scholarly journals Protein-bound polyphenols create “ghost” band artifacts during chemiluminescence-based antigen detection

F1000Research ◽  
2017 ◽  
Vol 6 ◽  
pp. 254
Author(s):  
Nathalie Plundrich ◽  
Mary Ann Lila ◽  
Edward Foegeding ◽  
Scott Laster
Keyword(s):  
Horseradish Peroxidase ◽  
Green Tea ◽  
Western Blotting ◽  
Specific Ige ◽  
Egg White ◽  
Antigen Detection ◽  
Band Formation ◽  
Western Blots ◽  
Horseradish Peroxide ◽  
The Impact

Antigen detection during Western blotting commonly utilizes a horseradish peroxidase-coupled secondary antibody and enhanced chemiluminescent substrate. We utilized this technique to examine the impact of green tea-derived polyphenols on the binding of egg white protein-specific IgE antibodies from allergic human plasma to their cognate antigens. Our experiments unexpectedly showed that green tea-derived polyphenols, when stably complexed with egg white proteins, caused “ghost” band formation in the presence of horseradish peroxide. This study suggests that caution should be taken when evaluating polyphenol-bound proteins by enhanced chemiluminescence Western blotting using horseradish peroxidase and demonstrates that protein-bound polyphenols can be a source of “ghost” band artifacts on Western blots.

scholarly journals Protein-bound polyphenols create “ghost” band artifacts during chemiluminescence-based antigen detection

F1000Research ◽  
2017 ◽  
Vol 6 ◽  
pp. 254 ◽  
Author(s):  
Nathalie Plundrich ◽  
Mary Ann Lila ◽  
Edward Foegeding ◽  
Scott Laster
Keyword(s):  
Horseradish Peroxidase ◽  
Green Tea ◽  
Western Blotting ◽  
Specific Ige ◽  
Egg White ◽  
Antigen Detection ◽  
Western Blots ◽  
Enhanced Chemiluminescence ◽  
Specific Ige Antibodies ◽  
The Impact

Antigen detection during Western blotting commonly utilizes a horseradish peroxidase-coupled secondary antibody and enhanced chemiluminescent substrate. We utilized this technique to examine the impact of green tea-derived polyphenols on the binding of egg white protein-specific IgE antibodies from allergic human plasma to their cognate antigens. Our experiments unexpectedly showed that green tea-derived polyphenols, when stably complexed with egg white proteins, caused hyperactivation of horseradish peroxidase resulting in the appearance of white “ghost” bands. This study suggests that caution should be taken when evaluating polyphenol-bound proteins by enhanced chemiluminescence Western blotting using horseradish peroxidase and demonstrates that protein-bound polyphenols can be a source of “ghost” band artifacts on Western blots.

Pengaruh Lama Pengasinan Terhadap Kadar Protein Putih Telur Itik

2014 ◽  
Vol 1 (1) ◽  
pp. 36 ◽  
Author(s):  
Siti Fatimah ◽  
Muji Rahayu ◽  
Siti Aminah
Keyword(s):  
Protein Level ◽  
Egg White ◽  
Special Treatment ◽  
Egg White Protein ◽  
Protein Levels ◽  
Duck Egg ◽  
Egg Period ◽  
Graph Presentation ◽  
And Control ◽  
The Impact

