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1

Comparative study of plasma proteins including haptoglobin and serum amyloid A in different types of traumatic reticuloperitonitis in cattle  

Microsoft Academic Search

Traumatic reticuloperitonitis (TRP) is a relatively common disorder in adult cattle which could appear in various forms including\\u000a local peritonitis, diffuse peritonitis, acute pericarditis, and chronic peritonitis. The objective of the present study was\\u000a to compare the changes of plasma proteins including haptoglobin (Hp) and serum amyloid A (SAA) as acute phase proteins (APPs)\\u000a in various types of TRP in

M. Ansari-Lari; S. Nazifi; M. Rezaei; J. Asadi-Fardaqi

2008-01-01

2

Serum heme-albumin: an allosteric protein.  

PubMed

Heme scavenging by plasma proteins, including serum albumin (SA), provides protection against free-heme oxidative damage, limits access by pathogens to the heme, and contributes to iron homeostasis by recycling the heme iron. In turn, serum heme-albumin (SA-heme) acquires heme-based ligand-binding and (pseudo-)enzymatic properties. Heme binding to SA and SA-heme reactivity are allosterically and competitively modulated by endogenous and exogenous third components, this being relevant in pharmacotherapy management. PMID:19946891

Ascenzi, Paolo; Fasano, Mauro

2009-12-01

3

Protein Crystal Serum Albumin  

NASA Technical Reports Server (NTRS)

As the most abundant protein in the circulatory system albumin contributes 80% to colloid osmotic blood pressure. Albumin is also chiefly responsible for the maintenance of blood pH. It is located in every tissue and bodily secretion, with extracellular protein comprising 60% of total albumin. Perhaps the most outstanding property of albumin is its ability to bind reversibly to an incredible variety of ligands. It is widely accepted in the pharmaceutical industry that the overall distribution, metabolism, and efficiency of many drugs are rendered ineffective because of their unusually high affinity for this abundant protein. An understanding of the chemistry of the various classes of pharmaceutical interactions with albumin can suggest new approaches to drug therapy and design. Principal Investigator: Dan Carter/New Century Pharmaceuticals

1998-01-01

4

A dietary pattern including nopal, chia seed, soy protein, and oat reduces serum triglycerides and glucose intolerance in patients with metabolic syndrome.  

PubMed

Metabolic syndrome (MetS) is a health problem throughout the world and is associated with cardiovascular disease and diabetes. Thus, the purpose of the present work was to evaluate the effects of a dietary pattern (DP; soy protein, nopal, chia seed, and oat) on the biochemical variables of MetS, the AUC for glucose and insulin, glucose intolerance (GI), the relationship of the presence of certain polymorphisms related to MetS, and the response to the DP. In this randomized trial, the participants consumed their habitual diet but reduced by 500 kcal for 2 wk. They were then assigned to the placebo (P; n = 35) or DP (n = 32) group and consumed the reduced energy diet plus the P or DP beverage (235 kcal) minus the energy provided by these for 2 mo. All participants had decreases in body weight (BW), BMI, and waist circumference during the 2-mo treatment (P < 0.0001); however, only the DP group had decreases in serum TG, C-reactive protein (CRP), and AUC for insulin and GI after a glucose tolerance test. Interestingly, participants in the DP group with MetS and the ABCA1 R230C variant had a greater decrease in BW and an increase in serum adiponectin concentration after 2 mo of dietary treatment than those with the ABCA1 R230R variant. The results from this study suggest that lifestyle interventions involving specific DP for the treatment of MetS could be more effective if local foods and genetic variations of the population are considered. PMID:22090467

Guevara-Cruz, Martha; Tovar, Armando R; Aguilar-Salinas, Carlos A; Medina-Vera, Isabel; Gil-Zenteno, Lidia; Hernández-Viveros, Isaac; López-Romero, Patricia; Ordaz-Nava, Guillermo; Canizales-Quinteros, Samuel; Guillen Pineda, Luz E; Torres, Nimbe

2012-01-01

5

Protein electrophoresis - serum  

MedlinePLUS

... alpha-2, beta, and gamma globulins. In general, alpha and gamma globulin protein levels increase when there is inflammation in ... globulin: 0.1 to 0.3 g/dL Alpha-2 globulin: 0.6 to 1.0 g/dL Beta globulin: 0.7 to 1.2 g/dL Gamma globulin: 0.7 to 1.6 g/dL ...

6

A Survey of Membrane Proteins in Human Serum  

PubMed Central

Serum and membrane proteins are two of the most attractive targets for proteomic analysis. Previous membrane protein studies tend to focus on tissue sample, while membrane protein studies in serum are still limited. In this study, an analysis of membrane proteins in normal human serum was carried out. Nano-liquid chromatography-electrospray ionization mass spectrometry (NanoLC-ESI-MS/MS) and bioinformatics tools were used to identify membrane proteins. Two hundred and seventeen membrane proteins were detected in the human serum, of which 129 membrane proteins have at least one transmembrane domain (TMD). Further characterizations of identified membrane proteins including their subcellular distributions, molecular weights, post translational modifications, transmembrane domains and average of hydrophobicity, were also implemented. Our results showed the potential of membrane proteins in serum for diagnosis and treatment of diseases. PMID:25288886

Dung, Nguyen Tien; Van Chi, Phan

2012-01-01

7

Piezoelectric microcantilever serum protein detector  

NASA Astrophysics Data System (ADS)

The development of a serum protein detector will provide opportunities for better screening of at-risk cancer patients, tighter surveillance of disease recurrence and better monitoring of treatment. An integrated system that can process clinical samples for a number of different types of biomarkers would be a useful tool in the early detection of cancer. Also, screening biomarkers such as antibodies in serum would provide clinicians with information regarding the patient's response to treatment. Therefore, the goal of this study is to develop a sensor which can be used for rapid, all-electrical, real-time, label-fee, in-situ, specific quantification of cancer markers, e.g., human epidermal receptor 2 (Her2) or antibodies, in serum. To achieve this end, piezoelectric microcantilever sensors (PEMS) were constructed using an 8 mum thick lead magnesium niobate-lead titanate (PMN-PT) freestanding film as the piezoelectric layer. The desired limit of detection is on the order of pg/mL. In order to achieve this goal the higher frequency lateral extension modes were used. Also, as the driving and sensing of the PEMS is electrical, the PEMS must be insulated in a manner that allows it to function in aqueous solutions. The insulation layer must also be compatible with standardized bioconjugation techniques. Finally, detection of both cancer antigens and antibodies in serum was carried out, and the results were compared to a standard commercialized protocol. PEMS have demonstrated the capability of detecting Her2 at a concentration of 5 pg/mL in diluted human serum (1:40) in less than 1 hour. The approach can be easily translated into the clinical setting because the sensitivity is more than sufficient for monitoring prognosis of breast cancer patients. In addition to Her2 detection, antibodies in serum were assayed in order to demonstrate the feasibility of monitoring the immune response for antibody-dependent cellular cytotoxicity (ADCC) in patients on antibody therapies such as Herceptin and Cetuximab. The PEMS displayed a limit of detection of 100 fg/mL, which was 100 times lower than the current methods of protein detection in serum, such as ELISA. Furthermore, the sensitivity of the PEMS device allows it to be capable of determining the dissociation constant, K d, of selective receptors such as antibodies. Using the dose response trials of Her2, Kd has been deduced for H3 scFv, and Herceptin, a commercial antibody specific for Her2.

Capobianco, Joseph A.

8

Distribution of the Serum Proteins of Syrian Hamster as revealed by Starch-gel Electrophoresis  

Microsoft Academic Search

ALTHOUGH considerable work has been published on the distribution of the serum proteins of various laboratory animals, little has been reported on the electrophoretic pattern of the serum proteins of hamster. Moore1 investigated the serum proteins of several species of animals, including that of the hamster, by moving boundary electrophoresis. He identified six components in the serum of hamster corresponding

Abolghassem Amin; K. D. Shamloo

1963-01-01

9

Serum Protein Profile Alterations in Hemodialysis Patients  

SciTech Connect

Background: Serum protein profiling patterns can reflect the pathological state of a patient and therefore may be useful for clinical diagnostics. Here, we present results from a pilot study of proteomic expression patterns in hemodialysis patients designed to evaluate the range of serum proteomic alterations in this population. Methods: Surface-Enhanced Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (SELDI-TOFMS) was used to analyze serum obtained from patients on periodic hemodialysis treatment and healthy controls. Serum samples from patients and controls were first fractionated into six eluants on a strong anion exchange column, followed by application to four array chemistries representing cation exchange, anion exchange, metal affinity and hydrophobic surfaces. A total of 144 SELDI-TOF-MS spectra were obtained from each serum sample. Results: The overall profiles of the patient and control samples were consistent and reproducible. However, 30 well-defined protein differences were observed; 15 proteins were elevated and 15 were decreased in patients compared to controls. Serum from one patient exhibited novel protein peaks suggesting possible additional changes due to a secondary disease process. Conclusion: SELDI-TOF-MS demonstrated dramatic serum protein profile differences between patients and controls. Similarity in protein profiles among dialysis patients suggests that patient physiological responses to end-stage renal disease and/or dialysis therapy have a major effect on serum protein profiles.

Murphy, G A; Davies, R W; Choi, M W; Perkins, J; Turteltaub, K W; McCutchen-Maloney, S L; Langlois, R G; Curzi, M P; Trebes, J E; Fitch, J P; Dalmasso, E A; Colston, B W; Ying, Y; Chromy, B A

2003-11-18

10

Serum protein-binding characteristics of vancomycin.  

PubMed Central

A synthesis of studies of serum protein binding of vancomycin and its reported abnormal binding in serum with very high concentrations of immunoglobulin A (IgA) suggests that this antibiotic may be bound to more than one serum protein. Using an ultrafiltration method for separating free from bound drug and high-performance liquid chromatography to measure drug concentration, we studied the binding characteristics of vancomycin for alpha-1 acid glycoprotein, IgG, IgM, IgA, and albumin. The results showed that vancomycin does not bind to alpha-1 acid glycoprotein, IgG, or IgM. Major binding to albumin and IgA occurs, and total drug binding to serum proteins can be fully explained by binding to these two proteins. We calculated an N (number of binding sites per molecule) of 1.3 +/- 0.4 and a K (association constant) of 3.3 x 10(5) +/- 6.3 x 10(4) M-1 (NK = 4.3 x 10(5) M-1) for binding to IgA, whereas the corresponding NK value for albumin was only 527.5 M-1, indicating that vancomycin preferentially binds to IgA. Very high concentrations of IgA in serum (i.e., grams per deciliter), such as in patients with IgA myeloma, may result in the paradox of high (total) concentrations of vancomycin in serum that may be clinically ineffective. PMID:8517702

Sun, H; Maderazo, E G; Krusell, A R

1993-01-01

11

Two-Dimensional Electrophoresis of Serum Proteins  

Microsoft Academic Search

A METHOD of zone electrophoresis in starch gels has recently been described by one of us1. The high degree of resolution obtained with this method when applied to serum appears to be due to the use of a supporting medium the pore size of which approaches the molecular dimensions of some of the proteins involved, so that resolution by molecular

O. Smithies; M. D. POULIK

1956-01-01

12

The depression of the immune response by serum protein fractions  

PubMed Central

A number of serum protein fractions were tested for their ability to suppress the immune response of rats injected with washed sheep cells. Significant depression of the immune response was produced by human serum proteins, and by bovine ?-globulin fractions. The mechanism of this effect is at present not clear. ImagesFIG. 2 PMID:4162433

Sims, F. H.; Freeman, Jean W.

1966-01-01

13

Interpretation of Serum Calcium in Patients with Abnormal Serum Proteins  

Microsoft Academic Search

Two hundred consecutive specimens received in this laboratory for “liver function tests” showed a wide range of abnormal protein concentrations. Calcium concentration correlated closely with albumin (r = 0·867) but less closely with total protein (r = 0·682). A simple formula for adjusting calcium concentration was derived from the regression equation of calcium on albumin. Adjusted calcium = calcium -

R. B. Payne; A. J. Little; R. B. Williams; J. R. Milner

1973-01-01

14

Serum acute phase proteins and swine health status  

PubMed Central

The purpose of this study was to investigate the relationship between swine health status and the concentration of the serum acute phase proteins, haptoglobin (HP), and C-reactive protein (CRP). A total of 378 clinically healthy pigs from farms A and B, plus 20 pigs culled from farm A due to poor growth, were used in this experiment. Each pig was examined and blood samples were collected during slaughter. The HP concentration was measured by using an HP-hemoglobin binding assay. The CRP concentration was measured by using a CRP enzyme immunoassay. Gross and histopathological lesions were examined and recorded at slaughter. Representative samples were then collected in order to isolate pathogens. Swine enzootic pneumonia, found in 47.7% of the pigs, was the most common lesion. Other lesions included pleuropneumonia (32.7%), suppurative pneumonia (10.3%), fibrinous pericardititis (4.3%), Ascaris migration in the liver (33.9%), and intestinal serositis (3.0%). On farm A, the percentage of pigs with 1 or more lesions was 88.2%. For culled pigs from farm A, the mean serum concentrations of HP and CRP were 2.23 ± 0.14 mg/mL and 252.93 ± 11.62 ?g/mL, which were significantly higher than concentrations in clinically normal pigs (1.42 ± 0.02 mg/mL and 84.88 ± 2.61 ?g/mL, respectively, P < 0.01). Moreover, among clinically normal farm A pigs, the mean HP concentration in pigs with lesions (1.43 ± 0.02 mg/mL) was significantly higher than in pigs without lesions (1.32 ± 0.07 mg/mL) (P < 0.05). However, the mean serum CRP concentrations in these animals were not significantly different. On farm B, the percentage of pigs with one or more lesions was 50.0%. Interestingly, the mean serum HP concentration in clinically normal pigs with lesions was significantly lower in farm B pigs (1.23 ± 0.07 mg/mL) than in the farm A pigs (1.43 ± 0.02 mg/mL; P < 0.01). However, serum CRP concentrations in farm A and B pigs were not significantly different. Serum HP concentration, which is a better indicator of inflammatory reactions in pig herds than serum CRP concentration, provides an important marker for swine health status. PMID:14620865

Chen, Hsin-Hsin; Lin, Jyh-Hung; Fung, Hang-Pong; Ho, Lin-Lin; Yang, Ping-Chin; Lee, Wen-Chuan; Lee, Yan-Pai; Chu, Rea-Min

2003-01-01

15

Human serum and plasma protein binding of enoximone and its sulfoxide metabolite.  

