary macrophages (PM) were cultured and studied for morphology, chemotaxis, and phagocytosis by scanning electron microscopy.

Lipocortin-1 distribution in bronchoalveolar lavage from healthy human lung: effect of prednisolone

1995 ◽  
Vol 79 (1) ◽  
pp. 121-128 ◽  
Author(s):  
S. F. Smith ◽  
T. D. Tetley ◽  
A. K. Datta ◽  
T. Smith ◽  
A. Guz ◽  
...  
Keyword(s):  
Cell Surface ◽  
Bronchoalveolar Lavage ◽  
Ex Vivo ◽  
Specific Cell ◽  
Surface Binding ◽  
Healthy Human ◽  
Before And After ◽  
The Difference ◽  
Specific Cell Surface

Lipocortin-1 (LC-1; annexin-1) may mediate some anti-inflammatory actions of the glucocorticoids, probably after binding to specific cell surface binding sites. We have quantified LC-1 levels in bronchoalveolar lavage (BAL) fluid and cells collected from seven healthy volunteers before and after 7 days of treatment with an oral glucocorticoid, prednisolone (30 mg/day). Extracellular BAL LC-1 was higher and cellular LC-1 was lower after prednisolone than before [extracellular: before, median 98 ng/mg albumin (range 48–350 ng/mg albumin); after, 236 ng/mg albumin (19–414 ng/mg albumin); P < 0.05. Cellular: before, 23.3 ng/10(6) cells (14.6–26.9 ng/10(6) cells); after, 18.0 ng/10(6) cells (122–268 ng/10(6) cells); P < 0.05]. The distribution of LC-1 within BAL cells ex vivo (cell surface = 25%, cytosol = 50%, membrane = 25%) was unaffected by prednisolone treatment. However, in adherent cells that had been cultured for 4 h, 70–80% of the LC-1 was on the cell surface. In summary, prednisolone appears to promote cellular release of LC-1. The difference in distribution of cellular LC-1 in BAL cells ex vivo and in vitro may reflect adherence and/or activation.

scholarly journals An In Vitro Microbial-Caries Model Used to Study the Efficacy of Antibodies to Streptococcus mutans Surface Proteins in Preventing Dental Caries

2000 ◽  
Vol 7 (1) ◽  
pp. 49-54 ◽  
Author(s):  
Margherita Fontana ◽  
Tiffany L. Buller ◽  
Ann J. Dunipace ◽  
George K. Stookey ◽  
Richard L. Gregory
Keyword(s):  
Cell Surface ◽  
Streptococcus Mutans ◽  
Positive Control ◽  
Surface Proteins ◽  
Laser Scanning Microscopy ◽  
Control Group ◽  
Lesion Area ◽  
Lesion Depth ◽  
Caries Model

ABSTRACT The first step for a pathogenic bacterium to initiate infection is via attachment (i.e., through surface determinants) to a suitable receptor. An in vitro microbial artificial-mouth model was used to test the efficacy of polyclonal antibodies to Streptococcus mutans cell surface proteins (CsAb) and a cell surface 59-kDa protein (59Ab) in preventing S. mutans colonization and carious lesion formation. In study 1, groups of 12 human teeth specimens were inoculated with S. mutans, which were incubated with different concentrations of CsAb (A1 [positive control], sterile saline, no antibody; A2, 0.007 mg of antibody protein/ml; and A3, 0.7 mg of antibody protein/ml) for 1 h at 37°C. The negative control group (B1) was not infected and was incubated with Trypticase soy broth (TSB) without dextrose supplemented with 5% sucrose (TSBS). In study 2, the same study design was used except that 59Ab was used instead of CsAb, normal rabbit serum was used in the positive control group (A1), and TSB supplemented with 1% glucose was used as the nutrient to control for sucrose-dependent colonization. All groups were exposed for 4 days to circulating cycles of TSBS and TSB (study 1 and study 2, respectively; 30 min each, three times per day) and a mineral washing solution (21 h per day). Prior to each nutrient cycle, 1 ml of the appropriate CsAb or 59Ab solution was administered to each group and allowed to mix for 30 min before cycling was resumed. Data obtained by confocal laser scanning microscopy demonstrated the presence of a significantly smaller (P < 0.05) lesion area and a smaller total lesion fluorescence in group A3 than in group A1 for both studies. In study 1, group A2 had significantly smaller values than A1 for lesion depth and area. There were no significant differences between groups A2 and A3 for lesion area or between groups A1 and A2 for total lesion fluorescence. In study 2, there were no significant differences among groups A1 and A2 for lesion depth or between groups A2 and A3 for all of the parameters studied. In both studies, there were no significant differences between S. mutans plaque CFU numbers among any of the groups. These studies demonstrated the efficacy of CsAb and 59Ab in reducing primary caries development in this model, although the underlying mechanism remains unclear.

