scholarly journals Galectin-8 Modulates Innate and Adaptive Immune Response Genes in Bovine Neutrophils

2019 ◽  
Vol 9 (1) ◽  
pp. 24 ◽  
Author(s):  
Eboghoye Eluka-Okoludoh ◽  
Kingsley Ekwemalor ◽  
Sarah Adjei-Fremah ◽  
Bharath Mulakala ◽  
Mulumebet Worku
Keyword(s):  
Immune Response ◽  
Total Protein ◽  
Protein Concentration ◽  
Neutrophil Migration ◽  
Transcript Abundance ◽  
Fold Change ◽  
Total Protein Concentration ◽  
Adaptive Immune ◽  
Immune Response Genes ◽  
Bovine Neutrophils

Galectins (Gals) are a family of animal lectins that bind β-galactosides through a carbohydrate recognition domain. Galectin-8 is a tandem-repeat galectin, secreted intracellularly and extracellularly. It is associated with neutrophil migration and has been studied as a possible therapeutic to combat inflammation. The objective of this study was to evaluate the translational and the transcriptional effects of recombinant Galectin-8 (rGal-8) on cow neutrophils. Blood was collected aseptically from Holstein-Friesian cows (n=10) from the North Carolina A&T State University Dairy Unit. Neutrophils isolated were treated with rGal-8 (2μg), or PBS (control) and were incubated at 37°C, 5% CO2 for 1 hour. Supernatant from treated neutrophils was evaluated for total protein concentration, and galectin-8 secretion using bovine Galectin-8 Enzyme Linked-Immuno-Sorbent Assay (ELISA) kit. Total RNA was extracted, reverse transcribed, and RT-qPCR was performed using the RT² Profiler Cow Innate & Adaptive Immune Responses Array with 84 genes. The Livak method was used to calculate transcript abundance and fold change (FC>2 considered significant). Total protein concentration increased (P=0.0361) after rGal-8 treatment compared to the untreated control. Galectin-8 secretion was not significantly different in control compared to treated group (P=0.5819). Out of the 84 genes, 81 genes were differentially expressed in response to rGal-8; 14 up-regulated, 5 down-regulated, 61 genes remained unchanged. Treatment with rGal8 induced the expression of IRF7. The top five up-regulated genes include FAS, CD40, CD86, IFNGR1, STAT1; down-regulated genes were TLR9, CD14, CCR6, TICAM1, and TLR1. Selected genes were probed to validate fold change; the levels of gene expression were comparable to data from RT2 array. Exposure of bovine neutrophils to rGal-8 modified expression of immune response genes. The functional significance of the change needs further studies.

scholarly journals Local and Systemic Up-regulation of TNFα, IL-1β and IL-6 in Mice Intratracheally Inoculated with Porphyromonas gingivalis

2008 ◽  
Vol 77 (3) ◽  
pp. 377-385 ◽  
Author(s):  
Z. Pavlica ◽  
A. Nemec ◽  
A. Nemec-Svete ◽  
D. Eržen ◽  
D. A. Crossley ◽  
...  
Keyword(s):  
Immune Response ◽  
Immune Responses ◽  
Porphyromonas Gingivalis ◽  
Total Protein ◽  
Protein Concentration ◽  
Serum Levels ◽  
Treated Group ◽  
Control Group ◽  
Total Protein Concentration ◽  
Sterile Phosphate Buffer

The objective of the study was to find whether a single intratracheal inoculation with live Porphyromonas gingivalis ATCC 33277 influences local and systemic inflammatory and immune responses in mice.Twelve-week-old BALB/c mice were intratracheally inoculated with 2.9 × 109 CFU P. gingivalis ATCC 33277 diluted in 40 μl sterile phosphate buffer (treated group) or with sterile PBS (control group). The animals were sacrificed 2, 6, 24, 72 and 168 h after inoculation. TNFα, IL-1β, IL-6 and total protein concentrations were measured in the serum, lungs and kidneys. Six hours after P. gingivalis inoculation, TNFα concentration was significantly increased in serum (p = 0.02) and kidneys (p = 0.04), but in the lungs TNFα production was enhanced already 2 h (p < 0.0001) after inoculation, reaching the peak after 6 h (p < 0.0001). The IL-1β concentration was also significantly increased in serum after 2 h (p = 0.006), remaining significantly elevated up to 3 days (p ≤ 0.0001) after inoculation. In lungs IL-1β levels were significantly increased 6 and 24 h (p < 0.0001) and in kidneys 24 h (p < 0.0001) and 168 h (p = 0.01) after inoculation. The IL-6 concentration was significantly increased in serum after 72 and 168 h (p < 0.0001). However, IL-6 was significantly increased in lungs after 6 h (p < 0.0001), remaining elevated until 72 h and in kidneys 2 and 6 h (p < 0.0001) after inoculation. Significantly increased total protein concentration was detected in kidneys 6 and 24 h (p < 0.0001) after inoculation. These results suggest that a single intratracheal inoculation with P. gingivalis stimulates the local and systemic inflammatory and immune response, as shown by increased tissue and serum levels of proinflammatory cytokines.