PubMed

Experimental factors and determinants of the protein binding of enoximone (a new cardiotonic agent) were investigated in human serum from healthy, drug-free subjects using a rapid ultrafiltration method; these factors and determinants included nonspecific binding to the apparatus, ultrafiltrate volume, temperature, serum pH, enoximone serum concentration, and enoximone sulfoxide (metabolite) concentration. It was demonstrated from mass balance experiments that nonspecific binding to the apparatus did not occur. Within the range investigated, ultrafiltrate volume did not affect the binding result. However, serum pH and temperature were critical variables. At pH 7.4 and 37 degrees C, enoximone serum binding occurred to the extent of approximately 70%; over the therapeutic serum concentration range, this binding was concentration independent. Experiments with purified albumin solutions indicated that much of the serum binding could be accounted for by albumin. At concentrations exceeding those observed clinically, enoximone sulfoxide did not affect enoximone serum binding. In another experiment, enoximone binding to serum was compared with that from plasma containing either heparin or disodium EDTA. There were essentially no differences. Enoximone sulfoxide serum protein binding was also investigated in serum from healthy, drug-free human subjects; binding occurred to the extent of approximately 5%. PMID:3244103

Hook, R H; Boxenbaum, H; Thompson, G A; Okerholm, R A

1988-12-01

16

BINDING OF PERFLUORINATED FATTY ACIDS TO SERUM PROTEINS  

Microsoft Academic Search

Perfluorooctane sulfonic acid (PFOS) accumulates in the liver and blood of exposed organisms. The potential for these surfactant molecules to interfere with hormone\\/protein interactions in blood is of concern given the importance of these interactions. The PFOS binding to serum proteins was investigated by assessing its ability to displace a variety of steroid hormones from specific binding proteins in the

Paul D. Jones; Wenyue Hu; Wim De Coen; John L. Newsted; John P. Giesy

2003-01-01

17

Interaction of Serum Proteins with Surface of Hemodialysis Fiber Membranes  

NASA Astrophysics Data System (ADS)

The poly(vinyl pyrrolidone)-covered hydrophilic surface of hollow-fiber membranes (fiber membrane, hereafter) for hemodialysis was mechanically probed using modified tips on an atomic force microscope (AFM) with covalent crosslinkers and several types of serum protein. The retraction part of many of the force extension (F-E) curves obtained with AFM tips coated with serum albumin had a long and smooth extension up to 200-300 nm indicating forced elongation of poly(vinyl pyrrolidone) chains. When fibrinogen-coated tips were used, long extension F-E curves up to 500 nm with multiple peaks were obtained in addition to smooth curves most likely reflecting the unfolding of fibrinogen molecules. The results indicated that individual polymer chains had a significant affinity toward serum proteins. The adhesion frequency of tips coated with serum proteins was lower on the poly(vinyl pyrrolidone) surface than on the uncoated hydrophobic polysulfone surface.

Afrin, Rehana; Shirako, Yuji; Kishimoto, Kikuo; Ikai, Atsushi

2012-08-01

18

INFLUENCE OF X-RAY IRRADIATION UPON SERUM PROTEINS  

Microsoft Academic Search

Changes of serum proteins of x irradiatsd rabbits were used, total ; protein and albumin decreased as the total dose increased. In the early period, ; alpha globuin increased and gamma globulin decreased, wbile in the later ; period, alpha globulin decreased and gamma globulin increased. More marked ; changes were observed after liver irradiation than after irradiation of the

Shibata

1960-01-01

19

THE INFLUENCE OF ROENTGEN IRRADIATION UPON SERUM PROTEIN  

Microsoft Academic Search

Serum protein fractions of x irradiated rabbits were studied by paper ; electrophoresis. Total protein, the albumin fraction, and the A\\/G ratio were ; decreased in all irradiated animals, while the alpha -, BETA -, and gamma -; globulin fractions were increased. These changes did not vary with the x-ray ; dose when single irradiations were applied but they did

Shibata

1961-01-01

20

Binding of aminoglycoside antibiotics to human serum proteins  

Microsoft Academic Search

Summary The binding of several aminoglycoside antibiotics to human serum proteins was studied under varying experimental conditions. The protein binding was determined by means of equilibrium dialysis and, with sisomicin and gentamicin, also by the ultracentrifuge method in the presence and in the absence of Ca++ and Mg++ ions. The technical experimental procedure (dialysis, ultrafiltration, ultracentrifugation) has no effect on

H. Rosenkranz; W. Scholtan; M. Scheer

1978-01-01

21

Biologically active protein fragments containing specific binding regions of serum albumin or related proteins  

NASA Technical Reports Server (NTRS)

In accordance with the present invention, biologically active protein fragments can be constructed which contain only those specific portions of the serum albumin family of proteins such as regions known as subdomains IIA and IIIA which are primarily responsible for the binding properties of the serum albumins. The artificial serums that can be prepared from these biologically active protein fragments are advantageous in that they can be produced much more easily than serums containing the whole albumin, yet still retain all or most of the original binding potential of the full albumin proteins. In addition, since the protein fragment serums of the present invention can be made from non-natural sources using conventional recombinant DNA techniques, they are far safer than serums containing natural albumin because they do not carry the potentially harmful viruses and other contaminants that will be found in the natural substances.

Carter, Daniel C. (Inventor)

1998-01-01

22

Erythropoietin binding protein from mammalian serum  

DOEpatents

Purified mammalian erythropoietin binding-protein is disclosed, and its isolation, identification, characterization, purification, and immunoassay are described. The erythropoietin binding protein can be used for regulation of erythropoiesis by regulating levels and half-life of erythropoietin. A diagnostic kit for determination of level of erythropoietin binding protein is also described.

Clemons, Gisela K. (Berkeley, CA)

1997-01-01

23

Magnetic nanoparticles-serum proteins bioconjugates for binding of irinotecan.  

PubMed

The binding of irinotecan to serum proteins (hemoglobin, globulin and human serum albumin) was studied on the surface of epoxide modified superparamagnetic iron oxide nanoparticles (GPTS-SPIONs), which were synthesized by the coprecipitation of ferrous and ferric salts with NH4OH and then modified with [3-(2,3-epoxypropoxy)propyl] trimethoxy silane (GPTS) to obtain functional epoxide groups on the SPIONs' surface. Results were compared to find an alternative as drug carries system. Data showed that binding amount of human serum albumin (HSA), globulin (Glb) and hemoglobin (Hb) found to be as 44, 21.2 and 32.6?g per 20mg of GPTS modified SPIONs, respectively. The thermal behavior of the serum protein-Ir interaction on GPTS-SPIONs was also studied by using thermo gravimetric analysis (TGA) technique and then the kinetic parameters for the thermal decomposition were determined using Horowitz-Metzger method. PMID:25445689

Tamyurek, Ecem; Maltas, Esra; Bas, Salih Zeki; Ozmen, Mustafa; Yildiz, Salih

2015-02-01

24

Serum protein alterations in dogs naturally infected with Toxoplasma gondii  

Microsoft Academic Search

We conducted this study to describe the serum electrophoretic pattern in dogs associated with the infection of Toxoplasma gondii (T. gondii). The serum protein pattern of 25 dogs with confirmed T. gondii infection and 15 clinically healthy dogs were evaluated using native polyacrylamide gel electrophoresis. Albumin, alpha-1\\u000a globulin, alpha-2 globulin, beta globulin, and gamma globulin bands were seen from the

Gul Fatma Yarim; Cevat Nisbet; Taraneh Oncel; Sena Cenesiz; Gulay Ciftci

2007-01-01

25

Identification of Differentially Expressed Serum Proteins in Infectious Purpura Fulminans  

PubMed Central

Purpura fulminans (PF) is a life-threatening hemorrhagic condition. Because of the rarity and randomness of the disease, no improvement in treatment has been made for a long time. In this study, we assessed the serum proteome response to PF by comparing serum proteins between healthy controls and PF patient. Liquid chromatography with tandem mass spectrometry (LC-MS/MS) approach was used after depleting 6 abundant proteins of serum. In total, 262 proteins were confidently identified with 2 unique peptides, and 38 proteins were identified significantly up- (?2) or downregulated (?0.5) based on spectral counting ratios (SpCPF/N). In the 38 proteins with significant abundance changes, 11 proteins were previously known to be associated with burn or sepsis response, but 27 potentially novel proteins may be specifically associated with PF process. Two differentially expressed proteins, alpha-1-antitrypsin (SERPINA1) and alpha-2 antiplasmin (SERPINF2), were validated by Western blot. This is the first study where PF patient and healthy controls are compared in a proteomic study to elucidate proteins involved in the response to PF. This study provides an initial basis for future studies of PF, and the differentially expressed proteins might provide new therapeutic targets to decrease the mortality of PF. PMID:24659849

Hu, Jiong-yu; Han, Jian; Zhang, Dong-xia; Jiang, Xu-pin; Chen, Bing; Huang, Yue-sheng

2014-01-01

26

Immunoglobulin G antibodies directed against protein III block killing of serum-resistant Neisseria gonorrhoeae by immune serum  

PubMed Central

Neisseria gonorrhoeae that resist complement-dependent killing by normal human serum (NHS) are sometimes killed by immune convalescent serum from patients recovering from disseminated gonococcal infection (DGI). In these studies, killing by immune serum was prevented or blocked by IgG isolated from NHS. Purified human IgG antibodies directed against gonococcal protein III, an antigenically conserved outer membrane protein, contained most of the blocking activity in IgG. Antibodies specific for gonococcal porin (protein I), the major outer membrane protein, displayed no blocking function. In separate experiments, immune convalescent DGI serum which did not exhibit bactericidal activity was restored to killing by selective depletion of protein III antibodies by immunoabsorption. These studies indicate that protein III antibodies in normal and immune human serum play a role in serum resistance of N. gonorrhoeae. PMID:3095479

1986-01-01

27

Distribution of Serum Total Protein in Elderly Chinese  

PubMed Central

The serum total protein levels of the elderly possibly decrease gradually with aging. However, serum total protein levels are not suitable as a uniform reference standard for the elderly at different ages and genders. Thus, we investigated the total serum protein distribution in different gender and age groups of 11,453 elderly individuals aged ?60 years and without liver or renal disease from Lianyungang, Jiangsu, China. The total protein levels (TPL) of these individuals exhibited normal distribution (Z?=?1.206, P?=?0.109), whereas the reference range (95% CI) was 54.1 g/L to 82.3 g/L. TPL was higher in females than in males for those aged between 60 and 75 years, whereas no significant difference was observed for those aged between 80 and 95 years. TPL was negatively correlated with age in males (r?=??0.1342, P<0.05), females (r?=??0.304, P<0.05), and the total group (r?=??0.2136, P<0.05). TPL also decreased with aging and showed a faster rate in women than in men. These results indicated that an appropriate range of serum total protein based on age and gender differences should be used for clinical applications. PMID:24967900

Tian, Chang-Rong; Qian, Li; Shen, Xiao-Zhu; Li, Jia-Jing; Wen, Jiang-Tao

2014-01-01

28

Serum and saliva protein levels in females with breast cancer  

PubMed Central

The aim of the present study was to investigate the change in the total protein content between the serum and saliva of female patients with breast cancer and in healthy females. The study was conducted between October 2012 and November 2013. There were 80 females in the present study with 40 breast cancer patients and 40 healthy control subjects, with an age range of 50–70 years. The results of the study showed that the mean value ± standard deviation of the total serum protein in patients with breast cancer was 7.63±0.41 g/dl, whereas in the healthy subjects it was 6.14±1.84 g/dl. The total salivary protein measurement was 0.14±0.07 g/dl and 0.25±0.09 g/dl in the breast cancer and healthy group, respectively. Therefore, it can be concluded that the total serum protein was higher in female patients with breast cancer, whereas the levels in the saliva were lower compared to the healthy female group. The results of the present study indicate that serum protein levels may be used for the diagnosis of breast cancer. PMID:25364460

AL-MUHTASEB, SABAH ISA

2014-01-01

29

Serum and saliva protein levels in females with breast cancer.  

