Dynamics of absorption of C-14-biotin by blood cells and plasma protein binding in ontogenesis

Age peculiarities of absorption of 14C-biotin by the blood cells and plasma proteins in vitro are studied. The phase dynamics of 14C-biotin absorption by the blood cells and binding by plasma proteins is found. The reliable ontogenetic differences in absorption of 14C-biotin by the blood cells and plasma proteins are determined. The biotin-binding in vitro by the blood cells and plasma proteins decreases and becomes slower in old rats as compared with young ones.     

THE ROUTE OF ENTRY AND LOCALIZATION OF BLOOD PROTEINS IN THE OOCYTES OF SATURNIID MOTHS

The oocytes of saturniid moths take up proteins selectively from the blood. The distribution of blood proteins in the ovary during protein uptake was investigated by staining 2 µ sections of freeze-dried ovaries with fluorescein-labeled antibodies. The results indicate that blood proteins occur primarily in the intercellular spaces of the follicle cell layer, in association with a brush border at the surface of the oocyte, and within the oocyte in the yolk spheres. That proteins derived from the blood are associated with the yolk spheres was confirmed by isolating these bodies and showing that lysis, which can be induced by any of a number of mechanical means, causes them to release immunologically defined proteins known to be derived from the blood. That the level of blood proteins in the cytoplasm is low relatively to that in the yolk spheres was confirmed by the observation that the yellow pigments associated with several blood proteins, although conspicuous in the yolk spheres, are not visible in the translucent layer of centrifuged oocytes. From these and previous physiological observations, it is proposed that blood proteins reach the surface of the oocyte by an intercellular route, that they combine with some component of the brush border, and that they are transformed into yolk spheres by a process akin to pinocytosis.     

Sorption of plasma proteins by fluorocarbon emulsions stabilized by various surfactants

It has been found in in vivo and in vitro experiments that, as a perfluorocarbon emulsion stabilized by Proxanol 268 comes in contact with blood plasma proteins, plasma proteins with molecular masses from 25 to 170 kDa and above are adsorbed on the surface of emulsion particles. Among the adsorbed proteins, fibronectin and fibrinogen were identified by immunoblotting. In in vivo experiments, during circulation in the blood flow, considerable amounts of plasma proteins are adsorbed on Proxanol-stabilized emulsion particles; the amount of adsorbed proteins increases with the time the particles are in the blood flow. Considerably lesser amounts of proteins are adsorbed during circulation in the blood flow on emulsion particles stabilized by egg yolk phospholipids, and their qualitative composition differs from the composition of proteins adsorbed on Proxanol-stabilized emulsion particles. A preliminary incubation of the Proxanol-stabilized emulsion with heparin decreases the amount of the adsorbed proteins and changes their qualitative composition.     

Hypotensive activity of the plasma proteins in a normovolemic blood exchange transfusion

The effect of post-hemotransfusion protein fractions on blood pressure, microcirculation and physiologically active substances has been studied in stimulated blood replacement by homologous animal blood. The in vivo and in vitro experiments have revealed that subfraction of hemotransfusion plasma macromolecular proteins has a prominent antihypertensive effect, leading to blood flow slowing in the microvascular bed. Hemotransfusion plasma proteins possess high serotonin-releasing activity. The involvement of blood proteins and physiologically active substances into the generation of the recepient's response to homologous blood transfusion from several donors and its role in the genesis of post-transfusion complications are discussed.     

Sorption of plasma components on the surface of particles of fluorocarbon emulsions stabilized by proxanol 268

It was found in the experiments in vivo and in vitro that the contact of perfluorocarbon emulsion stabilized with proxanol 268 with blood plasma leads to the sorption of various plasma proteins on the surface of emulsion particles. The profile of the proteins sorbed is complex and includes proteins with molecular weights ranging from 14 to 94 kDa. Among proteins sorbed on the emulsion particles circulating in blood, IgG was identified. Incubation of the emulsion stabilized with proxanol 268 with human blood plasma in vitro was shown to result in the sorption of IgG and IgA the perfluorocarbon particles. The sorbtion of serum proteins and immune complexes circulating in blood on the surface of perfluorocarbon particles stabilized with proxanol 268 was revealed to activate the complement system.     

