Interaction of Supramolecular Congo Red and Congo Red-Doxorubicin Complexes with Proteins for Drug Carrier Design

Targeted immunotherapy has expanded to simultaneous delivery of drugs, including chemotherapeutics. The aim of the presented research is to design a new drug carrier system. Systems based on the use of proteins as natural components of the body offer the chance to boost safety and efficacy of targeted drug delivery and excess drug removal. Congo red (CR) type supramolecular, self-assembled ribbon-like structures (SRLS) were previously shown to interact with some proteins, including albumin and antibodies complexed with antigen. CR can intercalate some chemotherapeutics including doxorubicin (Dox). The goal of this work was to describe the CR-Dox complexes, to analyze their interaction with some proteins, and to explain the mechanism of this interaction. In the present experiments, a model system composed of heated immunoglobulin light chain Lλ capable of CR binding was used. Heat aggregated immunoglobulins (HAI) and albumin were chosen as another model system. The results of experiments employing methods such as gel filtration chromatography and dynamic light scattering confirmed the formation of the CR-Dox complex of large size and properties different from the free CR structures. Electrophoresis and chromatography experiments have shown the binding of free CR to heated Lλ while CR-Dox mixed structures were not capable of forming such complexes. HAI was able to bind both free CR and CR-Dox complexes. Albumin also bound both CR and its complex with Dox. Additionally, we observed that albumin-bound CR-Dox complexes were transferred from albumin to HAI upon addition of HAI. DLS analyses showed that interaction of CR with Dox distinctly increased the hydrodynamic diameter of CR-Dox compared with a free CR supramolecular structure. To our knowledge, individual small proteins such as Lλ may bind upon heating a few molecules of Congo red tape penetrating protein body due to the relatively low cohesion of the dye micelle. If, however, the compactness is high (in the case of, e.g., CR-Dox) large ribbon-like, micellar structures appear. They do not divide easily into smaller portions and cannot attach to proteins where there is no room for binding large ligands. Such binding is, however, possible by albumin which is biologically adapted to form complexes with different large ligands and by tightly packed immune complexes and heat aggregated immunoglobulin-specific protein complex structures of even higher affinity for Congo red than albumin. The CR clouds formed around them also bind the CR-Dox complexes. The presented research is essential in the search for optimum solutions for SRLS application in immuno-targeting therapeutic strategies, especially with the use of chemotherapeutics.

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Chitosan Nanocarriers Loading Anti-Tumor Drugs

Journal of Nano Research ◽

10.4028/www.scientific.net/jnanor.32.113 ◽

2015 ◽

Vol 32 ◽

pp. 113-127 ◽

Cited By ~ 2

Author(s):

Ji Wei Wu ◽

Xin Feng Song ◽

Han Wen Sun ◽

Yan Cong Zhang ◽

Xiang Ling Gu ◽

...

Keyword(s):

Self Assembly ◽

Drug Carrier ◽

The Body ◽

Natural Polymer ◽

Preparation Methods ◽

Ionic Crosslinking ◽

Covalent Crosslinking ◽

Immune Ability ◽

Tumor Drugs ◽

Adhesion Effect

Chitosan is a kind of natural polymer commonly applied for nanomaterials, which is affluent in nature with favorable biodegradability and biocompatibility and free of toxicity or odor. In clinic it can be used as a drug carrier for the treatment of cancer, and also it is a kind of new pharmaceutical excipient. To prepare chitosan nanomaterial, various method are used, such as ionic crosslinking, covalent crosslinking, precipitation, free radical polymerization, reverse micelle, spray drying, and self-assembly. Furthermore, plenty of anti-tumor drugs, including adriamycin, epirubicin, taxol, 5-fluorouracil, norcantharidin, folic acid, and so on, are also attempted to load on these chitosan nanocarriers. In addition, the mechanism for those nanocarriers carrying anti-tumor drugs acting on tumor cell were explored, and the formulation mainly include electric charge adhesion effect, suppressing the proliferation of tumor cells, adjusting or enhancing immune ability of the body and inducing apoptosis. This paper compared the characteristics of different preparation methods on chitosan as a nanodrug carrier, summarized the types of packaged drugs, analyzed the mechanism of the chitosan as nanodrug carriers. It can provide valuable reference for researchers' further work.