PubMed

The aim of the present study was to investigate the change in the total protein content between the serum and saliva of female patients with breast cancer and in healthy females. The study was conducted between October 2012 and November 2013. There were 80 females in the present study with 40 breast cancer patients and 40 healthy control subjects, with an age range of 50-70 years. The results of the study showed that the mean value ± standard deviation of the total serum protein in patients with breast cancer was 7.63±0.41 g/dl, whereas in the healthy subjects it was 6.14±1.84 g/dl. The total salivary protein measurement was 0.14±0.07 g/dl and 0.25±0.09 g/dl in the breast cancer and healthy group, respectively. Therefore, it can be concluded that the total serum protein was higher in female patients with breast cancer, whereas the levels in the saliva were lower compared to the healthy female group. The results of the present study indicate that serum protein levels may be used for the diagnosis of breast cancer. PMID:25364460

Al-Muhtaseb, Sabah Isa

2014-12-01

30

Serum Protein Signatures Differentiating Autoimmune Pancreatitis versus Pancreatic Cancer  

PubMed Central

Autoimmune pancreatitis (AIP) is defined by characteristic lymphoplasmacytic infiltrate, ductal strictures and a pancreatic enlargement or mass that can mimic pancreatic cancer (PaCa). The distinction between this benign disease and pancreatic cancer can be challenging. However, an accurate diagnosis may pre-empt the misdiagnosis of cancer, allowing the appropriate medical treatment of AIP and, consequently, decreasing the number of unnecessary pancreatic resections. Mass spectrometry (MS) and two-dimensional differential gel electrophoresis (2D-DIGE) have been applied to analyse serum protein alterations associated with AIP and PaCa, and to identify protein signatures indicative of the diseases. Patients' sera were immunodepleted from the 20 most prominent serum proteins prior to further 2D-DIGE and image analysis. The identity of the most-discriminatory proteins detected, was performed by MS and ELISAs were applied to confirm their expression. Serum profiling data analysis with 2D-DIGE revealed 39 protein peaks able to discriminate between AIP and PaCa. Proteins were purified and further analysed by MALDI-TOF-MS. Peptide mass fingerprinting led to identification of eleven proteins. Among them apolipoprotein A-I, apolipoprotein A-II, transthyretin, and tetranectin were identified and found as 3.0-, 3.5-, 2-, and 1.6-fold decreased in PaCa sera, respectively, whereas haptoglobin and apolipoprotein E were found to be 3.8- and 1.6-fold elevated in PaCa sera. With the exception of haptoglobin the ELISA results of the identified proteins confirmed the 2D-DIGE image analysis characteristics. Integration of the identified serum proteins as AIP markers may have considerable potential to provide additional information for the diagnosis of AIP to choose the appropriate treatment. PMID:24349355

Fritz, Stefan; Hinz, Ulf; Schnölzer, Martina; Kempf, Tore; Warnken, Uwe; Michel, Angelika; Pawlita, Michael; Werner, Jens

2013-01-01

31

Targeted quantification of low ng/mL level proteins in human serum without immunoaffinity depletion  

PubMed Central

We recently reported an antibody-free targeted protein quantification strategy, termed high-pressure, high-resolution separations with intelligent selection and multiplexing (PRISM) for achieving significantly enhanced sensitivity using selected reaction monitoring (SRM) mass spectrometry. Integrating PRISM with front-end IgY14 immunoaffinity depletion, sensitive detection of targeted proteins at 50–100 pg/mL levels in human blood plasma/serum was demonstrated. However, immunoaffinity depletion is often associated with undesired losses of target proteins of interest. Herein we report further evaluation of PRISM-SRM quantification of low-abundance serum proteins without immunoaffinity depletion. Limits of quantification (LOQ) at low ng/mL levels with a median coefficient of variation (CV) of ~12% were achieved for proteins spiked into human female serum. PRISM-SRM provided >100-fold improvement in the LOQ when compared to conventional LC-SRM measurements. PRISM-SRM was then applied to measure several low-abundance endogenous serum proteins, including prostate-specific antigen (PSA), in clinical prostate cancer patient sera. PRISM-SRM enabled confident detection of all target endogenous serum proteins except the low pg/mL-level cardiac troponin T. A correlation coefficient >0.99 was observed for PSA between the results from PRISM-SRM and immunoassays. Our results demonstrate that PRISM-SRM can successful quantify low ng/mL proteins in human plasma or serum without depletion. We anticipate broad applications for PRISM-SRM quantification of low-abundance proteins in candidate biomarker verification and systems biology studies. PMID:23763644

Shi, Tujin; Sun, Xuefei; Gao, Yuqian; Fillmore, Thomas L.; Schepmoes, Athena A.; Zhao, Rui; He, Jintang; Moore, Ronald J.; Kagan, Jacob; Rodland, Karin D.; Liu, Tao; Liu, Alvin Y.; Smith, Richard D.; Tang, Keqi; Camp, David G.; Qian, Wei-Jun

2013-01-01

32

Amyloid-related serum component (protein ASC) IN LEPROSY PATIENTS.  

PubMed Central

The presence of amyloid-related serum component, protein ASC, in serum samples from 63 leprosy patients was investigated. Protein ASC was detected in 38% of the patients. A correlation to the disease spectrum of leprosy was apparent: polar lepromatous cases, 64% positive; borderline lepromatous, 50%; borderline tuberculoid, 36%; subpolar tuberculoid, 17%; and polar tuberculoid, negative. Antibody activity against the a antigen of Mycobacterium leprae was also determined, showing a similar correlation to the disease spectrum. Serum samples from 23 apparently healthy Ethiopians serving as controls showed a protein ASC incidence of 22%. This figure is significantly higher than the frequency found by others among healthy Norwegian blood donors. Immunoglobulin M levels among patients were elevated in the borderline lepromatous and poplar lepromatous groups. The three tuberculoid groups did not differ in this respect from the control group but were all elevated as compared to a normal Caucasian serum pool. Although raised immunoglobulin M levels seemed to parallel increased frequencies of protein ASC in the patient groups as well as in controls, this correlation might be only secondary to a primary derangement in T-cell function. PMID:804451

Kronvall, G; Husby, G; Samuel, D; Bjune, G; Wheate, H

1975-01-01

33

Distribution of alkaline phosphatase in serum protein fractions  

Microsoft Academic Search

A technique for measuring the alkaline phosphatase activity of serum protein fractions separated by electrophoresis on cellulose acetate strips is described.Alkaline phosphatase activity in alpha 2, alpha 1, and beta globulins and in albumin is present in varying proportions in normal and pathological sera. In normal sera peak activity is in the alpha 2 globulins and beta globulins have a

N. H. Korner

1962-01-01

34

CHANGES IN RABBIT SERUM PROTEIN FOLLOWING X-RAY IRRADIATION  

Microsoft Academic Search

The effect of radiation on serum protein fractions was rather unstable ; after x irradiation with doses smaller than 200 r; however, when doses of x rays ; up to 1000 r were givcn, deiinite changes such as increase of alpha -globulin ; and decrease of gamma -globulin resulted. (Abstr. Japan Med., li No. l3, l96l);

Arai

1960-01-01

35

Serum protein changes during exposure to ozone. [New Zealand Rabbits  

Microsoft Academic Search

Serial measurements of serum proteins in rabbits exposed to 0.4 ppM and 1.0 ppM of ozone revealed progressive changes in the concentrations of the various fractions: albumin concentration declined while ..cap alpha.. and ..gamma..-globulin concentrations increased.

A. Y. S. Pan; Z. Jegier

1976-01-01

36

Lyophilic Properties of Isolated Serum-Protein Fractions  

Microsoft Academic Search

ELECTROPHORESIS on paper of serum proteins has enabled us to separate small quantities into albumin, and alpha-, beta- and gamma-globulin. By using a comparatively thick filter paper (MunktellNr. 20, Grycksbo, Sweden), 58 cm. × 7 cm., we can separate up to 30 mgm. of protein within 24 hr., with 5.5 V.\\/cm. and 16-18 m.amp. applied in a closed atmosphere. As

Ch. Wunderly

1952-01-01

37

Serum proteins of wild turkey vultures (Cathartes aura).  

PubMed

We separated and identified the major serum proteins of turkey vultures (Cathartes aura): albumin, HDL, LDL, IgG and M, transferrin, alpha 2M and putatively, IgA, ceruloplasmin and fibronectin. Turkey vulture HDL and LDL had similar electrophorectic mobility and solution properties as those from gallinaceous birds. Turkey vulture IgG and M, and their subunits, had molecular weights comparable to other birds. Serum IgG and M levels in wild turkey vultures were within the range of values reported for domestic avian species. PMID:6661906

Apanius, V; Temple, S A; Bale, M

1983-01-01

38

Nongradient blue native gel analysis of serum proteins and in-gel detection of serum esterase activities.  

PubMed

The objective of the present study was to analyze serum protein complexes and detect serum esterase activities using nongradient blue native polyacrylamide gel electrophoresis (BN-PAGE). For analysis of potential protein complexes, serum from rat was used. Results demonstrate that a total of 8 gel bands could be clearly distinguished after Coomassie blue staining, and serum albumin could be isolated nearly as a pure protein. Moreover, proteins in these bands were identified by electrospray mass spectrometry and low-energy collision induced dissociation (CID)-MS/MS peptide sequencing and the existence of serum dihydrolipoamide dehydrogenase (DLDH) was confirmed. For studies of in-gel detection of esterase activities, serum from rat, mouse, and human was used. In-gel staining of esterase activity was achieved by the use of either ?-naphthylacetate or ?-naphthylacetate in the presence of Fast blue BB salt. There were three bands exhibiting esterase activities in the serum of both rat and mouse. In contrast, there was only one band showing esterase activity staining in the human serum. When serum samples were treated with varying concentrations of urea, esterase activity staining was abolished for all the bands except the one containing esterase 1 (Es1) protein that is known to be a single polypeptide enzyme, indicating that majority of these esterases were protein complexes or multimeric proteins. We also identified the human serum esterase as butyrylcholinesterase following isolation and partial purification using ammonium sulfate fractioning and ion exchange column chromatographies. Where applicable, demonstrations of the gel-based method for measuring serum esterase activities under physiological or pathophysiological conditions were illustrated. Results of the present study demonstrate that nongradient BN-PAGE can serve as a feasible analytical tool for proteomic and enzymatic analysis of serum proteins. PMID:21237726

Thangthaeng, Nopporn; Sumien, Nathalie; Forster, Michael J; Shah, Ruchir A; Yan, Liang-Jun

2011-02-15

39

Serum immune-related proteins are differentially expressed during hibernation in the American black bear.  

PubMed

Hibernation is an adaptation to conserve energy in the face of extreme environmental conditions and low food availability that has risen in several animal phyla. This phenomenon is characterized by reduced metabolic rate (?25% of the active basal metabolic rate in hibernating bears) and energy demand, while other physiological adjustments are far from clear. The profiling of the serum proteome of the American black bear (Ursus americanus) may reveal specific proteins that are differentially modulated by hibernation, and provide insight into the remarkable physiological adaptations that characterize ursid hibernation. In this study, we used differential gel electrophoresis (DIGE) analysis, liquid chromatography coupled to tandem mass spectrometry, and subsequent MASCOT analysis of the mass spectra to identify candidate proteins that are differentially expressed during hibernation in captive black bears. Seventy serum proteins were identified as changing by ±1.5 fold or more, out of which 34 proteins increased expression during hibernation. The majority of identified proteins are involved in immune system processes. These included ?2-macroglobulin, complement components C1s and C4, immunoglobulin ? and J chains, clusterin, haptoglobin, C4b binding protein, kininogen 1, ?2-HS-glycoprotein, and apoplipoproteins A-I and A-IV. Differential expression of a subset of these proteins identified by proteomic analysis was also confirmed by immunodetection. We propose that the observed serum protein changes contribute to the maintenance of the hibernation phenotype and health, including increased capacities for bone maintenance and wound healing during hibernation in bears. PMID:23825529

Chow, Brian A; Donahue, Seth W; Vaughan, Michael R; McConkey, Brendan; Vijayan, Mathilakath M

2013-01-01

40

Rheology of globular proteins: apparent yield stress, high shear rate viscosity and interfacial viscoelasticity of bovine serum albumin solutions  

E-print Network

biological functions, including maintaining blood pH and osmotic pressure,2 as well as transporting ligands circulation.1 The concentration of human serum albumin (HSA) in blood plasma is $40 mg mlÃ?1 ($0.6 mM). Bovine are the most abundant among the constituent proteins in mammalian blood.1 Serum albumins participate in various

41

CHANGES IN SERUM PROTEINS OF RATS SUBJECTED TO SMALL DOSES OF X RAYS. I. TOTAL PROTEINS  

Microsoft Academic Search

Serum protein levels were followed by paper electrophoresis in rats ; exposed to 180-kv x rays either over the whole body or in the liver region. The ; doses in each instance were 50 to 80 r, and in some rats the same total doses ; were given as four daily exposures on consecutive days. The serums were examined ;

F. A. Paolini; A. Pannella

1961-01-01

42

Pathology Case Study: Monoclonal Protein in Serum  

NSDL National Science Digital Library

This is a case study presented by the University of Pittsburgh Department of Pathology in which a 47-year-old man working in the paint industry with a complicated past medical history is hospitalized and treated over the course of a year. Visitors are given a summary of all of the patient's visits and test results, including images. A final diagnosis is given, with notes by the attending doctors, along with references. This is an excellent resource for students in the health sciences to familiarize themselves with using patient history and laboratory results to diagnose disease. It is also a helpful site for educators to use to introduce or test student learning in clinical immunology.