Vascular wall extracellular matrix proteins and vascular diseases

Extracellular matrix proteins form the basic structure of blood vessels. Along with providing basic structural support to blood vessels, matrix proteins interact with different sets of vascular cells via cell surface integrin or non-integrin receptors. Such interactions induce vascular cell de novo synthesis of new matrix proteins during blood vessel development or remodeling. Under pathological conditions, vascular matrix proteins undergo proteolytic processing, yielding bioactive fragments to influence vascular wall matrix remodeling. Vascular cells also produce alternatively spliced variants that induce vascular cell production of different matrix proteins to interrupt matrix homeostasis, leading to increased blood vessel stiffness; vascular cell migration, proliferation, or death; or vascular wall leakage and rupture. Destruction of vascular matrix proteins leads to vascular cell or blood-borne leukocyte accumulation, proliferation, and neointima formation within the vascular wall; blood vessels prone to uncontrolled enlargement during blood flow diastole; tortuous vein development; and neovascularization from existing pathological tissue microvessels. Here we summarize discoveries related to blood vessel matrix proteins within the past decade from basic and clinical studies in humans and animals — from expression to cross-linking, assembly, and degradation under physiological and vascular pathological conditions, including atherosclerosis, aortic aneurysms, varicose veins, and hypertension.     

Radioimmunological determination of antibodies of DNA and other DNA-binding proteins in the blood serum of immunized animals

The DNA-binding activity of blood serum proteins was determined in the process of the immune response accompanied and not accompanied by the appearance of antibodies to DNA in blood immunized animals. The immunological rearrangements in the organism following DNA administration without the appearance of antibodies against a unihelical DNA in the blood serum cause an increase in the DNA-binding ability of blood serum proteins, which decreases the specificity of the radioimmunological method in determination of antibodies to DNA. Denaturation of DNA in the presence of formalin also increases the nonspecific binding with DNA of blood serum proteins containing and noncontaining antibodies to DNA.     

14C] Alanine incorporation into proteins and their contents in the tissue of intact and adrenalectomized rabbits after hydrocortisone and cortitropin administration

A single administration of hydrocortisone to intact rabbits increases the incorporation of [14C] alanine into proteins of the brain and liver tissue homogenates and soluble fractions as well as in blood plasma proteins and reduces the label incorporation into the brain tissue proteins and reduces its incorporation into the blood plasma proteins. Adrenalcetomy is followed by an increase in the incorporation of [14C] alanine into proteins of the brain and muscle tissue homogenates and soluble fraction and into proteins of blood plasma and liver tissue homogenates as well as by reducing the label incorporation into the spleen soluble fraction proteins. ACTH administered to adrenalectomized rabbits reduces incorporation of [14C] alanine into the brain and muscle tissue proteins, total proteins of liver tissue homogenate and increases it into the proteins of the spleen tissue soluble fraction. Multiple administration of the soluble fraction hormones both to intact and adrenalectomized rabbits inhibits the label incorporation into the studied tissue proteins. Parallel with the change in [14C] alanine incorporation into proteins under the hormones effect certain shifts in their contents were also established.     

Distribution of porphyrin sensitizers among protein and cellular blood elements

The distribution of porphyrin pigments between plasma proteins and blood cells was studied. It was shown that the relative fraction of sensitizer bound by blood cells changed significantly depending on the physicochemical features of pigment molecules. This parameter strongly correlates with porphyrin polarity. Polar watersoluble tetraphenylporphin derivatives, chlorine e6 and hematoporphyrin are bound by plasma proteins only. The decrease in pigment polarity by substitution of polar side groups results in a drastic increase of pigment affinity to blood cells. The binding of extremely apolar pigments by cells in blood occurs for a long period of time, probably as a result of a low rate of pigment redistribution between serum proteins and cellular membrane. The data obtained show that blood cells may be involved into the control of pigment transport and distribution in organism during photodynamic therapy. The parameters of porphyrin distribution between plasma proteins and cells in blood are of certain importance when the pharmacokinetic behavior of various sensitizers is compared.     

The impact of blood contamination on the proteome of cerebrospinal fluid

Human cerebrospinal fluid (CSF) is in direct contact with the brain extracellular space. Beside the secretion of CSF by the choroid plexus the fluid also derives directly from the brain by the ependymal lining of the ventricular system and the glial membrane and from blood vessels in the arachnoid. Therefore, biochemical change in the brain may be reflected in the CSF. CSF is a potential source of protein molecular indices of central nervous system function and pathology. However, various amounts of blood contamination in CSF may arise during sample acquisition. The concentration of protein in the CSF is only 0.2 to 0.5% that of blood. Minor contamination of CSF with blood during collection of the fluid may dramatically alter the protein profile confounding the identification of potential biomarkers. We have analyzed CSF and CSF spiked with increasing amounts of whole blood using proteomic techniques. We detected at least four blood specific highly abundant proteins: hemoglobin, catalase, peroxiredoxin and carbonic anhydrase I. These proteins can be used as blood contamination markers for proteomic analysis of CSF. Proteins in blood contaminated CSF samples were less stable compared to neat CSF at 37 degrees C suggesting that blood borne protease may induce protein degradation in CSF during sample acquisition. This analysis was aimed at identification of proteins found primarily in CSF, those found primarily in blood and assessment of the impact of blood contamination on those proteins found in both fluids.     

