Blood Plasma Proteins Research Articles

Effects of carbon sources and plant protein levels in a biofloc system on growth performance, and the immune and antioxidant status of Nile tilapia (Oreochromis niloticus)

The efficacy of different biofloc treatments (BFTs) to compensate for a reduction in dietary protein level under zero-water exchange systems was studied during a 10 weeks experiment, assessing the effect on water quality, growth, immune and antioxidant status of Nile tilapia (Oreochromis niloticus) fingerlings. Six groups were established and fed the same plant-based feed containing 20 or 30% crude protein: two groups in clear water conditions with no added carbon source, two biofloc groups given a wheat milling by-product (WMB) as additional carbon source and two biofloc groups given rice bran (RB). The results showed that biofloc volume was higher when WMB was used as carbon source. The highest growth performance were obtained with the biofloc system and the higher dietary protein level. Fish fed 20% crude protein and stocked in WMB biofloc significantly outperformed the fish fed 30% crude protein and stocked in clear water. Significant improvements in hematocrit, white blood cells, lymphocytes, plasma proteins, and humoral (immunoglobulin, lysozyme, myeloperoxidase and ACH50) and cellular (phagocytosis activity and respiratory burst) immune parameters were observed in all BFT fish. BFT also increased superoxide dismutase and catalase activities. Moreover, the fish fed 20% dietary protein and reared in both biofloc conditions showed equal or superior levels of the immunological criteria to fish fed 30% protein in clear water conditions. In conclusion, using WMB as carbon source could make up for a reduction in dietary protein levels of 10% and improve growth performance, and the immune and antioxidant status of O. niloticus.

Intrinsically Disordered Regions in Serum Albumin: What Are They For?

Serum albumin is a major plasma protein in mammalian blood. The importance of this protein lies in its roles in both bioregulation and transport phenomena. Serum albumin binds various metal ions and participates in the transport and storage of fatty acids, bilirubin, steroids amino acids, and many other ligands, usually with regions of hydrophobic surface. Although the primary role of serum albumin is to transport various ligand, its versatile binding capacities and high concentration mean that it can assume a number of additional functions. The major goal of this article is to show how intrinsic disorder is encoded in the amino acid sequence of serum albumin, and how intrinsic disorder is related to functions of this important serum protein.

The mechanisms and kinetics of initiation of blood coagulation by the extrinsic tenase complex

The system of hemostasis includes coagulation of blood plasma and formation of platelet aggregate. Plasma clotting is a cascade of proteolytic reactions, triggered by the contact of blood plasma with any tissue except the normal vessel endothelium. During the contact an enzymatic complex is formed of the soluble blood plasma protein, factor VIIa, and a membrane-anchored protein, tissue factor. This complex is called extrinsic tenase; it is the key initiator of blood coagulation. The main substrates of extrinsic tenase are blood plasma factors X and IX. During the reaction they undergo proteolytic cleavage and become active serine proteases, factors Xa and IXa, respectively. Factor Xa in complex with its cofactor factor Va catalyzes formation of the key coagulation enzyme, thrombin, which leads to fibrin polymerization and plasma gelation. Although all of the proteins that participate in this process have been known for a long time, several questions remain unanswered. As an example, what is the role of the reaction surface on which the complex is formed, what is the role of membrane-bound multimeres of factor X (Xa), and in what way does the activation of the factor VII proceed? Here, we review recent theoretical and experimental works focused on the biophysical mechanisms of extrinsic tenase functioning and discuss some of these problems.

Hubungan Genetik antara Domba Wonosobo (Dombos), Domba Ekor Tipis (DET) dan Domba Batur (Dombat) Melalui Analisis Polimorfisme Protein Darah