Kelly, Robert

43

Serum protein fractions in rheumatoid pneumoconiosis without arthritis  

Microsoft Academic Search

Fractionation of the serum proteins by filter-paper electrophoresis in 14 coal-miners who had the characteristic radiological opacities of rheumatoid pneumoconiosis but no evidence of rheumatoid arthritis showed a reduction in the mean level of albumin and increases in the alpha-2 and gamma-globulins compared with the values in non-arthritic miners with simple coal-workers' pneumoconiosis and in normal subjects. The changes were

R. B. Payne

1962-01-01

44

Electrophoretic Studies of Serum Proteins of Foetal Rabbits  

Microsoft Academic Search

The sera of non-pregnant adult rabbits which had been hyperimmunized to Brucella abortus antigen, the sera of pregnant female rabbits which had not been immunized, and the sera, exocoelomic fluids, amniotic fluids and stomach contents of 25-day-old foetal rabbits were examined electrophoretically and ultracentrifugally. The serum of pregnant rabbits differed both in total protein concentration and in the proportions of

F. W. R. Brambell; W. A. Hemmings; M. Henderson; R. A. Kekwick

1953-01-01

45

Serum-derived bovine immunoglobulin/protein isolate: postulated mechanism of action for management of enteropathy  

PubMed Central

The health and performance of the gastrointestinal tract is influenced by the interaction of a variety of factors, including diet, nutritional status, genetics, environment, stress, the intestinal microbiota, immune status, and gut barrier. Disruptions in one or more of these factors can lead to enteropathy or intestinal disorders that are known to occur in concert with certain disease states or conditions such as irritable bowel syndrome or human immunodeficiency virus (HIV) infection. Nutritional support in the form of a medical food along with current therapies could help manage the adverse effects of enteropathy, which include effects on nutrient digestion, absorption, and metabolism, as well as utilization of nutrients from foodstuffs. Numerous studies have demonstrated that oral administration of plasma- or serum-derived protein concentrates containing high levels of immunoglobulins can improve weight management, normalize gut barrier function, and reduce the severity of enteropathy in animals. Recent trials in humans provide preliminary evidence that a serum-derived bovine immunoglobulin/protein isolate is safe and improves symptoms, nutritional status, and various biomarkers associated with enteropathy in patients with HIV infection or diarrhea-predominant irritable bowel syndrome. This review summarizes data from preclinical and clinical studies with immunoglobulin-containing plasma/serum protein concentrates, with a focus on the postulated mode of action of serum-derived bovine immunoglobulin/protein isolate for patients with enteropathy. PMID:24904221

Petschow, Bryon W; Burnett, Bruce; Shaw, Audrey L; Weaver, Eric M; Klein, Gerald L

2014-01-01

46

Homology of Serum Proteins of Golden Shiner (Notemigonus crysoleucas) and Man  

Microsoft Academic Search

Comparing properties of electrophoretically-separated proteins of the golden shiner with those of man permitted an evaluation of the suitability of traditional terminology for naming the proteins of fishes. Collectively, electrophoretic mobility, percentage composition, solubility properties, and distribution of lipoproteins of six golden shiner serum proteins differed sufficiently from the classical definitions of serum proteins to prevent homologizing the proteins of

Robert C. Summerfelt

1966-01-01

47

Tuning the serum persistence of human serum albumin domain III:diabody fusion proteins  

PubMed Central

The long circulation persistence of human serum albumin (HSA) is enabled by its domain III (DIII) interaction with the neonatal Fc receptor (FcRn). A protein scaffold based on HSA DIII was designed. To modify the serum half life of the scaffold, residues H535, H510, and H464 were individually mutated to alanine. HSA DIII wild type (WT) and variants were fused to the anti-carcinoembryonic antigen (CEA) T84.66 diabody (Db), radiolabeled with 124I and injected into xenografted athymic mice for serial PET/CT imaging. All proteins targeted the CEA-positive tumor. The mean residence times (MRT) of the proteins, calculated by quantifying blood activity from the PET images, were: Db-DIII WT (56.7 h), H535A (25 h), H510A (20 h), H464A (17 h), compared with Db (2.9 h). Biodistribution confirmed the order of blood clearance from slow to fast: Db-DIII WT > H535A > H510A > H464A > Db with 4.0, 2.0, 1.8, 1.6 and 0.08 %ID/g of remaining blood activity at 51 h, respectively. This study demonstrates that attenuating the DIII–FcRn interaction provides a way of controlling the pharmacokinetics of the entire Db-DIII fusion protein without compromising tumor targeting. H464 appears to be most crucial for FcRn binding (greatest reduction in MRT), followed by H510 and H535. By mutating the DIII scaffold, we can dial serum kinetics for imaging or therapy applications. PMID:20802234

Kenanova, Vania E.; Olafsen, Tove; Salazar, Felix B.; Williams, Lawrence E.; Knowles, Scott; Wu, Anna M.

2010-01-01

48

Interactions of apomorphine with serum and tissue proteins  

SciTech Connect

Physical and covalent interactions of apomorphine with serum and tissue proteins could influence the drug's disposition and pharmacological activities in mammals. Ultrafiltration, equilibrium dialysis, and ultraviolet spectrophotometric methods have been used to study the reversible binding of apomorphine to bovine, human, rat, and swine plasma proteins. The degree of binding was generally greater than 90%, but variations were noted in some instances on the basis of drug concentrations and pH over the range of 6.8-7.8. Incubation of (8,9-/sup 3/H2)apomorphine with bovine serum albumin led to retention of radioactivity and a stoichiometrically controlled released of tritium which arose from the reaction of an electrophilic drug oxidation product and protein, producing drug-protein conjugates. In vitro experiments with mouse striatal brain preparations indicated parallel covalent binding reactions. In vivo experiments in mice indicated accumulation of radioactivity in brain regions and other tissues following daily injections of (8,9-/sup 3/H2)apomorphine for 14 days. The physical and covalent interactions of apomorphine with mammalian tissue proteins could be the cause of longer disposition half-lives in mammals than those previously reported. The covalent interactions, in particular, may be important in elucidating the mechanism of apomorphine-induced behavioral effects in mice.

Smith, R.V.; Velagapudi, R.B.; McLean, A.M.; Wilcox, R.E.

1985-05-01

49

Unraveling the mysteries of serum albumin—more than just a serum protein  

PubMed Central

Serum albumin is a multi-functional protein that is able to bind and transport numerous endogenous and exogenous compounds. The development of albumin drug carriers is gaining increasing importance in the targeted delivery of cancer therapy, particularly as a result of the market approval of the paclitaxel-loaded albumin nanoparticle, Abraxane®. Considering this, there is renewed interest in isolating and characterizing albumin-binding proteins or receptors on the plasma membrane that are responsible for albumin uptake. Initially, the cellular uptake and intracellular localization of albumin was unknown due to the large confinement of the protein within the vascular and interstitial compartment of the body. Studies have since assessed the intracellular localization of albumin in order to understand the mechanisms and pathways responsible for its uptake, distribution and catabolism in multiple tissues, and this is reviewed herein. PMID:25161624

Merlot, Angelica M.; Kalinowski, Danuta S.; Richardson, Des R.

2014-01-01

50

Usual Intake of Total protein foods including beans and peas  

Cancer.gov

Usual Intake of Total protein foods including beans and peas Table A20. Total protein foods including beans and peas: Means, percentiles and standard errors of usual intake, 2007-2010 Age (Years) N1 oz equivalents3 Mean (SE)2 5% (SE) 10% (SE) 25% (SE) 50%

51

Hematology, Serum Chemistry, and Serum Protein Electrophoresis Ranges for Free-ranging Roe Deer (Capreolus capreolus) in Sweden.  

PubMed

Abstract We present the first reference ranges for hematology (n?=?35 animals), serum biochemistry (n?=?62), and serum protein electrophoresis (n?=?32) in physically restrained free-ranging roe deer (Capreolus capreolus). Animals were captured in box traps and physically restrained for blood sampling during the winter in Sweden, 2011-13. No clinically significant sex or age differences were found. PMID:25375949

Küker, Susanne; Huber, Nikolaus; Evans, Alina; Kjellander, Petter; Bergvall, Ulrika A; Jones, Krista L; Arnemo, Jon M

2015-01-01

52

Drosophila hemolymph proteins: Purification, characterization, and genetic mapping of larval serum protein 2 in D. melanogaster  

Microsoft Academic Search

Three of the major protein species present in the hemolymph of Drosophila melanogaster larvae just prior to pupation are absent from second instar larvae but accumulate rapidly during the third instar. This article describes the purification and characterization of one of these, larval serum protein (LSP) 2, using an immunological assay. It is a homohexamer of molecular weight about 450,000,

Michael E. Akam; David B. Roberts; Jonathan Wolfe

1978-01-01

53

Binding of rat serum phosphorylcholine binding protein to platelets.  

PubMed

Rat serum phosphorylcholine binding protein (PCBP), a normal component of rat serum, inhibits in vitro aggregation of rat, rabbit and human platelets by interacting with platelets. In the present study, we have demonstrated the calcium-dependent, specific and saturable binding of 125I-PCBP to rat, rabbit and human platelets. Scatchard analysis of the binding data reveal a class of specific high-affinity binding sites with Kd values of 45.2 +/- 14.9, 26.1 +/- 8.3 and 32.2 +/- 9.9 nM on rat, rabbit and human platelets, respectively. These platelets also expressed a high capacity for binding to 125I-PCBP. The binding of 125I-PCBP to platelets was calcium- and time-dependent, and could be inhibited by phosphorylcholine (IC50 = 5.6 microM). Occupation of these binding sites by PCBP may be responsible for inhibition of platelet aggregation. PMID:2364084

Randell, E; Mookerjea, S; Nagpurkar, A

1990-06-20

54

High-abundance proteins depletion for serum proteomic analysis: concomitant removal of non-targeted proteins.  

PubMed

In clinical and pharmaceutical proteomics, serum and plasma are frequently used for detection of early diagnostic biomarkers for therapeutic targets. Although obtaining these body fluid samples is non-invasive and easy, they contain some abundant proteins that mask other protein components present at low concentrations. The challenge in identifying serum biomarkers is to remove the abundant proteins, uncovering and enriching at the same time the low-abundance ones. The depletion strategies, however, could lead to the concomitant removal of some non-targeted proteins that may be of potential interest. In this study, we compared three different methods aimed to deplete high-abundance proteins from human serum, focusing on the identification of non-specifically bound proteins which might be eventually removed. A Cibacron blue-dye-based method for albumin removal, an albumin and IgG immunodepletion method and an immunoaffinity column (Multiple Affinity Removal System) that simultaneously removes a total of six high-abundance proteins, were investigated. The bound proteins were eluted, separated by two-dimensional gel electrophoresis and identified by Nano LC-CHIP-MS system. Flow-through fractions and bound fractions were also analysed with the ProteinChip technology SELDI-TOF-MS. Our results showed that the methods tested removed not only the targeted proteins with high efficiency, but also some non-targeted proteins. We found that the Multiple Affinity Removal Column improved the intensity of low-abundance proteins, displayed new protein spots and increased resolution. Notably, the column showed the lowest removal of untargeted proteins, proved to be the most promising depletion approach and a reliable method for serum preparation prior to proteomic studies. PMID:20495836

Bellei, Elisa; Bergamini, Stefania; Monari, Emanuela; Fantoni, Luca Isaia; Cuoghi, Aurora; Ozben, Tomris; Tomasi, Aldo

2011-01-01

55

Influence of liposome charge and composition on their interaction with human blood serum proteins  

Microsoft Academic Search

Lipid composition and specially their electrostatic properties, were found to greatly influence the stability of liposomes in human blood serum. The amount and type of serum proteins bound to the liposomes were also clearly influenced by lipid composition and charge of liposomes. a good correlation was found between the amount of serum proteins adsorbed to a given type of liposome

Trinidad Hernfindez-Caselles; José Villalaín; Juan C. Gómez-Fernández

1993-01-01

56

Is serum S100B protein an useful biomarker in migraine?  

PubMed

Experimental data have demonstrated a role for S100B protein through the release of proinflammatory cytokines, following trigeminal nerve activation, implicated in the pathology of migraine. We investigated serum levels of S100B protein, as a peripheral glial biomarker, in patients with migraine. In total, 49 migraineurs and 35 age- and gender-matched controls were enrolled in this prospective clinical study. The migraine diagnosis was made according to the International Classification of Headache Disorders II diagnostic criteria. Serum samples were obtained for the measurement of S100B levels from all participants and were analyzed using commercial enzyme-linked immunosorbent assay kits. Serum S100B levels were significantly lower in migraineurs than controls (p < 0.001). S100B levels did not significantly differ in migraineurs with or without aura (p > 0.05). In addition, there was no correlation between serum S100B levels and headache characteristics, including attack severity, frequency and duration, and disease duration (p > 0.05). These findings suggest that serum S100B levels were significantly decreased in migraine patients, but further research is needed to ascertain the contribution of S100B in the clinical evaluation of migraine. PMID:24531979

Celikbilek, Asuman; Sabah, Seda; Tanik, Nermin; Ak, Hakan; Atalay, Tugay; Yilmaz, Neziha

2014-08-01

57

Evaluation of serum protein electrophoresis in greater rhea ( Rhea americana Linnaeus, 1758)  

Microsoft Academic Search

Serum proteins from 47 healthy greater rheas (Rhea americana; male and female) were separated by electrophoresis in order to characterize normal reference ranges. Determination of total\\u000a protein concentration was performed through biuret reaction. The mean value of total serum protein was 4.4 g\\/dL. Absolute\\u000a concentrations of serum proteins were determined by agarose gel electrophoretic fractioning. Five fractions were analyzed:\\u000a albumin, alpha,

Cybele Esteves Almeida; Barbara Charlotte Bach; Maristela Lovato Flores; Rogério Pereira Fontoura; Stefanie Dickel Segabinazi; Marta Helena Carlesso Aita

2010-01-01

58

Embryo culture in teratological surveillance and serum proteins in development. Progress report, 1979-1980  

SciTech Connect

Research progress for the period 1979-1980 is reported. The feasibility of using rat embryo cultures to test the teratogenic activity of serum was studied. The mechanisms regulating the synthesis of serum proteins were investigated. (ACR)

Klein, N.W.