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THE EFFECT OF FORMALDEHYDE EXPOSURE AND YOGURT SUPPLEMENTATION ON PROFILE AND CHARACTER OF HEPAR TISSUE PROTEIN OF RATS (Rattus norvegicus)

Indonesian Journal of Chemistry ◽

10.22146/ijc.21493 ◽

2010 ◽

Vol 10 (1) ◽

pp. 132-137

Author(s):

Chanif Mahdi ◽

Aulaniam Aulaniam

Keyword(s):

Rattus Norvegicus ◽

Specific Protein ◽

The Body ◽

Oxygen Species ◽

Dot Blot ◽

Laboratory Methods ◽

Carcinogenic Substance ◽

Sds Page ◽

Formaldehyde Exposure ◽

Specific Band

Formaldehyde is a simplest organic compound of aldehyde or alkanal group. Formaldehyde is a toxic and carcinogenic substance. Formaldehyde contamination through food or feeding diet continuously is very dangerous for the body, especially for bodies organ for instances likes hepar and kidney. Because formaldehyde is sources of reactive oxygen species (ROS) and free radicals substances for the body. This purpose of the study is to know the effect of formaldehyde exposure and yogurt supplementation on profile and characters of rats (Rattus norvegicus) protein hepar tissues. The research methods is laboratory methods. The protein profiles determined by electrophoresis (SDS-PAGE) methods. The character of hepar protein tissue determined by ELISA, Dot Blot and Western Blot methods. The result showed that formaldehyde exposure through the feeding diet of rats affect on profile of hepar protein tissue, that characterized by appear of new band of specific protein with molecule weigh is 29.6 kDa (PSForm 29.6). Yogurt supplementation on rat that exposure by formaldehyde through the feeding diet of rat, that characterized by expressing of new band of specific protein with relative molecule weight 24.8 kDa (PSYogh 24.8 kDa), and followed by depressed or dispear of protein specific band of 29.6 kDa(PSForm 29.6 kDa). The result showed that isolated protein PSForm 29.6 kDa have a antigenecity character. Keywords: Formaldehyde exposure, yogurt, ROS, profile and protein character

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Gastruloids develop the three body axes in the absence of extraembryonic tissues and spatially localised signalling

10.1101/104539 ◽

2017 ◽

Cited By ~ 2

Author(s):

D.A. Turner ◽

L. Alonso-Crisostomo ◽

M. Girgin ◽

P. Baillie-Johnson ◽

C. R. Glodowski ◽

...

Keyword(s):

Embryonic Stem ◽

Model System ◽

The Body ◽

Genetic Studies ◽

Animal Development ◽

Embryonic Tissues ◽

Bmp Signalling ◽

Three Body ◽

Extraembryonic Tissues

AbstractEstablishment of the three body axes is a critical step during animal development. In mammals, genetic studies have shown that a combination of precisely deployed signals from extraembryonic tissues position the anteroposterior axis (AP) within the embryo and lead to the emergence of the dorsoventral (DV) and left-right (LR) axes. We have used Gastruloids, embryonic organoids, as a model system to understand this process and find that they are able to develop AP, DV and LR axes as well as to undergo axial elongation in a manner that mirror embryos. The Gastruloids can be grown for 160 hours and form derivatives from ectoderm, mesoderm and endoderm. We focus on the AP axis and show that in the Gastruloids this axis is registered in the expression of T/Bra at one pole that corresponds to the tip of the elongation. We find that localisation of T/Bra expression depends on the combined activities of Wnt/β-Catenin and Nodal/Smad2,3 signalling, and that BMP signalling is dispensable for this process. Furthermore, AP axis specification occurs in the absence of both extraembryonic tissues and of localised sources of signalling. Our experiments show that Nodal, together with Wnt/β-Catenin signalling, is essential for the expression of T/Bra but that Wnt signalling has a separable activity in the elongation of the axis. The results lead us to suggest that, in the embryo, the role of the extraembryonic tissues might not be to induce the axes but to bias an intrinsic ability of the embryo to break its initial symmetry and organise its axes.One sentence summaryCulture of aggregates of defined number of Embryonic Stem cells leads to self-organised embryo-like structures which, in the absence of localised signalling from extra embryonic tissues and under the autonomous influence of Wnt and Nodal signalling, develop the three main axes of the body.