<p>The objective of this study was to evaluate the genetic relationships among Wonosobo sheep (Dombos), Thin-Tailed sheep (DET) and Batur sheep (Dombat) through blood protein polymorphism analysis by using electrophoresis Polyacrylamid Gel Electrophoresis-Thin Layer Electrophoresis (PAGE-TLE). The blood of the 20 head of Dombos (Wonosobo regency), 20 head of DET (Semarang regency) and 20 head of Dombat (Banjarnegara regency) were used as materials. Six locus of blood plasma protein were observed, then the gene frequency, individual heterosigosity, average<br />heterosigosity and genetic distance among three breed were calculated. The results showed that the locus of pre-albumin (Pa), albumin (Alb), ceruloplasmin (Cp), transferrin (Tf), post-transferrin (P-tf) and amylase-I (Am-I), at DET, Dombos and Dombat were polymorphic (0,380-0,454). In conclusion, the DET and Dombat indicated the closer genetic relationship compared to DET-Dombos and Dombos-Dombat.<br />Key words: sheep, polymorphism, genetic relationship</p>

Hubungan Genetik antara Domba Wonosobo (Dombos), Domba Ekor Tipis (DET) dan Domba Batur (Dombat) Melalui Analisis Polimorfisme Protein Darah

<p>The objective of this study was to evaluate the genetic relationships among Wonosobo sheep (Dombos), Thin-Tailed sheep (DET) and Batur sheep (Dombat) through blood protein polymorphism analysis by using electrophoresis Polyacrylamid Gel Electrophoresis-Thin Layer Electrophoresis (PAGE-TLE). The blood of the 20 head of Dombos (Wonosobo regency), 20 head of DET (Semarang regency) and 20 head of Dombat (Banjarnegara regency) were used as materials. Six locus of blood plasma protein were observed, then the gene frequency, individual heterosigosity, average<br />heterosigosity and genetic distance among three breed were calculated. The results showed that the locus of pre-albumin (Pa), albumin (Alb), ceruloplasmin (Cp), transferrin (Tf), post-transferrin (P-tf) and amylase-I (Am-I), at DET, Dombos and Dombat were polymorphic (0,380-0,454). In conclusion, the DET and Dombat indicated the closer genetic relationship compared to DET-Dombos and Dombos-Dombat.<br />Key words: sheep, polymorphism, genetic relationship</p>

Substitution of fish meal by soybean meal in diets for juvenile marbled spinefoot, Siganus rivulatus

ABSTRACTThe present work was performed to assess whether soybean meal (SBM) could replace fish meal (FM) as a source of dietary protein in diets of the marine herbivore Siganus rivulatus. Five iso-nitrogenous (40% protein) and iso-energetic (14 MJ/Kg) diets were prepared with SBM replacing fish meal at 0%, 25%, 50%, 75%, and 100% dietary protein. A 60-day feeding trial was conducted with 240 juvenile S. rivulatus (initial body mass 1.74 ± 0.03 g). Survival, growth, feed, and protein efficiency and hematological parameters were assessed. Growth correlated negatively with increase in dietary SBM. All production parameters measured were negatively affected at all levels of SBM inclusion. There were no effects on total or differential blood counts, hemoglobin, or plasma protein, but hematocrit values were negatively correlated with SBM addition. Results suggest that total replacement of FM by SBM without adding other supplements is not recommended, even for an herbivorous marine fish such as S. rivulatus.

The effect of water dynamics on conformation changes of albumin in pre-denaturation state: photon correlation spectroscopy and simulation

Water is essential for protein three-dimensional structure, conformational dynamics, and activity. Human serum albumin (HSA) is one of major blood plasma proteins, and its functioning is fundamentally determined by the dynamics of surrounding water. The goal of this study is to link the conformational dynamics of albumin to the thermal motions in water taking place in the physiological temperature range. We report the results of photon correlation spectroscopy and molecular dynamics simulations of HSA in aqueous solution. The experimental data processing produced the temperature dependence of the HSA hydrodynamic radius and its zeta potential. Molecular dynamics reproduced the results of experiments and revealed changes in the secondary structure caused by the destruction of hydrogen bonds in the macromolecule's globule.