1980-07-01

59

Bayesian hierarchical reconstruction of protein profiles including a digestion model  

E-print Network

Bayesian hierarchical reconstruction of protein profiles including a digestion model Pierre to recover the protein biomarkers content in a robust way. We will focus on the digestion step since and each branch to a molecular processing such as digestion, ionisation and LC-MS separation

Paris-Sud XI, Université de

60

Evaluation of Plasma Fibrinogen Degradation Products and Total Serum Protein Concentration in Oral Submucous Fibrosis  

PubMed Central

Background: Oral submucous fibrosis (OSMF) is a potentially malignant disorder with a multifactorial etiology. Malnutrition is a major problem for the inhabitants of most countries where OSMF is prevalent. Recently, a new direction in the etiopathogenesis was provided by the identification of fibrinogen degradation products (FDP) in the plasma of OSMF patients. Aims and Objectives: To assess the role of FDP in the etiology of OSMF and to correlate with the nutritional status by evaluating the total serum protein level. The study also determines to evaluate the correlation between the levels of plasma FDP with respect to the staging and grading of OSMF. Correlation between the levels of Total Serum Protein (TSP) with respect to the staging and grading of OSMF was also evaluated. Materials and Methods: The study included 30 cases clinically and histopathologically diagnosed as oral submucous fibrosis. The FDP levels were assessed using both qualitative and semi quantitative method as supplied by ‘Tulip Diagnostics (P) Ltd. Total Serum Protein (TSP) estimation was done by Biuret method using Liquixx Protein kit by Erba, Manheim. Results: The study indicates that in qualitative assessment of FDP only 14 subjects showed the presence of FDP levels>200ng/ml. In semiquantitative assessment there is no significant association between varying clinical stages and histopathological grades and FDP levels. Total serum Protein level showed a marginal increase in all subjects. The study revealed a positive correlation between FDP and TSP in all OSMF subjects. Conclusion: A larger sample size which would be a better representation of the population and the use of different methods which have higher sensitivities and specificities to evaluate FDP level and detailed fractional analysis of protein along with immunoglobulin profiling would facilitate in attaining more conclusive results. PMID:24995245

B.N.V.S., Satish; B., Maharudrappa; K.M., Prashant; Hugar, Deepa; Allad, Umesh; Prabhu, Prasanth S.

2014-01-01

61

A Sex-Limited Serum Protein of Syrian Hamsters: Definition of Female Protein and Regulation by Testosterone  

Microsoft Academic Search

Normal Syrian hamster females contain a serum protein not found by simple gel diffusion assay in normal adult males. This sex-limited protein was called female protein (FP). Low levels of FP were found in sera from normal weanling male hamsters. Adult male hamsters castrated or treated with diethylstilbestrol also developed serum FP, which could be suppressed by administration of testosterone.

J. E. Coe

1977-01-01

62

Changes in concentration and fractions of blood serum proteins of chickens during fattening  

Microsoft Academic Search

This paper describes research into concentration of total proteins and individual protein fractions in the blood serum of Ross broiler chickens during fattening period. Blood for analysis was taken at the 14th, 28th and 42 nd days of age by wing vein puncture. Concentration of total serum proteins was detected by spectrophotometry. Electrophoresis on gelled cellulose-acetate tapes determined the shares

Suzana Milinkovi?-Tur; Maja Zdelar-Tuk

2007-01-01

63

Reference intervals for acute phase protein and serum protein electrophoresis values in captive Asian elephants (Elephas maximus).  

PubMed

Acute phase protein (APP) immunoassays and serum protein electrophoresis (SPEP) are assays for evaluating the inflammatory response and have use as diagnostic tools in a variety of species. Acute phase proteins are markers of inflammation that are highly conserved across different species while SPEP separates and quantifies serum protein fractions based on their physical properties. In the current study, serum samples from 35 clinically healthy Asian elephants (Elephas maximus) were analyzed using automated assays for C-reactive protein, serum amyloid A, and haptoglobin and SPEP. Robust methods were used to generate reference intervals for the APPs: C-reactive protein (1.3-12.8 mg/l), serum amyloid A (0-47.5 mg/l), and haptoglobin (0-1.10 mg/ml). In addition, SPEP was performed on these samples to establish reference intervals for each protein fraction. A combination of APPs and SPEP measurements are valuable adjunctive diagnostic tools in elephant health care. PMID:25057161

Isaza, Ramiro; Wiedner, Ellen; Hiser, Sarah; Cray, Carolyn

2014-09-01

64

Clinical Utility of Serum Autoantibodies Detected by Protein Microarray in Melanoma  

PubMed Central

Better prognostic and predictive markers in melanoma are needed to select patients for therapy. We utilized a dual-lectin affinity chromatography and a natural protein microarray-based analysis to select a subproteome of target glycoproteins to profile serum antibodies against melanoma associated antigens that may predict nodal positivity. We identified 5 melanoma-associated antigens using this microarray coupled to mass spectrometry; GRP75, GRP94, ASAH1, CTSD and LDHB. We evaluated their predictive value for nodal status adjusting for age, gender, Breslow thickness, mitotic rate and ulceration using standard logistic regression. After adjustment, ASAH1, CTSD and LDHB were significantly negatively associated with nodal status (P = 0.0008) and GRP94 was significantly positively associated (P = 0.014). Our best multivariate model for nodal positivity included Breslow thickness, presence of serum anti-ASAH1, anti-LDHB or anti-CTSD, and presence of serum anti-GRP94, with an area under the ROC curve of 0.869. If validated, these results show promise for selecting clinically node negative patients for SLN biopsy. In addition, there is strong potential for glycoprotein microarray to screen serum autoantibodies that may identify patients at high risk of distant metastases or those likely or unlikely to respond to treatment, and these proteins may serve as targets for intervention. PMID:22084687

Sabel, Michael S.; Liu, Yashu; Griffith, Kent A.; He, Jintang; Xie, Xaiolei; Lubman, David M.

2011-01-01

65

Targeted quantification of low ng/mL level proteins in human serum without immunoaffinity depletion  

SciTech Connect

We recently reported an antibody-free targeted protein quantification strategy, termed high-pressure, high-resolution separations with intelligent selection and multiplexing (PRISM) for achieving significantly enhanced sensitivity using selected reaction monitoring (SRM) mass spectrometry. Integrating PRISM with front-end IgY14 immunoaffinity depletion, sensitive detection of targeted proteins at 50-100 pg/mL levels in human blood plasma/serum was demonstrated. However, immunoaffinity depletion is often associated with undesired losses of target proteins of interest. Herein we report further evaluation of PRISM-SRM quantification of low-abundance serum proteins without immunoaffinity depletion and the multiplexing potential of this technique. Limits of quantification (LOQs) at low ng/mL levels with a median CV of ~12% were achieved for proteins spiked into human female serum using as little as 2 µL serum. PRISM-SRM provided up to ~1000-fold improvement in the LOQ when compared to conventional SRM measurements. Multiplexing capability of PRISM-SRM was also evaluated by two sets of serum samples with 6 and 21 target peptides spiked at the low attomole/µL levels. The results from SRM measurements for pooled or post-concatenated samples were comparable to those obtained from individual peptide fractions in terms of signal-to-noise ratios and SRM peak area ratios of light to heavy peptides. PRISM-SRM was applied to measure several ng/mL-level endogenous plasma proteins, including prostate-specific antigen, in clinical patient sera where correlation coefficients > 0.99 were observed between the results from PRISM-SRM and ELISA assays. Our results demonstrate that PRISM-SRM can be successfully used for quantification of low-abundance endogenous proteins in highly complex samples. Moderate throughput (50 samples/week) can be achieved by applying the post-concatenation or fraction multiplexing strategies. We anticipate broad applications for targeted PRISM-SRM quantification of low-abundance cellular proteins in systems biology studies as well as candidate biomarkers in biofluids.

Shi, Tujin; Sun, Xuefei; Gao, Yuqian; Fillmore, Thomas L.; Schepmoes, Athena A.; Zhao, Rui; He, Jintang; Moore, Ronald J.; Kagan, Jacob; Rodland, Karin D.; Liu, Tao; Liu, Alvin Y.; Smith, Richard D.; Tang, Keqi; Camp, David G.; Qian, Weijun

2013-07-05

66

Acoustical method and device for determination of lipid and protein spectra of blood serum  

E-print Network

Acoustical method and device for determination of lipid and protein spectra of blood serum I fractions and lipid components of the human blood serum are presented. Acoustic methods are based on high absorption. Acoustic characteristics of the blood serum were measured using the method of a fixed length

Paris-Sud XI, Université de

67

Dye-promoted precipitation of serum proteins. Mechanism and application.  

PubMed

Immobilized dyes have been used primarily for purification of nucleotide dependent enzymes and proteins from plasma and other sources. Due to their low costs, high protein binding capacity and resistance to degradation dyes bear the potential as ligand for affinity separation of proteins on a large scale. In this paper dyes have been used for precipitation of proteins. Using albumin, prealbumin, alpha 1-acid glycoprotein and immunoglobulin G as model proteins we could demonstrate that dye-promoted precipitation depends on several factors which include the structure of the dye, the pH of the solution, the dye/protein molar ratio and the intrinsic properties of the proteins. It revealed that most of the dyes tested were endowed with the precipitating potential. The efficacy of precipitation was found to increase with the complexity of the dye structure. However, the amount of a dye required for total precipitation was found to be different for a given protein. Electrostatic as well as hydrophobic forces are involved in the mechanism of precipitation. It was demonstrated that by optimizing the conditions, mixtures of proteins can be resolved by dye-promoted precipitation. The high sensitivity of the reaction offers the possibility of using this method for rapid concentration of very diluted protein solutions. PMID:1367693

Birkenmeier, G; Kopperschläger, G

1991-11-01

68

Serum Monocyte Chemoattractant Protein-1 in Pancreatic Cancer  

PubMed Central

Background/Aims. Pancreatic ductal adenocarcinoma (PDA) has etiological association with chronic inflammation. Elevated circulating levels of inflammatory mediators, such as monocyte chemoattractant protein-1 (MCP-1), are found in obese individuals. We hypothesized that serum MCP-1 levels are elevated in obese PDA patients. Methods. ELISA was used to analyze MCP-1 serum levels in PDA (n = 62) and intraductal papillary mucinous neoplasms (IPMN) (n = 27). Recursive partitioning statistical analysis investigated the relationship between log MCP-1 and clinicopathological parameters. Results. Log MCP-1 values were significantly (P < 0.05) elevated in patients with BMI ? 37.5. In patients with BMI < 37.5, average log MCP-1 values were significantly elevated in PDA patients when compared to IPMN patients. Within the IPMN group, higher log MCP-1 levels correlated with increased age. Recursive partitioning analysis of IPMN versus PDA revealed a strategy of predicting characteristics of patients who are more likely to have cancer. This strategy utilizes log MCP-1 as the primary factor and also utilizes smoking status, gender, and age. Conclusion. MCP-1 is a promising biomarker in pancreatic cancer. The potential of using MCP-1 to distinguish PDA from IPMN patients must be studied in larger populations to validate and demonstrate its eventual clinical utility. PMID:21977031

Sullivan, Jennifer; Gong, Qiaoke; Hyslop, Terry; Lavu, Harish; Chipitsyna, Galina; Yeo, Charles J.; Arafat, Hwyda A.

2011-01-01

69

A PHASE RULE STUDY OF THE PROTEINS OF THE BLOOD SERUM: A COMPARISON OF THE PROTEINS OF HUMAN, RAT, AND HORSE SERUM  

PubMed Central

There are four different kinds of protein in blood serum as shown by the solubility curves. They must be either single proteins, several continuous series of compounds, or solid solutions. The solid protein phases are hydrated. There are definite sex and species differences. PMID:19873004

Jameson, Eloise; Roberts, Dorothy Brown

1937-01-01

70

Using experimental data designs and multivariate modeling to assess the effect of glycated serum protein concentration on glucose prediction from near-infrared spectra of human serum.  

PubMed

Near-infrared (NIR) spectra of human blood serum consist of overlapping strong absorption bands of water and serum proteins, which affect the ability of multivariate calibration models to predict glucose. Furthermore, serum proteins such as albumin and globulins undergo a glycation reaction by forming covalent bonds with freely available glucose molecules in the serum. In diabetic individuals with poor glucose control, more and more serum protein molecules react with glucose, resulting in a high glycated protein concentration. The glucose molecules covalently bonded to serum proteins might contribute to the overall glucose signal acquired by NIR spectroscopy. This might affect the prediction ability of multivariate calibration models such as partial least squares regression (PLSR). In this study, we investigated the effect of total protein concentration and the glycated protein concentration in blood serum on the prediction ability of PLSR calibration models. Serum samples were subjected to ultra-filtration, and the PLSR model was built using NIR spectra of filtered serum solutions. Prediction performance was found to improve by 39-42% in absence of serum protein molecules. Various experimental data set designs were generated by carefully varying the glycated serum protein concentration in calibration and test sets of PLSR models. This investigation revealed that the impact of varying glycated protein concentration on the root mean square error of prediction was not drastic. To test the statistical significance of the prediction results, a multiple linear regression model was built. The glycated serum protein concentration was found to be statistically insignificant (p = 0.86) in predicting glucose concentration. Overall, it was concluded that the glycated serum proteins do not affect the glucose prediction accuracy of PLSR models using NIR spectra of human serum. PMID:24694695

Sharma, Sandeep; Goodarzi, Mohammad; Delanghe, Joris; Ramon, Herman; Saeys, Wouter

2014-01-01

71

Birmingham Medical Research Expeditionary Society 1977 Expedition: serum and urine proteins during a high altitude trek  

Microsoft Academic Search

Serum and urine proteins were measured daily in 17 subjects undertaking a typical high altitude Himalayan trek. Marked changes occurred in a variety of serum proteins as a result of plasma volume alterations and 'stress'. There was only a sporadic increase in proteinuria. None of the changes was related to the development of acute mountain sickness.