scholarly journals Microhardness and Scanning Electron Microscopic Morphology of Permanent Dentin Following the Application of Iranian and Foreign-Made Desensitizing Toothpastes

2020 ◽  
Vol 12 (3) ◽  
pp. 86-92
Author(s):  
Azam Valian ◽  
Mina Jafari ◽  
Hosna Ebrahimizadeh
Keyword(s):  
Repeated Measures ◽  
Negative Control ◽  
Superior Performance ◽  
Control Group ◽  
Electron Microscopic ◽  
Significant Difference ◽  
The Mean ◽  
Microscopic Morphology ◽  
Scanning Electron

Background: This study aimed to assess the microhardness and morphology of permanent dentin following the application of Iranian and foreign-made desensitizing toothpastes. Methods: This in vitro, experimental study evaluated 48 dentin samples prepared of the extracted sound human permanent molars. Dentin samples were randomly divided into three groups (n=16) and subjected to the application of Pooneh Iranian desensitizing toothpaste, Colgate Sensitive Pro- Relief (Poland), and no intervention (negative control). Each dentin sample was then immersed in 5 mL of a demineralizing solution for 10 hours and underwent a demineralization/remineralization cycle (pH cycling) for 14 days. The mean microhardness of the samples was measured at baseline after demineralization and after 14 days. One sample of each toothpaste group was selected for the scanning electron microscopy (SEM) assessment of dentin morphology. Finally, repeated-measures ANOVA was used to analyze the effect of time and type of toothpaste on microhardness using SPSS, version 21. Results: The comparison of the mean microhardness of the control group with experimental groups revealed no significant difference at baseline or after demineralization (P > 0.05) although this difference was significant after 14 days (P < 0.001). The mean microhardness of the two toothpaste groups was not significantly different at different time points (P > 0.05). Finally, the SEM assessment revealed a greater tubular obstruction in Pooneh group after 14 days. Conclusions: Pooneh Iranian-made desensitizing toothpaste was comparable to Colgate Sensitive Pro-Relief foreign-made desensitizing toothpaste in terms of the microhardness of permanent dentin. It even demonstrated superior performance with regard to the obstruction of dentinal tubules.

In vitro genotoxicity assessment and comparison of cerium (IV) oxide micro- and nanoparticles

2020 ◽  
Vol 36 (2) ◽  
pp. 76-83 ◽  
Author(s):  
Kader Arslan ◽  
Giray Buğra Akbaba
Keyword(s):  
Human Peripheral Blood ◽  
Microscopic Analysis ◽  
Negative Control Group ◽  
Negative Control ◽  
Control Group ◽  
Average Particle ◽  
Particle Sizes ◽  
Electron Microscopic ◽  
Scanning Electron Microscopic Analysis

Cerium (IV) oxide (CeO2), which is used as a biomaterial, has wide application in areas such as the biomedical, glass polishing, electronic, automotive, and pharmacology industries. Comparing with the literature, in this study, the genotoxic effects of cerium (IV) oxide microparticles (COMPs) and cerium (IV) oxide nanoparticles (CONPs) were investigated for the first time in human peripheral blood cultures at concentrations of 0.78, 1.56, 3.125, 6.25, 12.5, 25, and 50 ppm for 72 h under in vitro conditions. Particle sizes of COMPs and CONPs were determined using scanning electron microscopic analysis. Micronucleus and chromosome aberration tests were used to determine the genotoxicity of COMPs and CONPs. The average particle sizes of COMPs and CONPs were approximately 148.25 and 25.30 nm, respectively. It was determined that CeO2 particles in both micro and nano sizes were toxic at all concentrations compared to the negative control group (distilled water). Importantly, COMPs and CONPs were genotoxic even at the lowest concentration (0.78 ppm). Comparing particle sizes, the data indicated that COMPs were more toxic than CONPs. The results suggest that genotoxicity of COMPs and CONPs may be a function of applied concentrations and particle sizes.