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Plants Extracts Loaded in Nanocarriers: An Emergent Formulating Approach

Natural Product Communications ◽

10.1177/1934578x1801300914 ◽

2018 ◽

Vol 13 (9) ◽

pp. 1934578X1801300 ◽

Cited By ~ 3

Author(s):

Anna Rita Bilia ◽

Vieri Piazzini ◽

Martina Asprea ◽

Laura Risaliti ◽

Giulia Vanti ◽

...

Keyword(s):

Essential Oils ◽

Therapeutic Efficacy ◽

Subcritical Water ◽

Drug Carrier ◽

Drug Loading ◽

The Body ◽

Clinical Use ◽

Intrinsic Dissolution ◽

Optimal Drug ◽

Increasing Demand

Over the millennia, plants have represented for Humankind the main source of food, but also a vast resource to maintain health, for prophylactic properties or to cure human and animal diseases. Presently, between 65 and 80% of populations in developing countries use medicinal plants as therapeutic remedies for their primary healthcare and in Europe and USA there is an increasing demand of botanical products both on the form of food supplements and herbal medicinal products. Botanicals on the market are mainly based on traditional (infusions or decoctions), conventional (using organic solvents) and innovative (supercritical CO2 or subcritical water) extracts but there is an increasing demand of essential oils for aromatherapy. Conversely, the clinical use of many extracts is limited due to the need of repeated administrations or high doses because of low hydrophilicity and intrinsic dissolution rate(s), or physical/ chemical instability. Other limits are low absorption, poor pharmacokinetics and bioavailability, scarce biodistribution, first pass metabolism, trivial penetration and accumulation in the organs of the body. In the case of essential oils, the high volatility and instability are further limitations. Nowadays, the design and production of appropriate drug delivery systems, in particular nanosized ones (between 50 and 300 nm), have already entered into clinical use and can offer an advanced approach to optimized the therapeutic efficacy of extracts and essential oils. A successful drug carrier system should have optimal drug loading and release properties, a long shelf life, and exert a much higher therapeutic efficacy as well as lower side effects. Polymeric nanoparticles and lipid based-nanocarriers including micelles, vesicles, nanocochleates, micro- and nanoemulsions represent successful examples of extract nanoformulations overcoming these limitations. This review reports on some paradigmatic success stories of extract and EO nanoformulations with remarkable advantages over conventional formulations, which include increase of solubility, stability, permeation and bioavailability, sustained delivery. Paradigmatic examples include formulations of extracts from Vitex agnus-castus, Sylibum marianum, Phyllanthus amarus, Ginkgo biloba, Panax notoginseng, Hypericum perforatum and thyme essential oil.

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Nephrocytes are part of the spectrum of filtration epithelial diversity

Cell and Tissue Research ◽

10.1007/s00441-020-03313-7 ◽

2020 ◽

Vol 382 (3) ◽

pp. 609-625

Author(s):

Takayuki Miyaki ◽

Yuto Kawasaki ◽

Akira Matsumoto ◽

Soichiro Kakuta ◽

Tatsuo Sakai ◽

...