A. R. Bradwell

1979-01-01

72

Autocrine Induction of Collagenase by Serum Amyloid ALike and beta 2-microglobulin--Like Proteins  

Microsoft Academic Search

Two autocrine proteins of 14 and 12 kilodaltons that induce the synthesis of rabbit fibroblast collagenase were identified. The proteins were purified from serum-free culture medium taken from rabbit synovial fibroblasts stimulated with phorbol myristate acetate. The amino-terminal sequences of the 14- and 12-kilodalton species were ~ 60 to 80 percent homologous with serum amyloid A and beta 2 microglobulin,

Constance E. Brinckerhoff; Teresa I. Mitchell; Michael J. Karmilowicz; Barbara Kluve-Beckerman; Merrill D. Benson

1989-01-01

73

Influence of protein intake associated with coconut or salmon oil on serum,  

E-print Network

Influence of protein intake associated with coconut or salmon oil on serum, VLDL, LDL and HDL of protein depletion associated with salmon (rich in w3 PUFA) or co- conut oil (poor in EFA) on various serum% casein + 5% salmon oil), SAd (2% casein + 5% salmon oil), COC (20% casein + 5% coconut oil), COd (2

Paris-Sud XI, Université de

74

Automated method for the estimation of serum protein-bound iodine following alkaline incineration  

PubMed Central

A method is described for the estimation of serum protein-bound iodine using alkaline incineration and an automated technique for the estimation of iodine in the ash. Pretreatment of the serum with an anion exchange resin avoids the need for precipitation and washing of the protein. The method is accurate, reproducible, and simple to perform. PMID:5919368

Welshman, S. G.; Bell, J. F.; McKee, G.

1966-01-01

75

Sex-Associated Differences in Serum Proteins of Mice  

Microsoft Academic Search

Agar electrophoresis of serum from mice of C57BL\\/10-H-2d, B10.Sn, A.SW, A.CA, R III, and P1 inbred strains shows that the females have a lower concentration of alpha -1 serum globulin than the males and, in some strains, the females also have a lower concentration of alpha -2 and beta -serum globulin. Females of the A.SW strain have a higher serum

E. Espinosa; E. Canelo; M. Bravo; O. Gonzalez

1964-01-01

76

Relationship between serum growth hormone binding protein levels and height in young men.  

PubMed

The biochemical mediators responsible for variations in stature among normal subjects are largely unknown. To obtain some initial information about potential endocrine factors, we measured the serum concentrations of GH, IGF-1, IGFBP-3 and GHBP in healthy young men shorter than 159 cm and taller than 187 cm. We studied 14 volleyball and basketball players (tall group), and 14 jockey students from a horse racetrack (short group). A careful medical history was taken, including dietary intake, and physical examination with special attention to the possible presence of genetic stigmata was performed. Serum prealbumin was determined as an index of nutritional status. A buccal smear was performed to exclude Klinefelter's syndrome. The BMI and serum prealbumin levels were comparable in both groups of individuals. The nutritional survey, however, revealed that the tall subjects had a higher intake of calories (42.2+/-11.2 vs. 30.1+/-15.15 kcal/kg, p<0.05), and protein (1.5+/-0.6 vs. 0.8+/-0.4 mg/kg, p<0.01). Serum concentrations of GHBP did not differ in the two groups (0.95+/-0.37 nmol/l in the tall, and 0.95+/-0.53 nmol/l in the short group), and did not correlate with height, serum IGF-I levels, or BMI. We observed a significant difference in the serum concentrations of IGF-I in the two groups of individuals (42.02+/-9.37 nmol/l in the tall and 31.79+/-3.18 nmol/l in the short group, p<0.05), and this growth factor showed a positive correlation with height (r = 0.5, p<0.01). These preliminary findings suggest that final height differences in young men do not appear to be mediated by variations in GHBP concentrations. PMID:10968476

Codner, E; Mericq, M V; Maheshwari, H G; Iñguez, G; Capurro, M T; Salazar, T; Baumann, G; Cassorla, F; Codner, D E

2000-01-01

77

Serum antibody response to recombinant major inner capsid protein following human infection with group B rotavirus.  

PubMed Central

Recombinant major inner capsid protein (VP6) of the IDIR strain of group B rotavirus (GBR) was incorporated in a solid-phase immunoassay to access antibody response to infection in humans. Expression of VP6 in insect cells permitted design of a highly sensitive assay that avoided the contaminants present in GBR antigens obtained from fecal specimens. Among patients infected with the ADRV strain of GBR in China, increased reactivity with recombinant VP6 was observed in convalescent-phase sera in comparison with sera obtained shortly after infection (P = 0.0084). Anti-VP6 antibodies were detectable as soon as 7 days after onset of gastrointestinal symptoms, and serum reactivity persisted in specimens drawn more than 1 year after infection. Solid-phase immunoassay with recombinant VP6 was next employed in order to assess anti-GBR antibody in 513 serum specimens obtained from 423 Maryland residents (ages, 7 months to 96 years; median age, 42 years). Four individuals (< 1%) exhibited serum antibodies directed against the recombinant VP6 (ages, 54 to 95 years; mean age, 77 years). Examination of 129 additional serum specimens including some from other geographic regions of the United States failed to reveal the presence of anti-GBR antibody. Anti-GBR antibody was also not detected in any of 131 serum specimens from 60 staff and residents of a nursing home in Switzerland. While infection of humans with GBR has been uncommon in these locations outside of China, the detection of serum antibodies in older individuals in the United States either indicated an unknown, age-related risk factor or may have indicated infection in the more distant past. The availability of these reagents should allow surveys for GBR infection among additional populations that have not previously been investigated. PMID:8077413

Eiden, J J; Mouzinho, A; Lindsay, D A; Glass, R I; Fang, Z Y; Taylor, J L

1994-01-01

78

Serum antibody response to recombinant major inner capsid protein following human infection with group B rotavirus.  

PubMed

Recombinant major inner capsid protein (VP6) of the IDIR strain of group B rotavirus (GBR) was incorporated in a solid-phase immunoassay to access antibody response to infection in humans. Expression of VP6 in insect cells permitted design of a highly sensitive assay that avoided the contaminants present in GBR antigens obtained from fecal specimens. Among patients infected with the ADRV strain of GBR in China, increased reactivity with recombinant VP6 was observed in convalescent-phase sera in comparison with sera obtained shortly after infection (P = 0.0084). Anti-VP6 antibodies were detectable as soon as 7 days after onset of gastrointestinal symptoms, and serum reactivity persisted in specimens drawn more than 1 year after infection. Solid-phase immunoassay with recombinant VP6 was next employed in order to assess anti-GBR antibody in 513 serum specimens obtained from 423 Maryland residents (ages, 7 months to 96 years; median age, 42 years). Four individuals (< 1%) exhibited serum antibodies directed against the recombinant VP6 (ages, 54 to 95 years; mean age, 77 years). Examination of 129 additional serum specimens including some from other geographic regions of the United States failed to reveal the presence of anti-GBR antibody. Anti-GBR antibody was also not detected in any of 131 serum specimens from 60 staff and residents of a nursing home in Switzerland. While infection of humans with GBR has been uncommon in these locations outside of China, the detection of serum antibodies in older individuals in the United States either indicated an unknown, age-related risk factor or may have indicated infection in the more distant past. The availability of these reagents should allow surveys for GBR infection among additional populations that have not previously been investigated. PMID:8077413

Eiden, J J; Mouzinho, A; Lindsay, D A; Glass, R I; Fang, Z Y; Taylor, J L

1994-06-01

79

Identification of anti-adipogenic proteins in adult bovine serum suppressing 3T3-L1 preadipocyte differentiation  

PubMed Central

Adipocyte differentiation is a complex developmental process forming adipocytes from various precursor cells. The murine 3T3-L1 preadipocyte cell line has been most frequently used in the studies of adipocyte differentiation. Differentiation of 3T3-L1 preadipocytes includes a medium containing fetal bovine serum (FBS) with hormonal induction. In this study, we observed that differentiation medium containing adult bovine serum (ABS) instead of FBS did not support differentiation of preadipocytes. Impaired adipocyte differentiation was due to the presence of a serum protein factor in ABS that suppresses differentiation of preadipocytes. Using a proteomic analysis, alpha-2-macroglobulin and paraoxonase/arylesterase 1, which were previously shown to suppress differentiation of preadipocytes, were identified as anti-adipogenic proteins. Although their functional mechanisms have not yet been elucidated, the anti-adipogenic effects of these proteins are discussed. [BMB Reports 2013; 46(12): 582-587] PMID:24195790

Park, Jeongho; Park, Jihyun; Nahm, Sang-Soep; Choi, Inho; Kim, Jihoe

2013-01-01

80

Adsorption and adhesion of common serum proteins to nanotextured gallium nitride.  

PubMed

As the broader effort towards device and material miniaturization progresses in all fields, it becomes increasingly important to understand the implications of working with functional structures that approach the size scale of molecules, particularly when considering biological systems. It is well known that thin films and nanostructures feature different optical, electrical, and mechanical properties from their bulk composites; however, interactions taking place at the interface between nanomaterials and their surroundings are less understood. Here, we explore interactions between common serum proteins - serum albumin, fibrinogen, and immunoglobulin G - and a nanotextured gallium nitride surface. Atomic force microscopy with a carboxyl-terminated colloid tip is used to probe the 'activity' of proteins adsorbed onto the surface, including both the accessibility of the terminal amine to the tip as well as the potential for protein extension. By evaluating the frequency of tip-protein interactions, we can establish differences in protein behaviour on the basis of both the surface roughness as well as morphology, providing an assessment of the role of surface texture in dictating protein-surface interactions. Unidirectional surface features - either the half-unit cell steppes of as-grown GaN or those produced by mechanical polishing - appear to promote protein accessibility, with a higher frequency of protein extension events taking place on these surfaces when compared with less ordered surface features. Development of a full understanding of the factors influencing surface-biomolecule interactions can pave the way for specific surface modification to tailor the bio-material interface, offering a new path for device optimization. PMID:25564044

Bain, Lauren E; Hoffmann, Marc P; Bryan, Isaac; Collazo, Ramón; Ivanisevic, Albena

2015-01-28

81

Two major ruminant acute phase proteins, haptoglobin and serum amyloid A, as serum biomarkers during active sheep scab infestation  

PubMed Central

Two ruminant acute phase proteins (APPs), haptoglobin (Hp) and serum amyloid A (SAA), were evaluated as serum biomarkers (BMs) for sheep scab–a highly contagious ectoparasitic disease caused by the mite Psoroptes ovis, which is a major welfare and production threat worldwide. The levels of both APPs increased in serum following experimental infestation of sheep with P. ovis, becoming statistically significantly elevated from pre-infestation levels at 4 weeks post-infestation. Following successful treatment of infested sheep with an endectocide, Hp and SAA serum levels declined rapidly, with half lives of less than 3 days. In contrast, serum IgG levels which specifically bound the P. ovis-derived diagnostic antigen Pso o 2 had a half-life of 56 days. Taking into account pre-infestation serum levels, rapidity of response to infestation and test sensitivity at the estimated optimum cut-off values, SAA was the more discriminatory marker. These studies illustrated the potential of SAA and Hp to indicate current sheep scab infestation status and to augment the existing Pso o 2 serological assay to give disease-specific indications of both infestation and successful treatment. PMID:24176040

2013-01-01

82

Clinical impact of serum proteins on drug delivery.  

PubMed

Among serum proteins albumin and transferrin have attracted the most interest as drug carriers in the past two decades. Prior to that, their potential use was overshadowed by the advent of monoclonal antibodies that was initiated by Milstein and Koehler in 1975. Meanwhile intensive pursuit of exploiting transferrin, but above all albumin as an exogenous or endogenous carrier protein for treating various diseases, primarily cancer, rheumatoid arthritis, diabetes and hepatitis has resulted in several marketed products and numerous clinical trials. While the use of transferrin has clinically been primarily restricted to immunotoxins, albumin-based drug delivery systems ranging from albumin drug nanoparticles, albumin fusion protein, prodrugs and peptide derivatives that bind covalently to albumin as well as physically binding antibody fragments and therapeutically active peptides are in advanced clinical trials or approved products. For treating diabetes, Levemir and Victoza that are myristic acid derivatives of human insulin or glucagon-like peptide 1 (GLP-1) act as long-acting peptides by binding to the fatty acid binding sites on circulating albumin to control glucose levels. Levemir from Novo Nordisk has already developed into a blockbuster since its market approval in 2004. Abraxane, an albumin paclitaxel nanoparticle as a water-soluble galenic formulation avoiding the use of cremophor/ethanol, transports paclitaxel through passive targeting as an albumin paclitaxel complex to the tumor site and is superior to conventional Taxol against metastatic breast cancer. INNO-206, an albumin-binding doxorubicin prodrug that also accumulates in solid tumors due to the enhanced permeability and retention (EPR) effect but releases the parent drug through acid cleavage, either intra- or extracellularly, is entering phase II studies against sarcoma. An expanding field is the use of albumin-binding antibody moieties which do not contain the fragment crystallizable (Fc) portion of, conventional immunoglobulin G (IgG) but are comprised of monovalent or bivalent light and/or heavy chains and incorporate an additional albumin-binding peptide or antibody domain. The most advanced antibody of this kind is ATN-103 (Ozoralizumab), a trivalent albumin-binding nanobody that neutralizes the pro-inflammatory tumor necrosis factor alpha (TNF-?) as a causative agent for exacerbating rheumatoid arthritis. ATN-103 is currently in multi-center phase II trials against this debilitating disease. In summary, because albumin as the most abundant circulating protein cannot only be used to improve the pharmacokinetic profile of therapeutically relevant peptides and the targeting moiety of antibodies but also for peptide-based targeting as well as low-molecular weight drugs to inflamed or malignant tissue, it is anticipated that R&D efforts of academia and the pharmaceutical industry in this field of drug delivery will prosper. PMID:22155554

Kratz, Felix; Elsadek, Bakheet

2012-07-20

83

Smoking and serum proteins in atomic-bomb survivors in Japan  

SciTech Connect

Associations of smoking habit with serum levels of total protein as well as protein fractions were studied in a population consisting of 4,739 atomic-bomb survivors and unexposed control subjects in Hiroshima, Japan who participated in the 1979-1981 period of the Adult Health Study, an ongoing health follow-up program of the Radiation Effects Research Foundation. Smoking was strongly related to serum protein concentration after correction for age, sex, and body mass index. Among current smokers, levels of total protein, beta globulin, and gamma globulin were significantly lower and levels of alpha-1 and alpha-2 globulin were significantly higher, when compared with nonsmokers. For serum albumin levels a decrease was also noted, but it failed to attain statistical significance. Ex-smokers were indistinguishable from nonsmokers in terms of the serum protein levels analyzed. With an increase of the amount of daily cigarette consumption, monotonic increases of serum levels were observed only in alpha-1 globulin. Duration of smoking was related to increased alpha-1 and alpha-2 globulin. Smoking duration was also associated with albumin level, but the trend was not monotonic. The radiation exposure effect on serum protein level was significant in several instances but was in general much smaller than the smoking effect, and its inclusion in the regression models did not noticeably affect the association between smoking and serum proteins.