Investigation of the photophysical and photochemical behavior of substituted zinc phthalocyanines and their water-soluble quaternized derivatives

New nonperipherally and peripherally tetrasubstituted zinc(II) phthalocyanine complexes containing 1-benzylpiperidin-4-ol groups and their quaternized derivatives are reported in this work. These complexes were characterized successfully and their photophysical and photochemical properties were studied. These properties were determined in dimethylsulfoxide for all studied complexes and in water and water $+$ Triton X-100 solutions for ionic Zn(II) phthalocyanines. All studied zinc(II) phthalocyanine complexes generated high singlet oxygen, especially in dimethylsulfoxide, and these properties make them potential photosensitizers for use in the photodynamic therapy of cancer. The effects of the position and quaternization of the substituents on the photophysical and photochemical properties were also examined. The fluorescence of the tetrasubstituted zinc(II) phthalocyanine complexes is effectively quenched by addition of 1,4-benzoquinone. Furthermore, the water soluble Zn(II) phthalocyanines can bind to bovine serum albumin, which is a blood plasma protein.

Effect of Nitrogen Oxide on the Gums <i>Lamina propria</i> Structure and Function in the Experimental Gastritis, Duodenitis and Pharmacological Correction

In the conditions of experi mental chronic gastritis and duodenitis reproduction, carried out among Wistar line 24 rats, was investigated effect of the nitrogen oxide stable metabolites content in the gums homogenates on a lamina propria structure and function. Results of our research have been shown, that level of nitrogen oxide stable metabolites in the rats’ gums tissues homogenates with gastritis and duodenitis decreased in three times, compared with a same period at the intact animals, which indicated about depletion of a nitrogen oxide depot at the given disease. It was revealed a significant decrease typical for the protein and glycoprotein content in the animals gums tissues at the experimental group, which indicated about the catabolic processes predominance. Amount of hexosamines in the gums was significantly increased, which was correlated with a morphological picture (gums lamina propria impregnation with the blood plasma proteins), testified about a change in the vascular wall permeability and microcirculation disruption. In the animals from experimental group was reduced a height of gums papillae connective tissue to 27.18±1.86 x 10-6 m against 56.93±2.64 x 10-6 m in the rats from control group. In the gums lamina propria defines phenomenon of the papillary and reticular layer fibrosis. After medicamentous correction, carried out with an antioxidant and the nitrogen oxide donor was observed repairing structure of the gums lamina propria on a background of increasing microvasculature area. It was determined significantly increased protein and glycoprotein content in the gums tissues with increased levels of nitrogen oxide metabolites.

Transgeneration effects of N-stearoylethanolamine in irradiated rats

to explore possible transgeneration effects in the rats offspring of the first generation of parents subject ed to the combined effects of N stearoylethanolamine (NSE) and external exposure.Materials and metods. In the first generation rats of both sexes born to parents who have experienced the combined influence of ionizing radiation at a dose of 2.0 Gy and NSE a daily dose of 50.0 mg/kg, administered before or after exposure indicators of pro and antioxidant systems (the concentration of TBA reactive products, catalase and glu tathione peroxidase activity in plasma) were defined, concentrations of sex hormones testosterone and estradiol and nitrite anion were studied. Irradiation of parents caused a three fold reduction of testosterone in the blood plasma of males progeny, increased activity of catalase in plasma of female offsprings, as well as significantly increased the concentration of protein in the offsprings' blood plasma of both sexes. Introduction of NSE to parents before exposure caused the acti vation of lipid peroxidation in plasma of both sexes offsprings' against the background of a trustworthy decrease in activity of antioxidant enzymes (catalase and glutathione peroxidase), however, prevented a sharp reduction of testosterone content in the blood plasma of males offsprings, conditioned by the influence of radiation on the body of their parents. NSE introduction to parents after exposure caused no significant violations of pro/ antioxidant bal ance in the body of both sexes progeny, but did not eliminate the negative impact of parental exposure to testos terone levels in male offsprings. The transgeneration impact of NSE is manifested by radio sensitizing properties in the first generation offsprings in case of application to parents before irradiation.

Maghemite nanoparticles coated with human serum albumin: combining targeting by the iron-acquisition pathway and potential in photothermal therapies.