Background : Egg  is one of the animal protein source, which has delicious taste, easy to digest and highly nutritious. Besides its affordable price, its supply availability is unquestionable as well. However, due to its short storability, it requires special treatment, such as preserving, to store it for long period. One way to preserve the egg is by pickling egg, which generally requires seven to ten days of marinating. During the process of marinating, there will be a visual change of egg white and yolk. Their structures  will be more solid (the occurrence of thickening process) because salinization will lead to protein denaturalization. Consequently, it has an influence as well towards the content of egg white protein of duck egg. This study is aimed to explore the impact of various time of pickling egg towards egg white protein of duck egg. Method  : The study where takes place in a laboratories, is a true experimental study for the reason that the researcher must provide intervention, hence all of potentially confounding variables are manageable. Samples that had been used in this study are duck eggs which were bought from North Brebes. This study is expected to generate data from four various time of pickling egg and control (no treatment). Since there are four samples, accordingly the number of data resulted are twenty. The resulted data will be descriptively presented in table, graph, presentation, and narration. Result  : Protein level examination within duck white egg shows changes  in protein levels that occurs in every variation of pickling egg time, where the average results of the assay of duck egg white protein is 14.94% without treatment (control), in five days of pickling time is 13.68%, in seven days of pickling time is 13.29%, in nine days of pickling time is 12.87% and eleven days of pickling time is 12.78%. Conclusion  : There is a significant impact among the period of pickling time to the protein level degradation of duck white egg. Keywords : Duck egg, period of pickling time, level protein of duck white egg.

scholarly journals The Importance of Diet in Predicting the Remission of Urticaria—Determination of Allergen-Specific IgE

Medicina ◽  
2021 ◽  
Vol 57 (7) ◽  
pp. 679
Author(s):  
Monica Iuliana Ungureanu ◽  
Liliana Sachelarie ◽  
Radu Ciorap ◽  
Bogdan Aurelian Stana ◽  
Irina Croitoru ◽  
...  
Keyword(s):  
Predictive Value ◽  
Allergic Diseases ◽  
Specific Ige ◽  
Egg White ◽  
Food Allergens ◽  
Pediatric Hospital ◽  
Disease Remission ◽  
Specific Serum ◽  
Different Types

Background and Objectives: Different types of food introduced gradually in the diet will expose children to different food allergens, increasing the chance of developing allergic diseases. The aim of our study was to determine if allergen-specific IgE values can influence, depending on the diet, the prediction of remission of urticaria in children. Materials and Methods: This prospective study was conducted in 132 patients diagnosed over two years with urticaria, admitted to “Sf. Maria” Clinical Pediatric Hospital Iaşi. Total IgE assay was performed by ELISA, and determination of specific serum IgE by the CLA System Quanti Scan method (Innogenetics, Heiden, Germany). Data were gathered and statistical analysis was performed using statistical software SPSS, using descriptive and inferential statistics. Results: The determination of specific IgE to food allergens was performed on a total of 132 cases. The values of specific IgE were positive for one or more food allergens in 84 patients (63.64%). The most common allergens involved were: cow’s milk in 33.3% cases, egg white in 22.6% cases, and hazelnuts in 11.9% cases. The specific IgE values for the different types of food included in our study had a predictive value for disease remission. Conclusions: The determination of specific IgE confirms the presence of a particular food allergen and may have predictive value for the future development of an allergic manifestation.

scholarly journals Ingredient-Dependent Extent of Lipid Oxidation in Margarine

Antioxidants ◽  
2021 ◽  
Vol 10 (1) ◽  
pp. 105
Author(s):  
Sarah Fruehwirth ◽  
Sandra Egger ◽  
Dennis Kurzbach ◽  
Jakob Windisch ◽  
Franz Jirsa ◽  
...  
Keyword(s):  
Citric Acid ◽  
Lipid Oxidation ◽  
Oxidative Stability ◽  
Green Tea ◽  
Acyl Chain ◽  
Green Tea Extract ◽  
Tocopheryl Acetate ◽  
Tea Extract ◽  
The Impact ◽  
Β Carotene