Stram, D.O.; Akiba, S.; Neriishi, K.; Stevens, R.G.; Hosoda, Y. (Univ. of Southern California, Pasadena (USA))

1990-06-01

84

Serum protein capillary electrophoresis and measurement of acute phase proteins in a captive cheetah (Acinonyx jubatus) population.  

PubMed

Renal and gastrointestinal pathologies are widespread in the captive cheetah (Acinonyx jubatus) population but are often diagnosed at a late stage, because diagnostic tools are limited to the evaluation of clinical signs or general blood examination. Presently, no data are available on serum proteins and acute-phase proteins in cheetahs during health or disease, although they might be important to improve health monitoring. This study aimed to quantify serum proteins by capillary electrophoresis in 80 serum samples from captive cheetahs, categorized according to health status and disease type. Moreover, serum amyloid A concentrations were measured via a turbidimetric immunoassay validated in domestic cats, whereas haptoglobin and C-reactive protein were determined by non-species-specific functional tests. Cheetahs classified as healthy had serum protein and acute phase protein concentrations within reference ranges for healthy domestic cats. In contrast, unhealthy cheetahs had higher (P < 0.001) serum amyloid A, alpha2-globulin, and haptoglobin concentrations compared with the healthy subgroup. Moreover, serum amyloid A (P = 0.020), alpha2-globulin (P < 0.001) and haptoglobin (P = 0.001) concentrations in cheetahs suffering from chronic kidney disease were significantly greater compared to the reportedly healthy cheetahs. Our study indicates that serum proteins in the cheetah can be analyzed by routine capillary electrophoresis, whereas acute-phase proteins can be measured using available immunoassays or non-species-specific techniques, which are also likely to be applicable in other exotic felids. Moreover, results suggest that serum amyloid A and haptoglobin are important acute-phase proteins in the diseased cheetah and highlight the need to evaluate their role as early-onset markers for disease. PMID:25314816

Depauw, Sarah; Delanghe, Joris; Whitehouse-Tedd, Katherine; Kjelgaard-Hansen, Mads; Christensen, Michelle; Hesta, Myriam; Tugirimana, Pierrot; Budd, Jane; Dermauw, Veronique; Janssens, Geert P J

2014-09-01

85

Disparate proteome responses of pathogenic and nonpathogenic aspergilli to human serum measured by activity-based protein profiling (ABPP).  

PubMed

Aspergillus fumigatus is the primary pathogen causing the devastating pulmonary disease Invasive Aspergillosis in immunocompromised individuals. There is high genomic synteny between A. fumigatus and closely related rarely pathogenic Neosartorya fischeri and Aspergillus clavatus genomes. We applied activity-based protein profiling to compare unique or overexpressed activity-based probe-reactive proteins of all three fungi over time in minimal media growth and in response to human serum. We found 360 probe-reactive proteins exclusive to A. fumigatus, including known virulence associated proteins, and 13 proteins associated with stress response exclusive to A. fumigatus culture in serum. Though the fungi are highly orthologous, A. fumigatus has a significantly greater number of ABP-reactive proteins across varied biological process. Only 50% of expected orthologs of measured A. fumigatus reactive proteins were observed in N. fischeri and A. clavatus. Activity-based protein profiling identified a number of processes that were induced by human serum in A. fumigatus relative to N. fischeri and A. clavatus. These included actin organization and assembly, transport, and fatty acid, cell membrane, and cell wall synthesis. Additionally, signaling proteins regulating vegetative growth, conidiation, and cell wall integrity, required for appropriate cellular response to external stimuli, had higher activity-based probe-protein reaction over time in A. fumigatus and N. fisheri, but not in A. clavatus. Together, we show that measured proteins and physiological processes identified solely or significantly over-represented in A. fumigatus reveal a unique adaptive response to human protein not found in closely related, but rarely pathogenic aspergilli. These unique activity-based probe-protein responses to culture condition may reveal how A. fumigatus initiates pulmonary invasion leading to Invasive Aspergillosis. PMID:23599423

Wiedner, Susan D; Ansong, Charles; Webb-Robertson, Bobbie-Jo; Pederson, LeeAnna M; Fortuin, Suereta; Hofstad, Beth A; Shukla, Anil K; Panisko, Ellen A; Smith, Richard D; Wright, Aaron T

2013-07-01

86

Disparate Proteome Responses of Pathogenic and Nonpathogenic Aspergilli to Human Serum Measured by Activity-Based Protein Profiling (ABPP)*  

PubMed Central

Aspergillus fumigatus is the primary pathogen causing the devastating pulmonary disease Invasive Aspergillosis in immunocompromised individuals. There is high genomic synteny between A. fumigatus and closely related rarely pathogenic Neosartorya fischeri and Aspergillus clavatus genomes. We applied activity-based protein profiling to compare unique or overexpressed activity-based probe-reactive proteins of all three fungi over time in minimal media growth and in response to human serum. We found 360 probe-reactive proteins exclusive to A. fumigatus, including known virulence associated proteins, and 13 proteins associated with stress response exclusive to A. fumigatus culture in serum. Though the fungi are highly orthologous, A. fumigatus has a significantly greater number of ABP-reactive proteins across varied biological process. Only 50% of expected orthologs of measured A. fumigatus reactive proteins were observed in N. fischeri and A. clavatus. Activity-based protein profiling identified a number of processes that were induced by human serum in A. fumigatus relative to N. fischeri and A. clavatus. These included actin organization and assembly, transport, and fatty acid, cell membrane, and cell wall synthesis. Additionally, signaling proteins regulating vegetative growth, conidiation, and cell wall integrity, required for appropriate cellular response to external stimuli, had higher activity-based probe-protein reaction over time in A. fumigatus and N. fisheri, but not in A. clavatus. Together, we show that measured proteins and physiological processes identified solely or significantly over-represented in A. fumigatus reveal a unique adaptive response to human protein not found in closely related, but rarely pathogenic aspergilli. These unique activity-based probe-protein responses to culture condition may reveal how A. fumigatus initiates pulmonary invasion leading to Invasive Aspergillosis. PMID:23599423

Wiedner, Susan D.; Ansong, Charles; Webb-Robertson, Bobbie-Jo; Pederson, LeeAnna M.; Fortuin, Suereta; Hofstad, Beth A.; Shukla, Anil K.; Panisko, Ellen A.; Smith, Richard D.; Wright, Aaron T.

2013-01-01

87

Altered Phospholipid Transfer Protein Gene Expression and Serum Lipid Profile by Topotecan  

PubMed Central

Camptothecin (CPT) and its structural analogues including topotecan and irinotecan, are inhibitors of topoisomerase I. These drugs are clinically active against a broad spectrum of cancers. To understand the genesis of chemotherapeutic resistance to the CPT family of anticancer drugs, we examined by gene expression profiling the pharmacological response to topotecan in the human hepatoma HepG2 cells and found a striking induction of the phospholipid transfer protein (PLTP) gene expression by topotecan. We showed that activation of PLTP gene expression is specific to CPT and its analogues including specific enantiomers that inhibit topoisomerase I. PLTP-mediated lipid transfer to high-density lipoprotein (HDL) is thought to be important for shuttling and redistribution of lipids between lipoproteins, which are normally returned to the liver for metabolism via the reverse cholesterol transport pathway. Hence, we asked whether elevated PLTP levels might increase the transfer of drugs into HDL. We observed that CPT was not accumulated in HDL and other lipoproteins. In addition, topotecan treatment in mice caused a marked reduction in serum HDL that was accompanied by an increase in triglyceride and cholesterol levels. These results showed that PLTP does not mediate the transfer of topoisomerase I inhibitors to serum lipoproteins. However, elevated serum PLTP levels following treatment with topoisomerase I inhibitors in cancer patients may serve as a biomarker for monitoring the development of hypertriglyceridemia and acute pancreatitis. PMID:20416282

Saunders, Rudel A.; Fujii, Kazuyuki; Alabanza, Leah; Ravatn, Roald; Kita, Tsunekazu; Kudoh, Kazuya; Oka, Masahiro; Chin, Khew-Voon

2010-01-01

88

Quantitative Proteomic Analysis of Serum Proteins from Oral Cancer Patients: Comparison of Two Analytical Methods  

PubMed Central

Serum proteomic analysis can be a valuable approach for the discovery of protein biomarkers for early detection or monitoring of a disease. In this study, two analytical methods were compared for quantification of serum proteins in patients with oral cancer. In the first approach, we quantified serum proteins between oral squamous cell carcinoma (OSCC) and healthy control subjects by performing in-solution digestion of serum proteins, isobaric tags for relative and absolute quantification (iTRAQ) labeling of the resulting peptides, strong cation exchange (SCX) fractionation of labeled peptides and finally capillary liquid chromatography with tandem mass spectrometry (LC-MS/MS) analysis of the peptides. In the second approach, we first separated serum proteins with SDS-PAGE. The gel-separated proteins were then digested with trypsin and the resulting peptides were labeled with iTRAQ and analyzed with LC-MS/MS for protein quantification. A total of 319 serum proteins were quantified with the first proteomic approach whereas a total of 281 proteins were quantified by the second proteomic approach. Most of the proteins were identified and quantified by both approaches, suggesting that these methods are similarly effective for serum proteome analysis. This study provides compelling evidence that quantitative serum proteomic analysis of OSCC is a valuable approach for identifying differentially expressed proteins in cancer patients’ circulation systems that may be used as potential biomarkers for disease detection. Further validation in large oral cancer patient populations may lead to a simple and low invasive clinical tool for OSCC diagnosis or monitoring. PMID:25196439

Yang, Yan; Huang, Junwei; Rabii, Bahareh; Rabii, Ramin; Hu, Shen

2014-01-01

89

Serum Microarrays for Large Scale Screening of Protein Levels  

Microsoft Academic Search

There is a great need for comprehensive proteomic anal- ysis of large patient cohorts of plasma and serum samples to identify biomarkers of human diseases. Here we de- scribe a new antibody-based proteomic approach involv- ing a reverse array format where serum samples are spot- ted on a microarray. This enables all samples to be screened for their content of

Magdalena Janzi; Jenny Odling; Qiang Pan-Hammarstrom; Mårten Sundberg; Joakim Lundeberg; Mathias Uhlen; Lennart Hammarstrom; Peter Nilsson

2005-01-01

90

Binding of iralukast to serum proteins and erythrocytes: measurements using ultrafiltration and an erythrocyte partitioning method  

Microsoft Academic Search

The binding of iralukast to plasma (or serum) proteins and to erythrocytes was studied in vitro, at +37°C, using the erythrocyte partitioning method (EPM) and\\/or ultrafiltration (UF) with 14C-labelled iralukast. Iralukast was highly bound in human and animal serum (>99%). Similar bound fraction values were obtained with the two methods: in whole human plasma (or serum) 99.8% (EPM) and 99.9%

Danielle Colussi; Carole Parisot; Gilbert Lefèvre

1999-01-01

91

Association between Duffy blood groups and serum level of the pregnancy zone protein.  

PubMed

In a previous study of psoriatic patients we found increased serum levels of the pregnancy zone protein (PZ) in individuals of blood groups O and Fy(a-). In this study we found an association between the PZ serum level and the Fy(a-) blood group also in young healthy males. Thus the association between the PZ serum level and Duffy blood groups appears to apply to both normal and pathological conditions. PMID:489024

Beckman, L; Bergdahl, K; Cedergren, B; Damber, M G; Lidén, S; von Schoultz, B; Stigbrand, T

1979-01-01

92

Inheritance patterns of enzymes and serum proteins of mallard-black duck hybrids  

USGS Publications Warehouse

From 1974 to 1976, a breeding program was used to produce hybrids of black ducks and mallards for the evaluation of inheritance patterns of serum proteins and serum, liver and muscle enzymes. In addition to the crosses designed to produce hybrids, a series of matings in 1975 and 1976 were designed to evaluate inheritance patterns of a hybrid with either a black duck or mallard. At the F1 level, hybrids were easily distinguished using serum proteins. However, once a hybrid was crossed back to either a mallard or black duck, only 12?23% of the progeny were distinguishable from black ducks or mallards using serum proteins and 23?39% using esterases. Muscle, serum and liver enzymes were similar between the two species.