Determination of Total Protein Concentration in Solution Using Gold Electrode Modified with Silver Nanoparticles

2014 ◽  
Vol 27 (1) ◽  
pp. 253-257 ◽  
Author(s):  
Patrick Marcel Seumo Tchekwagep ◽  
Charles Péguy Nanseu-Njiki ◽  
Emmanuel Ngameni ◽  
Ravi Danielsson ◽  
Thomas Arnebrant ◽  
...  
Keyword(s):  
Silver Nanoparticles ◽  
Total Protein ◽  
Gold Electrode ◽  
Protein Concentration ◽  
Total Protein Concentration

Albumin fraction and measurement of total protein concentration

1993 ◽  
Vol 264 (5) ◽  
pp. H1723-H1726 ◽  
Author(s):  
B. T. Peterson ◽  
R. W. Tate
Keyword(s):  
Total Protein ◽  
Protein Concentration ◽  
Gamma Globulin ◽  
Colorimetric Assay ◽  
Full Range ◽  
Computational Method ◽  
Total Protein Concentration ◽  
Albumin Fraction ◽  
Standard Curve ◽  
Protein Assays

The standard curve of a typical colorimetric assay for total protein is often nonlinear and dependent on the albumin fraction of the protein standard. We developed a simple mathematical transformation to make the standard curve linear and a computational method to correct for differences in albumin concentrations among the samples. This method uses data from total protein assays on two sets of standards (albumin and gamma globulin) and provides accurate measures of total protein over the full range of albumin fractions. Comparison of this two-standard method with the a method that uses only albumin as a standard shows that this method prevents physiologically significant overestimations in total protein concentration and calculated protein osmotic pressure differences in the lungs.

scholarly journals Role of fibronectin under conditions of doxorubicin action

2015 ◽  
Vol 6 (1) ◽  
pp. 17-22
Author(s):  
A. I. Shevtsova ◽  
G. A. Ushakova
Keyword(s):  
Blood Plasma ◽  
Total Protein ◽  
Protein Concentration ◽  
Male Rats ◽  
Antitumor Action ◽  
Total Protein Concentration ◽  
Monospecific Antibodies ◽  
Anthracycline Chemotherapy ◽  
In Blood Plasma

There is no standard as to treatment of anthracycline chemotherapy complications. The reduction of cytotoxic drugs toxicity without weakening of their antitumor action remains relevant. The extracellular matrix which key component is fibronectin is present in all tissues and it continuously undergoes controlled remodeling. So, the purpose of our work was to study the level of fibronectin in the experimental model of doxorubicin-induced cardiomyopathy and effects of this cytostatic and its co-administration with antioxidants of different nature.The level of fibronectin was measured by ELISA using monospecific antibodies against fibronectin (Sigma, USA), secondary anti-IgG labeled with horseradish peroxidase (Sigma, USA) and fibronectin standard (Sigma, USA). The study was conducted on Wistar male rats with weight of 210 ± 50 g which were divided into 4 groups by 8 animals in each group: 1 – control, rats receiving saline i/p; 2 – doxorubicin 1 mg/kg i/p once a week during 4 weeks; 3 – doxorubicin by the same scheme plus 1% 2-oxoglutarate in drinking water during 4 weeks;4 – doxorubicin by the same scheme and korvitin injection 30 min before doxorubicin application once a week during 4 weeks. Obtained data indicate the effect of doxorubicin to decrease in index mass heart in 38% of animals compared to control animals; decrease in total protein concentration by 8% (Р < 0,05) and increase of the level of fibronectin by 67% (P < 0,001) in blood plasma of rats and decrease in the level of fibronectin in the heart extract by 19% (Р < 0,05) under development of doxorubicin-induced cardiotoxicity. Increased fibronectin concentration in blood plasma had strong correlation with decreased total protein concentration in blood (r=0,80) and heart extract (r=0,59) in rats with doxorubicin-induced cardiomiophaty indicating the sensitive reaction of fibronectin to development of metabolic disorders under doxorubicin influence. 