Human serum albumin (HSA), the most abundant plasma protein in human blood, is a natural transport vehicle with multiple ligand binding sites. It, therefore, constitutes an attractive candidate for drug delivery. Targeting may occur via the most known interaction of the protein with the neonatal Fc receptor (FcRn). Here, we investigate another HSA delivery path, involving the transferrin receptor, and we elaborate a maghemite-HSA nanohybrid, opening up new opportunities for medical applications. Fluorescence spectrophotometric titration and size-exclusion chromatography were used to substantiate, in cell-free assays, an interaction between HSA and the transferrin receptor R1. This occurs with a dissociation constant, KD of 6.7 nM. This interaction was confirmed in HeLa cell culture where, by confocal microscopy, rhodamine-labeled HSA is shown to be internalized. HSA was then covalently conjugated onto maghemite nanoparticles (NPs) to give a NP-HSA nanohybrid. The therapeutic potential of this hybrid was demonstrated through its heating capacity in magnetic hyperthermia (MH) and near-infrared (NIR) photothermia (PT). In particular, the Specific Absorption Rate (SAR) in the PT Therapy (PTT) mode, using a 808 nm NIR-LASER (1 W cm-2) and at iron concentration as low as 2.5 mM, was found to be very high, equal to 1870 W g-1 with a temperature increment of 9.2 °C. The nanohybrids incubated with HeLa cells were mainly localized at the cell surface. When the PTT mode was applied under the same conditions as in vitro, mortality was higher in HeLa cells than in fibroblasts (non-malignant cells). Cytotoxicity was checked in both cell lines without the PTT mode; the nanohybrids do not seem to affect cell viability. These results make the nanohybrids very promising agents for NIR-PT and for targeting in cancer therapy, since non-malignant cells were not damaged.

A Method for the Parallel and Sequential Generation of Single-Domain Antibodies for the Proteomic Analysis of Human Blood Plasma

A new efficient method for the parallel and sequential stepwise generation of single-domain antibodies to various high-abundance human-plasma proteins has been described. Single-domain antibodies have a number of features that favorably distinguish them from classical antibodies. In particular, they are able to recognize unusual unique conformational epitopes of native target proteins, small in size, and relatively easily produced and modified; have enhanced stability; and rapidly renature after denaturation. As a consequence, the immunosorbents that utilize these antibodies can be reused without any significant loss of activity. The principal novelty and universality of the described method is that it enables the sequential generation of antibodies to a number of high-abundance and yet unknown antigens of a complex protein mixture without the need for purified antigens. The effectiveness of the method is demonstrated by the example of generation of single-domain antibodies to a number of high-abundance proteins of the human blood plasma. The produced antibodies are promising biotechnological tools that can be used to develop prototypes for new diagnostic and therapeutic agents, as well as appropriate immunoaffinity-based methods for removal, enrichment, analysis, and/or targeting of specified proteins and their complexes from (in) the human blood. As we show, the generated single-domain antibodies can be efficiently used in designing new immunosorbents. As a rule, commercially available analogous immunosorbents that utilize classical antibodies remove many major proteins from the blood plasma immediately, while immunosorbents for many individual proteins are difficult to find and rather expensive. Single-domain antibodies generated by our method are unique new materials that allow for the development of more efficient and delicate approaches to pretreatment of plasma and the analysis of various blood plasma biomarkers.

Lithium ascorbate as a protector of human blood biomolecules under ethanol impact

Background: Alcoholism is an acute social problem worldwide for negative impact on health. This problem is closely associated with social stress and drinking traditions. Previously, it was established exhaustion of blood antioxidant in alcoholism patients. It requires development and investigation of substances with protective properties under ethanol impact, on both cells and biomolecules. Aims and Objectives: Lithium salts are widely used in medicine as mood stabilizers for mental pathologies, including alcoholism. In this work, we investigate lithium ascorbate for the protection of human blood plasma lipids and proteins under ethanol impact. Materials and Methods: Well-known antioxidants - carnosine and ascorbic acid - were used as reference drugs. Heparinized venous blood was used as an experimental substrate. The ethanol and tested substances in the form of water (physiological)solutions were added to blood in vitro. Protection effects were measured and estimated as concentrations of carbonyls proteins and products of lipid peroxidation in blood plasma. Results: It was shown similar protective action of lithium ascorbate and carnosine on human plasma biomolecules against damaging action of ethanol. No protective effect revealed for ascorbic acid. Conclusion: Lithium ascorbate and carnosine were proved to possess the protection effects for blood lipids and proteins under ethanol impact. Normothymic and bloodprotective properties are desirable for psychotropic drugs, and thus, lithium salts could be considered as prospective agents in the treatment of addictions and other pathologies associated with oxidative stress.