ASSESSMENT OF THE LUNG NEUTROPHILS AND MACROPHAGES FUNCTIONAL STATE IN COMMUNITY-ACQUIRED PNEUMONIA

2021 ◽  
pp. 23-29
Author(s):  
Kislova S.E. ◽  
Kondrashova N.M. ◽  
Plekhova N.G.
Keyword(s):  
Functional Activity ◽  
Sharp Decrease ◽  
Community Acquired Pneumonia ◽  
Absorption Capacity ◽  
Induced Sputum ◽  
Phagocytic Index ◽  
Control Group ◽  
Local Immunity ◽  
Mean Values

The progression of the inflammatory process and the outcome in community-acquired pneumonia (CAP) depend on the functional activity of innate immune cells - neutrophils and macrophages. Purpose: to establish the relationship between the morphofunctional state of neutrophils and macrophages of induced sputum (inflammation focus) and the severity of community-acquired pneumonia (CAP). Materials and methods: cells were removed from the induced sputum (IS) of healthy individuals and patients with CAP. In the «in vitro» model, upon contact with S. aureus the phagocytic index (PI), number (PN), and index of phagocytosis completion (ICP) were determined. A spectrophotometric method was used to assess the enzymatic activity of cells. Results. The dependence of phagocytosis indices on the severity of CAP was established (p <0.05). A sharp decrease in the phagocytic and functional activity of cells was recorded in the focus of inflammation in patients with severe pneumonia. It was found that in patients with CAP, compared with the control group, a significant decrease in the relative content of cells capable of reducing nitro-blue tetrazolium was found. In all groups of patients with this disease, the reserve capacity for myeloperoxidase (MPO) had negative values, which proved a decrease in the reactivity of local immunity cells. The MPO values for the cells of patients with a moderate course of the disease were higher than in the rest of the examined individuals, which indicated the activation of the bactericidal activity of neutrophils in MI. In patients with severe CAP in the spontaneous test, the mean values of MPO activity were 2.2 times higher than in the control group. Conclusion: a comprehensive study of innate immunity cells functional activity in induced sputum, including the quantitative determination of phagocytic parameters with an assessment of the absorption capacity and completeness of phagocytosis, makes it possible to assess the severity of community-acquired pneumonia.

Effect of Various Concentrations of Sodium Hypochlorite on Primary Dentin: An in vitro Scanning Electron Microscopic Study

2012 ◽  
Vol 37 (1) ◽  
pp. 37-43 ◽  
Author(s):  
L Gowda ◽  
U Mohan Das
Keyword(s):  
Sodium Hypochlorite ◽  
In Vitro Study ◽  
Complete Removal ◽  
Collagen Fibrils ◽  
Smear Layer ◽  
Control Group ◽  
Electron Microscopic ◽  
Time Of Application ◽  
Anterior Teeth

Background: Sodium hypochlorite solutions have been evaluated for their effects in bonding procedures as they are found to deplete or remove the organic portion of the dentin, particularly the collagen fibrils. The aim of this in vitro study was to assess and compare the efficacies of 1%, 2.5%, 5% and 10% NaOCl at 30, 60 and 120s on etched primary dentin. Methods: 84 primary anterior teeth were ground to expose a flat dentin area on the buccal surface. The specimens were divided into fourteen groups of six each based on the dentin surface treatment (35% phosphoric acid etching for 7 seconds-AE and/or NaOCl application), NaOCl solution concentrations (1%, 2.5%, 5% and 10%) and time of application (0, 30, 60 and 120s). Specimens were prepared for SEM and photomicrographs were taken of the surface and were scored against a five point scale, based on the smear layer and amount of collagen removed. The scores were submitted to Kruskal-Wallis and Mann Whitney tests. Results: This study showed the presence of smear layer in the control group. The group treated with Acid Etchant showed a demineralized pattern of dentin with exposure of dentin tubules and collagen fibrils network on the intertubular and peritubular dentin which was not significantly different from the groups treated with 1% and 2.5% NaOCl. Groups treated with 5% NaOCl were not statistically different from each other, the surface was corroded but collagen fibrils were not completely depleted. Groups treated with 10% NaOCl were not statistically different from each other and showed complete removal of collagen fibrils with wider tubular apertures and several secondary tubules on peritubular and intertubular dentin. Conclusion: Higher concentrations of NaOCl solutions (5% and 10%) produced significant changes in the etched primary dentin. The higher the concentration of the NaOCl solution, the lower can be the time for the application of the solution for the complete removal of collagen fibrils.

scholarly journals Cytotoxicity and Oxidative Stress Responses of Imidacloprid and Glyphosate in Human Prostate Epithelial WPM-Y.1 Cell Line

2020 ◽  
Vol 2020 ◽  
pp. 1-12
Author(s):  
Khaled Y. Abdel-Halim ◽  
Safaa R. Osman
Keyword(s):  
Oxidative Stress ◽  
Cell Line ◽  
Stress Responses ◽  
Agricultural Sector ◽  
In Vitro Assays ◽  
Control Group ◽  
Electron Microscopic ◽  
Ic50 Values ◽  
And Oxidative Stress