Assay of cerebrospinal fluid protein: a rate biuret method evaluated.

1983 ◽  
Vol 29 (1) ◽  
pp. 126-129 ◽  
Author(s):  
P R Finley ◽  
R J Williams
Keyword(s):  
Cerebrospinal Fluid ◽  
Total Protein ◽  
Protein Concentration ◽  
Protein A ◽  
Colorimetric Method ◽  
Total Protein Concentration ◽  
Reaction Cell ◽  
Biuret Method ◽  
Automated Instrument ◽  
Cerebrospinal Fluid Protein

Abstract We evaluated a rate colorimetric method (Beckman) for measuring total protein in cerebrospinal fluid. The automated instrument we used was Beckman's ASTRA TM. A 100-microL sample of spinal fluid is introduced into the biuret reagent in the reaction cell and the increase in absorbance at 545 nm is monitored for 20.5 s. Solid-state circuits determine the rate of alkaline biuret-protein chelate formation, which is directly proportional to the total protein concentration in the sample. The linear range of measurement is 120 to 7500 mg/L. Day-to-day precision (CV) over the range of 150 to 1200 mg/L ranged from 15.2 to 2.3%. The method was unaffected by radical alteration of the albumin/globulin ratio, but there is a positive interference in the presence of hemoglobin, a suppression in the presence of bilirubin, and no effect by xanthochromia. The method is precise, accurate, rapid, and convenient. The method was compared with the trichloroacetic acid method as performed on the Du Pont aca III, giving a correlation coefficient (r2) of 0.9693. The method is precise, accurate, rapid, and convenient.

PAROTID SECRETION OF PROTEIN IN MAN

1958 ◽  
Vol 36 (10) ◽  
pp. 1001-1008 ◽  
Author(s):  
Marion H. Ferguson ◽  
H. P. Krahn ◽  
J. A. Hildes
Keyword(s):  
Total Protein ◽  
Protein Concentration ◽  
Serum Proteins ◽  
Amylase Activity ◽  
Early Morning ◽  
Total Protein Concentration ◽  
Zone Electrophoresis ◽  
Residual Radioactivity ◽  
Unstimulated Saliva ◽  
Protein Components

In unstimulated saliva, total protein concentration averaged 186 mg per 100 ml and amylase activity 146 units per 100 ml. The protein concentration was lower in the early morning than at midday. After dilute acetic acid stimulation, both total protein concentration and amylase activity were increased but the concentrations were not affected by rates of secretion above 0.1 ml per minute. Unlike protein, the potassium concentration fell with stimulation.Using zone electrophoresis on filter paper, as many as nine protein components were found, none of which corresponded to the serum proteins. The amylase activity was restricted to a component of low mobility which moved to the anode. There were two or three bands containing glycoproteins; all moved towards the cathode. There were qualitative and quantitative differences between stimulated and unstimulated secretions. Saliva collected 2 or 24 hours after a tracer dose of I131 showed less than 1% residual radioactivity after dialysis or treatment with an anion exchange resin, indicating that little if any of the salivary iodide is organically bound.

Effect of protein concentration on the determination of digoxin in serum by fluorescence polarization immunoassay.

1984 ◽  
Vol 30 (11) ◽  
pp. 1826-1829 ◽  
Author(s):  
W H Porter ◽  
V M Haver ◽  
B A Bush
Keyword(s):  
Serum Protein ◽  
Total Protein ◽  
Fluorescence Polarization ◽  
Protein Concentration ◽  
Total Serum Protein ◽  
Total Serum ◽  
Fluorescence Polarization Immunoassay ◽  
Total Protein Concentration ◽  
Protein Pellet

Abstract Determination of digoxin by fluorescence polarization immunoassay (FPIA) with the Abbott "TDx" is significantly influenced by the concentration of total serum protein. Each 10 g/L increase in serum protein results in an 8% decrease in measured digoxin. Studies with [3H]digoxin confirmed that digoxin binds to the protein pellet during the trichloroacetic acid precipitation step before the immunoassay. Serum protein, or equal concentrations of albumin or gamma-globulin, exert an equivalent effect on the apparent digoxin value. Because the total protein concentration of the assay calibrators is low (50 g/L) compared with its reference interval in serum (60-80 g/L), results by FPIA may be expected to be low by an average of 16% (range, 8-24%). Digoxin results by FPIA will be most nearly accurate when the calibrators include a total protein concentration of about 70 g/L. Patients' specimens with abnormally high or low protein content will give falsely high or low results for digoxin.

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