This study reports the impact of margarine-representative ingredients on its oxidative stability and green tea extract as a promising antioxidant in margarine. Oil-in-water emulsions received much attention regarding factors that influence their oxidative stability, however, water-in-oil emulsions have only been scarcely investigated. Margarine, a widely consumed water-in-oil emulsion, consists of 80–90% fat and is thermally treated when used for baking. As different types of margarine contain varying additives, their impact on the oxidative stability of margarine during processing is of pressing importance. Thus, the influence of different ingredients, such as emulsifiers, antioxidants, citric acid, β-carotene and NaCl on the oxidative stability of margarine, heated at 80 °C for 1 h to accelerate lipid oxidation, was analyzed by the peroxide value and oxidation induction time. We found that monoglycerides influenced lipid oxidation depending on their fatty acyl chain. α-Tocopheryl acetate promoted lipid oxidation, while rosemary and green tea extract led to the opposite. Whereas green tea extract alone showed the most prominent antioxidant effect, combinations of green tea extract with citric acid, β-carotene or NaCl increased lipid oxidation in margarine. Complementary, NMR data suggested that polyphenols in green tea extracts might decrease lipid mobility at the surface of the water droplets, which might lead to chelating of transition metals at the interface and decreasing lipid oxidation.

scholarly journals miR-208b Reduces the Expression of Kcnj5 in a Cardiomyocyte Cell Line

Biomedicines ◽  
2021 ◽  
Vol 9 (7) ◽  
pp. 719
Author(s):  
Julia Hupfeld ◽  
Maximilian Ernst ◽  
Maria Knyrim ◽  
Stephanie Binas ◽  
Udo Kloeckner ◽  
...  
Keyword(s):  
Untranslated Region ◽  
Electrical Remodeling ◽  
Sequencing Data ◽  
Western Blots ◽  
Cardiovascular Pathology ◽  
Hypertrophic Response ◽  
Channel Dependent ◽  
Cardiac Excitation ◽  
The Impact ◽  
Generation Sequencing

MicroRNAs (miRs) contribute to different aspects of cardiovascular pathology, among them cardiac hypertrophy and atrial fibrillation. Cardiac miR expression was analyzed in a mouse model with structural and electrical remodeling. Next-generation sequencing revealed that miR-208b-3p was ~25-fold upregulated. Therefore, the aim of our study was to evaluate the impact of miR-208b on cardiac protein expression. First, an undirected approach comparing whole RNA sequencing data to miR-walk 2.0 miR-208b 3′-UTR targets revealed 58 potential targets of miR-208b being regulated. We were able to show that miR-208b mimics bind to the 3′ untranslated region (UTR) of voltage-gated calcium channel subunit alpha1 C and Kcnj5, two predicted targets of miR-208b. Additionally, we demonstrated that miR-208b mimics reduce GIRK1/4 channel-dependent thallium ion flux in HL-1 cells. In a second undirected approach we performed mass spectrometry to identify the potential targets of miR-208b. We identified 40 potential targets by comparison to miR-walk 2.0 3′-UTR, 5′-UTR and CDS targets. Among those targets, Rock2 and Ran were upregulated in Western blots of HL-1 cells by miR-208b mimics. In summary, miR-208b targets the mRNAs of proteins involved in the generation of cardiac excitation and propagation, as well as of proteins involved in RNA translocation (Ran) and cardiac hypertrophic response (Rock2).

scholarly journals Localization of Na+,K+-ATPase alpha-subunit to the sinusoidal and lateral but not canalicular membranes of rat hepatocytes.

1987 ◽  
Vol 104 (5) ◽  
pp. 1239-1248 ◽  
Author(s):  
E S Sztul ◽  
D Biemesderfer ◽  
M J Caplan ◽  
M Kashgarian ◽  
J L Boyer
Keyword(s):  
Monoclonal Antibody ◽  
Plasma Membrane ◽  
Horseradish Peroxidase ◽  
Rat Liver ◽  
Plasma Membranes ◽  
Alpha Subunit ◽  
Gamma Glutamyl Transferase ◽  
Surface Distribution ◽  
Western Blots ◽  
Glutamyl Transferase