Keyword(s):

Body Fluid ◽

Focused Ion Beam ◽

Ion Beam ◽

Three Dimensional ◽

Model System ◽

The Body ◽

Toxic Substances ◽

Cellular Architecture ◽

Animal Phylogeny ◽

Scanning Electron

AbstractThe excretory system produces urine by ultrafiltration via a filtration epithelium. Podocytes are widely found as filtration epithelial cells in eucoelomates. In some animal taxa, including insects and crustaceans, nephrocytes serve to separate toxic substances from the body fluid, in addition to podocytes. Drosophila nephrocytes have been recently utilized as a model system to study podocyte function and disease. However, functionality and cellular architecture are strikingly different between Drosophila nephrocytes and eucoelomate podocytes, and the phylogenetic relationship between these cells remains enigmatic. In this study, using focused-ion beam-scanning electron microscopy (FIB-SEM) tomography, we revealed three-dimensional architecture of decapod nephrocytes with unprecedented accuracy—they filled an enormous gap, which can be called “missing link,” in the evolutionary diversity of podocytes and nephrocytes. Thus, we concluded that nephrocytes are part of the spectrum of filtration epithelial diversity in animal phylogeny.

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Setaria cervidual specific phosphatase: characterization and its effect on eosinophil degranulation

Parasitology ◽

10.1017/s0031182009006271 ◽

2009 ◽

Vol 136 (8) ◽

pp. 895-904 ◽

Cited By ~ 5

Author(s):

S. RATHAUR ◽

R. RAI ◽

E. SRIKANTH ◽

S. SRIVASTAVA

Keyword(s):

Protein Tyrosine Phosphatase ◽

Tyrosine Phosphatase ◽

Gel Filtration ◽

Specific Protein ◽

Enzyme Concentration ◽

Cross Reactivity ◽

Filarial Parasite ◽

Significant Activity ◽

Protein Tyrosine

SUMMARYSetaria cervi, a bovine filarial parasite contains significant acid phosphatase (AcP) activity in its various life stages. Two forms of AcP were separated from somatic extract of adult female parasite using cation exchange, gel filtration and concavalin affinity chromatography. One form having a molecular mass of 79 kDa was characterized as dual specific protein tyrosine phosphatase (ScDSP) based on substrate specificity and inhibition studies. With various substrates tested, it showed significant activity in the order of phospho-L-tyrosine>pNPP>ADP>phospho-L-serine. Inhibition by orthovanadate, fluoride, molybdate, and zinc ions further confirms protein tyrosine phosphatase nature of the enzyme. Km and Vmax determined with various substrates were found to be 16·66 mM, 25·0 μM/ml/min with pNPP; 20·0 mM, 40·0 μM/ml/min with phospho-L-tyrosine and 27·0 mM, 25·0 μM/ml/min with phospho-L-serine. KIwith pNPP and sodium orthovanadate (IC5033·0 μM) was calculated to be 50·0 mM. Inhibition with pHMB, silver nitrate, DEPC and EDAC suggested the presence of cysteine, histidine and carboxylate residues at its active site. Cross-reactivity withW. bancrofti-infected sera was demonstrated by Western blotting. ScDSP showed elevated levels of IgE in chronic filarial sera using ELISA. Underin vitroconditions, ScDSP resulted in increased effector function of human eosinophils when stimulated by IgG, which showed a further decrease with increasing enzyme concentration. Results presented here suggest thatS. cerviDSP should be further studied to determine its role in pathogenesis and the persistence of filarial parasite.

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Biotin-binding protein from chicken egg yolk. Assay and relationship to egg-white avidin

Biochemical Journal ◽

10.1042/bj1570395 ◽

1976 ◽

Vol 157 (2) ◽

pp. 395-400 ◽

Cited By ~ 38

Author(s):

H B White ◽

B A Dennison ◽

M A Della Fera ◽

C J Whitney ◽

J C McGuire ◽

...