The human erythrocyte proteome: analysis by ion trap mass spectrometry

This report describes an analysis of the red blood cell proteome by ion trap tandem mass spectrometry in line with liquid chromatography. Mature red blood cells lack all internal cell structures and consist of cytoplasm within a plasma membrane envelope. To maximize outcome, total red blood cell protein was divided into two fractions of membrane-associated proteins and cytoplasmic proteins. Both fractions were divided into subfractions, and proteins were identified in each fraction separately through tryptic digestion. Membrane protein digests were collected from externally exposed proteins, internally exposed proteins, "spectrin extract" mainly consisting of membrane skeleton proteins, and membrane proteins minus spectrin extract. Cytoplasmic proteins were divided into 21 fractions based on molecular mass by size exclusion chromatography. The tryptic peptides were separated by reverse-phase high-performance liquid chromatography and identified by ion trap tandem mass spectrometry. A total of 181 unique protein sequences were identified: 91 in the membrane fractions and 91 in the cytoplasmic fractions. Glyceraldehyde-3-phosphate dehydrogenase was identified with high sequence coverage in both membrane and cytoplasmic fractions. Identified proteins include membrane skeletal proteins, metabolic enzymes, transporters and channel proteins, adhesion proteins, hemoglobins, cellular defense proteins, proteins of the ubiquitin-proteasome system, G-proteins of the Ras family, kinases, chaperone proteins, proteases, translation initiation factors, and others. In addition to the known proteins, there were 43 proteins whose identification was not determined.     

Nitrophorins and related antihemostatic lipocalins from Rhodnius prolixus and other blood-sucking arthropods

Recent gene sequence and crystal structure determinations of salivary proteins from several blood-sucking arthropods have revealed an unusual evolutionary relationship: many such proteins derive their functions from lipocalin protein folds. Many blood-sucking arthropods have independently evolved the ability to overcome a host organism's means of preventing blood loss (called hemostasis). Most blood feeders have proteins that induce vasodilation, inhibit blood coagulation, and reduce inflammation, but do so by distinctly different mechanisms. Despite this diversity, in many cases the antihemostatic activities in such organisms reside in proteins with lipocalin folds. Thirteen such lipocalins are described in this review, with a particular focus on the heme-containing nitrophorins from Rhodnius prolixus, which transport nitric oxide, sequester histamine, and disrupt blood coagulation. Also described are the antiplatelet compounds RPAI, moubatin, and pallidipin from R. prolixus, Ornithodoros moubata, and Triatoma pallidipennis; the antithrombin protein triabin from T. pallidipennis; and the tick histamine binding proteins from Rhipicephalus appendiculatus.     

Proteomic identification of rainbow trout blood plasma proteins and their relationship to seminal plasma proteins

The characterisation of fish blood proteomes is important for comparative studies of seminal and blood proteins as well as for the analysis of fish immune mechanisms and pathways. In this study, LC‐MS/MS and 2D‐DIGE were applied to compare rainbow trout seminal (SP) and blood plasma (BP) proteomes. The 54 differentially abundant proteins identified in SP are involved in a variety of signalling pathways, including protein ubiquitination, liver X receptor/retinoid X receptor (LXR/RXR) and farnesoid X receptor activation, cell cycle and acute phase signalling. These findings may indicate the prevalence of acute phase signalling pathways in trout SP, and its essential role in protecting spermatozoa and reproductive tissues. Our study provides the first in‐depth analysis of the trout BP proteome, with a total of 119 proteins identified. The major proteins of rainbow trout BP were recognised as acute phase proteins. Analysis of BP proteins indicated that acute phase response signalling, the complement system, liver X receptor/retinoid X receptor and farnesoid X receptor activation and the coagulation system are the top canonical pathways. This study enhances knowledge of the blood origin of trout SP proteins and understanding of fish reproductive biology. Our results provide new insight into blood proteins specifically important for fish physiology and innate immunity. The mass spectrometry data are available via ProteomeXchange with the identifier PXD005988 and https://doi.org/10.6019/PXD005988 .     