Multiscale Modelling of Bionano Interface.

We present a framework for coarse-grained modelling of the interface between foreign nanoparticles (NP) and biological fluids and membranes. Our model includes united-atom presentations of membrane lipids and globular proteins in implicit solvent, which are based on all-atom structures of the corresponding molecules and parameterised using experimental data or atomistic simulation results. The NPs are modelled by homogeneous spheres that interact with the beads of biomolecules via a central force that depends on the NP size. The proposed methodology is used to predict the adsorption energies for human blood plasma proteins on NPs of different sizes as well as the preferred orientation of the molecules upon adsorption. Our approach allows one to rank the proteins by their binding affinity to the NP, which can be used for predicting the composition of the NP-protein corona for the corresponding material. We also show how the model can be used for studying NP interaction with a lipid bilayer membrane and thus can provide a mechanistic insight for modelling NP toxicity.

A comparative study on DNA/BSA binding, DNA photocleavage and antioxidant activities of water soluble peripherally and non-peripherally tetra-3-pyridin-3-ylpropoxy-substituted Mn(III), Cu(II) phthalocyanines

In this study, peripherally and non-peripherally tetra-3-pyridin-3-ylpropoxy substituted Mn(III), Cu(II) phthalocyanines and their water soluble derivatives were synthesized and characterized. To determine DNA binding mode and cleavage pathway of water soluble phthalocyanines 2a, 3a, 2b and 3b were used CT-DNA and supercoiled pBR322 plasmid DNA. The DNA binding of water soluble phthalocyanines monitored using electronic absorption titration, competitive ethidium bromide, thermal denaturation that the water soluble phthalocyanines could bind to CT-DNA via intercalation mode in order 3a > 3b > 2a > 2b. The water soluble phthalocyanines cleaved supercoiled pBR322 plasmid DNA via photocleavage mechanism induced by singlet oxygen under irradiation at 600 nm for 3a, 700 nm for 3b and 750 nm for 2a and 2b. In addition, the binding experiment of BSA showed that water soluble phthalocyanines could bind to blood plasma proteins through a static quenching mechanism. Non-peripheral tetra-substituted water soluble Cu(II) phthalocyanine 3b showed high radical scavenging activities than other water soluble phthalocyanines due to metal ion and non-peripheral position. All of these experiments claimed that non-peripheral tetra-substituted water soluble Cu(II) phthalocyanine 3b have anticancer agent potential due to efficiently biological properties.

Biomonitoring Human Albumin Adducts: The Past, the Present, and the Future.

Serum albumin (Alb) is the most abundant protein in blood plasma. Alb reacts with many carcinogens and/or their electrophilic metabolites. Studies conducted over 20 years ago showed that Alb forms adducts with the human carcinogens aflatoxin B1 and benzene, which were successfully used as biomarkers in molecular epidemiology studies designed to address the role of these chemicals in cancer risk. Alb forms adducts with many therapeutic drugs or their reactive metabolites such as β-lactam antibiotics, acetylsalicylic acid, acetaminophen, nonsteroidal anti-inflammatory drugs, chemotherapeutic agents, and antiretroviral therapy drugs. The identification and characterization of the adduct structures formed with Alb have served to understand the generation of reactive metabolites and to predict idiosyncratic drug reactions and toxicities. The reaction of candidate drugs with Alb is now exploited as part of the battery of screening tools to assess the potential toxicities of drugs. The use of gas chromatography-mass spectrometry, liquid chromatography, or liquid chromatography-mass spectrometry (LC-MS) enabled the identification and quantification of multiple types of Alb xenobiotic adducts in animals and humans during the past three decades. In this perspective, we highlight the history of Alb as a target protein for adduction to environmental and dietary genotoxicants, pesticides, and herbicides, common classes of medicinal drugs, and endogenous electrophiles, and the emerging analytical mass spectrometry technologies to identify Alb-toxicant adducts in humans.