Keyword(s):

Binding Protein ◽

Gel Filtration ◽

Egg Yolk ◽

Egg White ◽

Specific Protein ◽

Chicken Egg ◽

Biotin Binding ◽

Lower Temperature ◽

Chicken Egg Yolk ◽

Kinetics Of

1. Biotin in chicken egg yolk is non-covalently bound to a specific protein that comprises 0.03% of the total yolk protein (0.8 mg/yolk). This biotin-binding protein is not detectable by the normal avidin assay owing to the biotin being tightly bound. Exchange of [14C]biotin for bound biotin at 65 degrees C is the basis of an assay for this protein. 2. Biotin-binding protein from egg yolk is distinguishable from egg-white avidin on Sephadex G-100 gel filtration, although the sizes of the two proteins appear quite similar. 3. Biotin-binding protein is denatured at a lower temperature and freely exchanges biotin at lower temperatures than does avidin. 4. The biotin-binding protein in egg yolk is postulated to be responsible for the deposition of biotin in egg yolk. D-[carboxyl-14C]Biotin injected into laying hens rapidly appears in the egg bound to yolk biotin-binding protein and avidin. Over 60% of the radioactivity is eventually deposited in eggs. The kinetics of biotin deposition in the egg suggests a 25 day half-life for an intracellular biotinyl-coenzyme pool in the laying hen.

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Isolation and identification of metallothionein isoforms (MT-1 and MT-2) in the rat testis

Biochemical Journal ◽

10.1042/bj3340695 ◽

1998 ◽

Vol 334 (3) ◽

pp. 695-701 ◽

Cited By ~ 29

Author(s):

Junko S. SUZUKI ◽

Naomi KODAMA ◽

Andrei MOLOTKOV ◽

Etsuko AOKI ◽

Chiharu TOHYAMA

Keyword(s):

Metal Binding ◽

Binding Proteins ◽

Gel Filtration ◽

Specific Protein ◽

Composition Analysis ◽

Isolation And Identification ◽

Rat Testis ◽

Heavy Metal Binding ◽

Protein Fragmentation ◽

Male Genital

It has been a long-lasting controversial issue as to whether or not the male genital organs, such as the testis and prostate, contain metallothioneins (MTs), a group of cysteine-rich heavy-metal-binding proteins that play a role in detoxifying heavy metals such as cadmium (Cd). Earlier studies reported that the rodent testis lacks MTs and concluded that this is why the testis is very susceptible to Cd, although other indirect experimental evidence suggests that MTs are present in this organ. A deficiency of MTs in the testis was originally suspected on the basis of amino acid composition analysis, since MT-like proteins isolated as Cd-binding proteins did not have a characteristic MT structure. In the present study, we demonstrate that the rat testis indeed expresses Cd-binding proteins with sequences identical to those previously described for MT-1 and MT-2, the major isoforms. To confirm that MT-1 and MT-2 are present in the rat testis, we purified and isolated Cd-binding proteins by homogenization using Cd-containing buffer, followed by sequential purification using Sephadex G-75 gel filtration chromatography and anion HPLC column chromatography, which yielded Cd-binding protein-1 (Cd-BP-1) and -2 (Cd-BP-2). After pyridylethylation, Cd-BP-1 and Cd-BP-2 were subjected to specific protein fragmentation by acids and endopeptidases, which revealed that these Cd-binding proteins have the same primary structures as MT-1 and MT-2 respectively. Thus we believe that the present results clearly resolve the long-standing debate about the presence of MTs in the testis, at least in the rodent.

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Nutritional status and intake regulation in sheep. VIII.* Relationships between voluntary intake of herbage by sheep and the protein/energy ratio in the digestion products

Australian Journal of Agricultural Research ◽

10.1071/ar9770907 ◽

1977 ◽

Vol 28 (5) ◽

pp. 907 ◽

Cited By ~ 53

Author(s):

AR Egan

Keyword(s):