Water-soluble proteins of Pacinian corpuscles

The composition of water soluble proteins has been investigated in different parts of Pacinian corpuscles. The Pacinian corpuscles fluid proteins have been compared with the receptor homogenate proteins and blood serum of a cat. It is supposed that proteins of the Pacinian corpuscle fluid are produced from the blood system on the whole. The composition of the Pacinian corpuscle fluid as well as a great variety of glycoproteins in it allows to suppose that the Pacinian corpuscles fluid perform a transport function.     

Synthesis of blood and cuticular proteins in late pharate adults of the cecropia silkmoth

Incorporation of tritiated leucine into blood and cuticular proteins of male Hyalophora cecropia was monitored during the final third of pharate adult development. We found no changes in specific activity of the total blood proteins, but there were conspicuous alterations in the banding pattern obtained by acrylamide gel electrophoresis. The specific activity of the cuticular protein extract decreased with age, but the cause of this decline is not obvious. These final stages of exocuticle formation do not involve the appearance or synthesis of new blood or cuticular proteins.     

Identification of Multiple Novel Protein Biomarkers Shed by Human Serous Ovarian Tumors into the Blood of Immunocompromised Mice and Verified in Patient Sera

The most cancer-specific biomarkers in blood are likely to be proteins shed directly by the tumor rather than less specific inflammatory or other host responses. The use of xenograft mouse models together with in-depth proteome analysis for identification of human proteins in the mouse blood is an under-utilized strategy that can clearly identify proteins shed by the tumor. In the current study, 268 human proteins shed into mouse blood from human OVCAR-3 serous tumors were identified based upon human vs. mouse species differences using a four-dimensional plasma proteome fractionation strategy. A multi-step prioritization and verification strategy was subsequently developed to efficiently select some of the most promising biomarkers from this large number of candidates. A key step was parallel analysis of human proteins detected in the tumor supernatant, because substantially greater sequence coverage for many of the human proteins initially detected in the xenograft mouse plasma confirmed assignments as tumor-derived human proteins. Verification of candidate biomarkers in patient sera was facilitated by in-depth, label-free quantitative comparisons of serum pools from patients with ovarian cancer and benign ovarian tumors. The only proteins that advanced to multiple reaction monitoring (MRM) assay development were those that exhibited increases in ovarian cancer patients compared with benign tumor controls. MRM assays were facilely developed for all 11 novel biomarker candidates selected by this process and analysis of larger pools of patient sera suggested that all 11 proteins are promising candidate biomarkers that should be further evaluated on individual patient blood samples.     

Identification of glioma neovascularization-related proteins by using MALDI-FTMS and nano-LC fractionation to microdissected tumor vessels

The identification of angiogenesis-related proteins is important for the development of new antiangiogenic therapies, and such proteins are potential new biomarkers for gliomas. The aim of this study was to identify proteins that are exclusively present in glioma neovasculature and not in the vasculature of normal brain. We combined advanced proteomics techniques to compare the expression profiles of microdissected blood vessels from glioma with blood vessels of normal control brain samples. We measured the enzymatic generated peptide profiles from these microdissected samples by MALDI-FTMS. Subsequently, the samples were fractionated by nano-LC prior to MALDI-TOF/TOF. This combined approach enabled us to identify four proteins that appeared to be exclusively expressed in the glioma blood vessels. Two of these proteins, fibronectin and colligin 2, were validated on tissue sections using specific antibodies. We found that both proteins are present in active angiogenesis in glioma, other neoplasms, and reactive conditions in which neoangiogenesis takes place. This work proves that gel-free mass spectrometric techniques can be used on relatively small numbers of cells generated by microdissection procedures to successfully identify differentially expressed proteins.     

Characteristics of the distribution of long-acting sulfanilamides in purulent pyelonephritis in rats

Distribution of sulfalen, sulfadimethoxine and sulfamethoxypyridazine in the blood and organs of rats and binding of the drugs to the blood serum proteins of the animals with experimental P. aeruginosa pyelonephritis were studied. It was shown that in rats with P. aeruginosa pyelonephritis the levels of long-acting sulfanilamides in the blood and organs were lower, while the levels of their penetration through the histohematic barriers were higher, which was partially due to the decreased binding of sulfanilamides to blood proteins.