Insecticide imidacloprid and herbicide glyphosate have a broad spectrum of applicable use in the agricultural sector of Egypt. Their ability to induce in vitro cytotoxic and oxidative stress on normal human cells (prostate epithelial WPM-Y.1 cell line) was evaluated with the methyl tetrazolium test (MTT) and histopathological investigation. Cell viability was evaluated with an MTT test for 24 h. The median inhibition concentration (IC50) values were 0.023 and 0.025 mM for imidacloprid and glyphosate, respectively. Sublethal concentrations: 1/10 and 1/50 of IC50 and IC50 levels significantly induced an increase in the lactate dehydrogenase (LDH) activity and malondialdehyde (MDA) level compared with the untreated cells. Rapid decrease in the glutathione (GSH) content and glutathione-S-transferase (GST) activity was induced. Significant increases were recorded in activities of catalase (CAT), glutathione peroxidase (GPx), and glutathione reductase (GR), respectively, compared with the control group. Transmission electron microscopic (TEM) investigation showed significant defects in the cells following pesticide treatments for 24 h. Therefore, it is concluded that imidacloprid and glyphosate are very toxic in vitro assays and able to induce apoptotic effects as well as oxidative stress. So, these findings provide a scenario of multibiomarkers to achieve the imposed risks of pesticides at low doses.

Inhaled nitric oxide injures the pulmonary surfactant system of lambs in vivo

1996 ◽  
Vol 270 (2) ◽  
pp. L273-L280 ◽  
Author(s):  
S. Matalon ◽  
V. DeMarco ◽  
I. Y. Haddad ◽  
C. Myles ◽  
J. W. Skimming ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Bronchoalveolar Lavage ◽  
Surface Properties ◽  
Pulmonary Surfactant ◽  
Protein A ◽  
Surfactant Protein ◽  
Surfactant System ◽  
Minimum Surface Tension

Nitric oxide (.NO) is a free radical, and as such may damage the pulmonary surfactant system. To determine the potential toxicity of .NO in vivo, we exposed 35 newborn lambs to 0, 20, 80 or 200 ppm .NO in either 21 or 60% O2 for 6 h. At the end of the exposure, lambs had normal values of arterial Po2, Pco2, and pH; total protein concentration in the bronchoalveolar lavage was also at normal levels. There were no differences in the surface properties of surfactant among the air or 60% O2 groups. Pulmonary surfactant samples, isolated from the bronchoalveolar lavage of lambs breathing air or 20 ppm .NO and reconstituted at a lipid concentration of 3 mg/ml, reached a low minimum surface tension (Tmin < 3 mN/m) in a pulsating bubble surfactometer. On the other hand, abnormal surface properties were observed in 36 and 60% of surfactant samples isolated from lungs of lambs that breathed 80 or 200 ppm .NO, respectively. These findings were confirmed using a captive bubble surfactometer. Surfactant protein A, isolated from the lungs of lambs that breathed 200 ppm .NO, exhibited decreased ability to aggregate lipids in vitro. These data are consistent with injury to the surfactant apoproteins during inhalation of either 80 or 200 ppm .NO for 6 h.

Cytoplasmic Granules in Lymphocytes from Rats Dosed with SK&F 14336-D

1971 ◽  
Vol 29 ◽  
pp. 556-557
Author(s):  
T. G. Merrill ◽  
B. J. Payne ◽  
A. J. Tousimis
Keyword(s):  
Lipid Storage ◽  
Pokeweed Mitogen ◽  
Electron Microscopic ◽  
Cytoplasmic Granules ◽  
Storage Diseases ◽  
Tay Sachs Disease ◽  
Large Numbers ◽  
Microscopic Studies ◽  
Neurovisceral Lipidosis

Rats given SK&F 14336-D (9-[3-Dimethylamino propyl]-2-chloroacridane), a tranquilizing drug, developed an increased number of vacuolated lymphocytes as observed by light microscopy. Vacuoles in peripheral blood of rats and humans apparently are rare and are not usually reported in differential counts. Transforming agents such as phytohemagglutinin and pokeweed mitogen induce similar vacuoles in in vitro cultures of lymphocytes. These vacuoles have also been reported in some of the lipid-storage diseases of humans such as amaurotic familial idiocy, familial neurovisceral lipidosis, lipomucopolysaccharidosis and sphingomyelinosis. Electron microscopic studies of Tay-Sachs' disease and of chloroquine treated swine have demonstrated large numbers of “membranous cytoplasmic granules” in the cytoplasm of neurons, in addition to lymphocytes. The present study was undertaken with the purpose of characterizing the membranous inclusions and developing an experimental animal model which may be used for the study of lipid storage diseases.

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