Energy Intake ◽

Oral Intake ◽

Specific Protein ◽

The Body ◽

Energy Ratio ◽

Protein Energy ◽

Energy Ratios ◽

Digestible Protein ◽

Voluntary Intake ◽

Intake Regulation

In data from two separate experiments in which the same herbage diets were fed to sheep, a relationship was observed between the protein/energy ratio in digestion products and the level of voluntary feed intake: I = 0.16P—0.16 (SEb = 0.015; r2 = 0.85), where I is the voluntary intake of digestible energy (DE) (MJ/W0.75), P the protein digested in the intestine (g/MJ DE) and W the body weight (kg). When supplementary casein was infused into the duodenum of sheep fed on 15 basal diets, intake changes were greatest (up to 15% increase) with six roughage diets, in which estimated truly digestible protein contributed 5.5 g digested protein (DP) per MJ DE (about 10% of DE as protein) or less. No responses were observed with two other roughages in the same range or with seven roughages for which the estimated truly digestible protein contributed more than 6 g per MJ DE (about 13% of DE as protein). The change in voluntary intake was not found to be simply linked to the protein input, in that a consistent overall estimated protein/energy ratio in digestion products was not established as voluntary intake changed in response to protein infusion. The estimated resultant protein/energy ratios established were always high (7.4–9.4 g DP/MJ DE) relative to those observed on the basal diets (3.4–8.4 g DP/MJ DE). In a further experiment with a wheat hay–straw diet, voluntary intake was measured during periods of infusion of acetic acid per rumen, and/or protein (casein) infusion per duodenum. Energy infusion and protein infusion could be shown qualitatively to have opposed effects on oral intake. However, oral intake adjustments did not appear to act to preserve or re-establish any specific Protein/energy ratio in the total nutrients absorbed. The observations are discussed in relation to factors controlling energy intake, and the effect of protein inadequacy upon level of energy intake in the sheep. *Part VII, Aust. J. Agric. Res., 23: 247 (1972).

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Insulin-like growth factors and their binding proteins in pleural fluid

Acta Endocrinologica ◽

10.1530/eje.0.1370467 ◽

1997 ◽

pp. 467-473 ◽

Cited By ~ 4

Author(s):

Y Le Bouc ◽

A Bellocq ◽

C Philippe ◽

L Perin ◽

M Garabedian ◽

...

Keyword(s):

Growth Factors ◽

Pleural Fluid ◽

Binding Proteins ◽

Specific Binding ◽

Gel Filtration ◽

Specific Protein ◽

Hydroxyvitamin D ◽

Igf I ◽

Acid Gel ◽

Insulin Like Growth Factors

We investigated the expression and potential regulatory role of insulin-like growth factors (IGFs) and their specific binding proteins (BPs) in tuberculous and nontuberculous pleuritis. By using a radioimmunoassay after acid gel filtration chromatography, we found that mean concentrations of IGF-I were 211.9 +/- 20.2 microg/l and 203.2 +/- 31.1 microg/l in pleural fluid of 14 patients with tuberculous pleuritis and 9 patients with malignant pleuritis respectively. These values were near those in serum of the same patients (221.3 +/- 19.5 microg/l and 204.6 +/- 21.0 microg/l respectively). By using a specific protein-binding assay, we found that mean concentrations of IGF-II were 345.3 +/- 61.0 microg/l and 167.6 +/- 22.7 microg/l in tuberculous and malignant pleural effusions respectively. These values were significantly lower than those in serum of the same patients (628.3 +/- 79.0 microg/l, P<0.025 and 532.0 +/- 85.9 microg/l, P<0.025 respectively). Because bioavailability and bioactivity of IGFs may be regulated by their binding to IGFBPs, we studied IGFBP patterns in the pleural fluid of 6 patients with tuberculous pleuritis. As assessed by Western ligand blotting the levels of IGFBP-1 and IGFBP-2 were increased whereas those of IGFBP-3 were decreased in pleural fluid in comparison with serum. The decrease in IGFPB-3 levels reflected increased proteolysis, as assessed by Western immunoblotting. In spite of this presence of IGFBPs, IGFs could be responsible for the local biosynthesis of 1.25-dihydroxyvitamin D (1,25-(OH)2D) since pleural fluid levels of both IGF-I and IGF-II significantly correlated with those of 1,25-(OH)2D. These results indicate that IGFs are detectable in pleural fluid and may contribute to control the activity of 25-hydroxyvitamin D-1alpha hydroxylase in tuberculous pleuritis.

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