Controversy has recently developed over the surface distribution of Na+,K+-ATPase in hepatic parenchymal cells. We have reexamined this issue using several independent techniques. A monoclonal antibody specific for the endodomain of alpha-subunit was used to examine Na+,K+-ATPase distribution at the light and electron microscope levels. When cryostat sections of rat liver were incubated with the monoclonal antibody, followed by either rhodamine or horseradish peroxidase-conjugated goat anti-mouse secondary, fluorescent staining or horseradish peroxidase reaction product was observed at the basolateral surfaces of hepatocytes from the space of Disse to the tight junctions bordering bile canaliculi. No labeling of the canalicular plasma membrane was detected. In contrast, when hepatocytes were dissociated by collagenase digestion, Na+,K+-ATPase alpha-subunit was localized to the entire plasma membrane. Na+,K+-ATPase was quantitated in isolated rat liver plasma membrane fractions by Western blots using a polyclonal antibody against Na+,K+-ATPase alpha-subunit. Plasma membranes from the basolateral domain of hepatocytes possessed essentially all of the cell's estimated Na+,K+-ATPase catalytic activity and contained a 96-kD alpha-subunit band. Canalicular plasma membrane fractions, defined by their enrichment in alkaline phosphatase, 5' nucleotidase, gamma-glutamyl transferase, and leucine aminopeptidase had no detectable Na+,K+-ATPase activity and no alpha-subunit band could be detected in Western blots of these fractions. We conclude that Na+,K+-ATPase is limited to the sinusoidal and lateral domains of hepatocyte plasma membrane in intact liver. This basolateral distribution is consistent with its topology in other ion-transporting epithelia.

scholarly journals The Impact of Green Tea and Coffee Consumption on the Reduced Risk of Stroke Incidence in Japanese Population

Stroke ◽  
2013 ◽  
Vol 44 (5) ◽  
pp. 1369-1374 ◽  
Author(s):  
Yoshihiro Kokubo ◽  
Hiroyasu Iso ◽  
Isao Saito ◽  
Kazumasa Yamagishi ◽  
Hiroshi Yatsuya ◽  
...  
Keyword(s):  
Green Tea ◽  
Japanese Population ◽  
Coffee Consumption ◽  
Stroke Incidence ◽  
The Impact ◽  
Reduced Risk

scholarly journals Component resolved diagnosis of egg yolk is an indispensable part of egg allergy

2021 ◽  
Vol 49 (2) ◽  
pp. 6-14
Author(s):  
Huang Lunhui ◽  
Shao Yanhong ◽  
Li Shaoshen ◽  
Bao Huijing ◽  
Liu Yunde ◽  
...  
Keyword(s):  
Operating Characteristic ◽  
Egg Yolk ◽  
Specific Ige ◽  
Diagnostic Value ◽  
Egg White ◽  
Enzyme Linked Immunosorbent Assay ◽  
Egg Allergy ◽  
Component Resolved Diagnosis ◽  
Allergen Components

Introduction and objectives: It was urgent to explain the role of egg yolk allergen sensitization to the egg allergic population and we would evaluate the diagnostic value of allergen components in whole eggs, including egg white and egg yolk.Materials and methods: Firstly, we collected 99 positive and 21 negative sera against egg allergy. Then we used modified enzyme linked immunosorbent assay (ELISA) to survey specific IgE (sIgE) to all-proven and single component in eggs, Ovomucoid (Gal d 1), Ovalbumin (Gal d 2), Ovotransferrin (Gal d 3), Lysozyme C (Gal d 4), Serum Albumin (Gal d 5), and YGP42(Gal d 6) in allergic and non-allergic populations. Last but not least, we studied the sIgE reactivities to egg allergen components by receiver operating characteristic (ROC) analysis.Results: Among egg-allergic individuals, nearly 10% were sensitized to five of six egg allergen components, and the cross-reaction frequency between two egg yolk allergens with Gal d 1 was about 30% in the groups diagnosed with egg allergy or non-allergy. The best component-combination diagnosis in egg allergy of Gal d 1+ Gal d 6 demonstrated the largest area under curve (AUC) of 0.994.Conclusions: Our results suggested that there were individual differences in allergenicity of different egg allergen components, especially in the samples negative to egg allergy diagnosed but sensitive to egg yolk components. It was indicated that component resolved diagnosis of egg yolk improved the value for egg allergy management indispensably.

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