интенсивность свободнорадикальных процессов крови крыс при глубокой гипотермии и в ходе самосогревания

Hypothermic conditions of homoisothermic organisms are characterized by the activation of free-radical processes in tissues. The intensity of these processes occurring at hypothermia is less well understood. The essential increase in heart rate, breathing, blood flow velocity, and metabolic processes during warming must stimulate the generation of reactive oxygen species and oxidative modification of biomolecules. We study the levels of peroxidation markers of lipids (by malondialdehyde) and proteins (by carbonyl groups) in blood plasma and erythrocytes as well as the activity of erythrocyte antioxidant enzymes of rats after deep hypothermia (the temperature in the rectum was 20°C) and self-warming dynamics. A maximum warming rate (0.016°С/min) was revealed over the body temperature range of 22–33°С, below and above these temperatures a warming rate was essentially lower. The warming of rats resulted in a total protein content reduction which negatively correlates (r = –0.967; р<0.05) with a middle molecular peptide level. The deep hypothermia decreased the intensity of oxidative modification of lipid and proteins in blood plasma and red blood cell membranes, and the activity of red blood cell superoxide dismutase (SOD). Maximum amount of products of oxidative modification of lipids and proteins in plasma and erythrocytes membranes caused by rats’ self-warming was observed at body temperature of 30–35°С. After a complete rats’ warming the intensity of oxidative modification of lipids and proteins in plasma and erythrocyte membranes decreased. The activity of SOD and catalase of erythrocyte substantially increased when body temperature reached 35°С. The obtained data indicate that during self-warming at the body temperature of 30–35°С the oxidative stress appears in blood which requires the use of antioxidant defense.

Using of purified platelets for investigation of “cell-efector” model in hemostasis system

Gel filtration on a column of Sepharose 4B was tested as method for separating platelet fraction. The results of cytological and electrophoretic analysis showed absence of other blood cells and plasma proteins in the content of the resulting cell fraction. Obtained platelets were proved to keep morphological stability and retain the ability to aggregate in response to ADP – one of the main physiological inductors

Small Surfactant Concentration Differences Influence Adsorption of Human Serum Albumin on Polystyrene Nanoparticles.

Surfactants, even in miniscule amounts, are often used for the synthesis and especially the stabilization of nanomaterials, which is essential for in vivo applications. In this study, we show that the interaction between nanoparticles and proteins strongly depends on the type of stabilizing surfactants and their (small) concentration changes. The reaction between human serum albumin and polystyrene nanoparticles stabilized by an ionic or nonionic surfactant-sodium dodecyl sulfate or Lutensol AT50, respectively-was monitored using isothermal titration calorimetry. It was found that the amount of surfactant molecules on the surface significantly determines the protein binding affinity and adsorption stoichiometry, which is important for all nanomaterials coming into contact with biological components such as blood plasma proteins. Thus after synthesizing nanomaterials for in vivo applications as drug delivery agents, it is crucial to perform a detailed analysis of the obtained surface chemistry that accounts for the presence of minimal amounts of stabilizing agents.

The Seminal fluid proteome of the polyandrous Red junglefowl offers insights into the molecular basis of fertility, reproductive ageing and domestication.

Seminal fluid proteins (SFPs) are emerging as fundamental contributors to sexual selection given their role in post-mating reproductive events, particularly in polyandrous species where the ejaculates of different males compete for fertilisation. SFP identification however remains taxonomically limited and little is known about avian SFPs, despite extensive work on sexual selection in birds. We characterize the SF proteome of the polyandrous Red junglefowl, Gallus gallus, the wild species that gave rise to the domestic chicken. We identify 1,141 SFPs, including proteins involved in immunity and antimicrobial defences, sperm maturation, and fertilisation, revealing a functionally complex SF proteome. This includes a predominant contribution of blood plasma proteins that is conserved with human SF. By comparing the proteome of young and old males with fast or slow sperm velocity in a balanced design, we identify proteins associated with ageing and sperm velocity, and show that old males that retain high sperm velocity have distinct proteome characteristics. SFP comparisons with domestic chickens revealed both qualitative and quantitative differences likely associated with domestication and artificial selection. Collectively, these results shed light onto the functional complexity of avian SF, and provide a platform for molecular studies of fertility, reproductive ageing, and domestication.