Morgan, R.P., II; Meritt, D.W.; Block, S.B.; Cole, M.A.; Sulkin, S.T.; Lee, F.B.; Henny, C.J.

1984-01-01

93

Inheritance patterns of enzymes and serum proteins of mallard-black duck hybrids  

USGS Publications Warehouse

From 1974 to 1976, a breeding program was used to produce hybrids of black ducks and mallards for the evaluation of inheritance patterns of serum proteins and serum, liver and muscle enzymes. In addition to the crosses designed to produce hybrids, a series of matings in 1975 and 1976 were designed to evaluate inheritance patterns of a hybrid with either a black duck or mallard. At the F1 level, hybrids were easily distinguished using serum proteins. However, once a hybrid was crossed back to either a mallard or black duck, only 12-23% of the progeny were distinguishable from black ducks or mallards using serum proteins and 23-39% using esterases. Muscle, serum and liver enzymes were similar between the two species.

Morgan, R.P., II; Meritt, D.W.; Block, S.B.; Cole, M.

1984-01-01

94

Immunoprecipitation of Serum Albumin with Protein A-Sepharose: A Biochemistry Laboratory Experiment  

NASA Astrophysics Data System (ADS)

An exercise has been designed and optimized to acquaint students with the simple yet powerful technique of immunoprecipitation. Protein A-Sepharose (PA-S) is used as a solid-phase precipitant to recover bovine serum albumin (BSA, the antigen) recognized by anti-BSA antibody (Ab). The high degree of binding specificity between antigen and antibody is illustrated by recovery of BSA from a complex mixture of proteins obtained from wheat germ and chicken breast. Various controls are included for a thorough data analysis. The solid phase of Ag/Ab/PA-S is recovered by centrifugation, thoroughly washed, and treated to dissociate the BSA antigen. Samples are examined by discontinuous denaturing gel electrophoresis (SDS-PAGE) with Coomassie blue staining. The supernatants, containing proteins that are not precipitated, are also analyzed. Antigenic cross-reactivity, ranging from strong to none, is demonstrated in a second part by using serum albumins from seven different sources. Systems can be set up, shaken, and prepared for electrophoresis in a single lab period with time for laboratory lecture and discussion about antibody structure and function, antibody-based methods in general, and immunoprecipitation in particular.

Bohinski, Robert C.

2000-11-01

95

Serum surfactant proteins-A and -D as biomarkers in idiopathic pulmonary fibrosis  

Microsoft Academic Search

ABSTRACT: Idiopathic pulmonary fibrosis (IPF) has a high mortality rate, and current therapies are only marginally effective. A serum biomarker,that predicts clinical outcome would be useful to stage disease, indicate prognosis and the need for aggressive therapy, and help stratify patients for clinical trials. The goals of this study were to determine whether serum levels of surfactant protein- A (SP-A)

K. E. Greene; T. E. King; Y. Kuroki; B. Bucher-Bartelson; L. S. Newman; H. Nagae; R. J. Mason

2002-01-01

96

Acute phase response of serum amyloid A protein and C reactive protein to the common cold and influenza  

Microsoft Academic Search

C reactive protein (CRP) and serum amyloid A protein (SAA) are sensitive and rapid acute phase reactants, and their measurement for monitoring inflammatory disease and assessing the prognosis in secondary amyloidosis is gaining widespread acceptance. The changes in these proteins in eight subjects suffering from natural colds, 15 subjects with experimentally induced colds (rhinoviruses E1, 3, 9, 14, or 31),

J T Whicher; R E Chambers; J Higginson; L Nashef; P G Higgins

1985-01-01

97

Serum protein identification and quantification of the corona of 5, 15 and 80 nm gold nanoparticles  

NASA Astrophysics Data System (ADS)

When nanoparticles (NP) enter the body they come into contact with body fluids containing proteins which can adsorb to their surface. These proteins may influence the NP interactions with the biological vicinity, eventually determining their biological fate inside the body. Adsorption of the most abundantly binding proteins was studied after an in vitro 24 hr incubation of monodisperse, negatively charged 5, 15 and 80 nm gold spheres (AuNP) in mouse serum by a two-step analysis: proteomic protein identification and quantitative protein biochemistry. The adsorbed proteins were separated from non-adsorbed proteins by centrifugation and gel electrophoresis and identified using a MALDI-TOF-MS-Proteomics-Analyzer. Quantitative analysis of proteins in gel bands by protein densitometry, required the focus on predominantly binding serum proteins. Numerous proteins adsorbed to the AuNP depending on their size, e.g. apolipoproteins or complement C3. The qualitative and quantitative amount of adsorbed proteins differed between 5, 15 and 80 nm AuNP. Band intensities of adsorbed proteins decreased with increasing AuNP sizes based not only on their mass but also on their surface area. Summarizing, the AuNP surface is covered with serum proteins containing transport and immune related proteins among others. Hence, protein binding depends on the size, surface area and curvature of the AuNP.

Schäffler, Martin; Semmler-Behnke, Manuela; Sarioglu, Hakan; Takenaka, Shinji; Wenk, Alexander; Schleh, Carsten; Hauck, Stefanie M.; Johnston, Blair D.; Kreyling, Wolfgang G.

2013-07-01

98

Serum C-Reactive Protein and Procalcitonin Kinetics in Patients Undergoing Elective Total Hip Arthroplasty  

PubMed Central

Background. The sensitivity and the specificity of different methods to detect periprosthetic infection have been questioned. The current study aimed to investigate the kinetics of C-reactive protein (CRP) and procalcitonin (PCT) in patients undergoing uncomplicated elective total hip arthroplasty (THA), to provide a better interpretation of their levels in noninfectious inflammatory reaction. Methods. A total of 51 patients were included. Serum CRP and PCT concentrations were obtained before surgery, on the 1st, 3rd, and 7th postoperative days and after discharge on the 14th and 30th days and at 2 years. Results. Both markers were confirmed to increase after surgery. The serum CRP showed a marked increase on the 3rd postoperative day while the peak of serum PCT was earlier, even if much lower, on the first day. Then, they declined slowly approaching the baseline values by the second postoperative week. PCT mean values never exceed concentrations typically related to bacterial infections. Conclusions. CRP is very sensitive to inflammation. It could be the routine screening test in the follow-up of THA orthopaedic patients, but it should be complemented by PCT when there is the clinical suspicion of periprosthetic infection. PMID:24877114

Battistelli, Sandra; Fortina, Mattia; Carta, Serafino; Guerranti, Roberto; Nobile, Francesco; Ferrata, Paolo

2014-01-01

99

Correlation Between Hypertension, C-Reactive Protein and Serum Uric Acid With Psychological Well-being  

PubMed Central

Background: Multiple population-based human studies have established a strong association between increasing levels of serum C-reactive protein, uric acid and subsequent development of hypertension. Objectives: We aimed to investigate the association between mental well-being with presence of hypertension, hyperuricemia and hs-CRP levels. ?? Patients and Methods: This was a cross sectional study of 801 individuals aged 35-85 years old in Broujerd, Iran, included by randomized cluster sampling. General Health Questionnaire (GHQ-12) for assessing mental health/distress level, MONICA standard questions for evaluating hypertension history, serum hs-CRP and Serum Uric Acid (SUA) were evaluated Data were analyzed by appropriate statistical test such as chi-square, T-test and correlation. Results: One hundred eighty five patients (23.1%) had high distress/minor psychiatric disorders. SUA had significant association with hypertension (r = 0.64, P = 0.034). No significant relation was observed between hs-CRP and hypertension. The correlation between GHQ and hs-CRP was not significant but a weak and negative correlation was found between GHQ and SUA SUA (P = 0.012, r = -0.089). Conclusions: The weak and strong correlation among these parameters indicate that mental wellbeing relays on physical wellness and interact with each other; therefore, controlling hypertension along with uric acid control may effect mental health of any kind of patients. PMID:25237581

Maleki, Ali; Samandari, Saeid; Almeida, Osvaldo; Jafarian Kerman, Scott Reza; Abdolvand, Mahdi; Aliyari, Farshid; Foroughi, Saeid

2014-01-01

100

Human serum protein enhances HIV-1 replication and up-regulates the transcription factor AP-1.  

PubMed

In vitro studies on HIV (HIV-1) replication and neutralization are usually performed in human cell cultures supplemented with FBS instead of human serum (HS). Here we show that in contrast to FBS, addition of increasing amounts of human serum from noninfected donors to the cell culture directly correlates with an increase in HIV-1 replication in vitro. This effect is independent of cell line, virus strain, or batch of pooled human serum used. We found that human serum affects viral transcription in a dose-dependent manner by activating the activator protein-1 (AP-1) member proteins c-FOS, JunD, and JunB in TZM-bl cells. Analysis of the human serum component responsible for this effect indicates that it is a protein having a molecular mass between 250 and 300 kDa. This serum protein, HIV-1 enhancing serum protein (HESP), might promote viral transcription in vivo and consequently play a role in disease progression. PMID:23047699

Perdomo, Maria F; Hosia, Waltteri; Jejcic, Alenka; Corthals, Garry L; Vahlne, Anders

2012-10-23

101

Relief of amplification inhibition in PCR with bovine serum albumin or T4 gene 32 protein  

SciTech Connect

The benefits of adding bovine serum albumin (BSA) or T4 gene 32 proteins (gp32) to PCR were evaluated with reaction mixtures containing substances that inhibit amplification. Whereas 10- to 1,000-fold more FeCl{sub 3}, hemin, fulvic acids, humic acids, tannic acids, or extracts from feces, freshwater, or marine water were accommodated in PCR when either 400 ng of BSA per {mu}l was included in the reactions, neither BSA nor gp32 relieved interference significantly when minimum inhibitory levels of bile salts, bilirubin, EDTA, NaCl, sodium dodecyl sulfate, or Triton X-100 were present. Use of BSA and gp32 together offered no more relief of inhibition than either alone at its optimal level, and neither protein had any noticeable effect on amplification in the absence of inhibitors. 21 refs., 3 figs.

Kreader, C.A. [Environmental Protection Agency, Cincinnati, OH (United States)

1996-03-01

102

Effect of water salinity on total protein and electrophoretic pattern of serum proteins of grass carp, Ctenopharyngodon idella  

PubMed Central

In this study the effects of water salinity on serum total protein and its components in grass carp were investigated. The aim of this study was to determine the effect of salinity tolerance of fish on total serum protein level and its components as an indicator of liver and kidney activity. One hundred and twenty grass carp were divided into four groups, randomly. The first three groups were reared in concentration of 4, 8 and 12 g L-1 of salt solution, respectively, and the fourth group was reared in freshwater and served as control. After 3 weeks, blood samples were collected and after harvesting the blood serum, serum total protein and protein components were measured with Biuret and electrophoresis methods, respectively. Results showed that mean value of serum total protein in the control and three salinities groups were 2.75, 3.28, 2.90 and 3.13 g dL-1, respectively. Five fractions of serum protein were electrophoretically observed as: albumin (Alb), alpha-1 globulin (?1-glu), alpha-2 globulin (?2-glu), beta globulin (?-glu) and gamma globulin (?-glu). There were not any significant differences between the average mean of serum total protein of experimental and control groups (p > 0.05). However, Alb, ?1-glu and ?-glu levels in the experimental groups were significantly higher than in the control group (p < 0.05). The average of ?2-glu and ?-glu revealed no significant difference between the experimental groups (p > 0.05). In conclusion, our results showed that increasing water salinity could have a significant effect on Alb, ?1-glu and ?-glu levels but not on total serum protein in grass carp. PMID:25568723

Peyghan, Rahim; Khadjeh, Gholam Hosain; Enayati, Ala

2014-01-01

103

Pulmonary surfactant proteins and polymer combinations reduce surfactant inhibition by serum.  

PubMed

Acute respiratory distress syndrome (ARDS) is an inflammatory condition that can be associated with capillary leak of serum into alveoli causing inactivation of surfactant. Resistance to inactivation is affected by types and concentrations of surfactant proteins, lipids, and polymers. Our aim was to investigate the effects of different combinations of these three components. A simple lipid mixture (DPPC/POPG) or a more complex lipid mixture (DPPC/POPC/POPG/cholesterol) was used. Native surfactant proteins SP-B and SP-C obtained from pig lung lavage were added either singly or combined at two concentrations. Also, non-ionic polymers polyethylene glycol and dextran and the anionic polymer hyaluronan were added either singly or in pairs with hyaluronan included. Non-ionic polymers work by different mechanisms than anionic polymers, thus the purpose of placing them together in the same surfactant mixture was to evaluate if the combination would show enhanced beneficial effects. The resulting surfactant mixtures were studied in the presence or absence of serum. A modified bubble surfactometer was used to evaluate surface activities. Mixtures that included both SP-B and SP-C plus hyaluronan and either dextran or polyethylene glycol were found to be the most resistant to inhibition by serum. These mixtures, as well as some with either SP-B or SP-C with combined polymers were as or more resistant to inactivation than native surfactant. These results suggest that improved formulations of lung surfactants are possible and may be useful in reducing some types of surfactant inactivation in treating lung injuries. PMID:21741354

Lu, Karen W; Pérez-Gil, Jesús; Echaide, Mercedes; Taeusch, H William

2011-10-01

104

Improved medium for clonal growth of human diploid fibroblasts at low concentrations of serum protein  

Microsoft Academic Search

Summary  A new medium (MCDB 104) has been developed which will support clonal growth of WI-38 cells at concentrations of serum protein\\u000a as low as 25 ?g per ml (equivalent to 0.05% serum). The principal factors responsible for reduction of the protein requirement\\u000a are: (a) adjustment of all nutrient concentrations in medium F12 to experimentally determined optimum values for WI-38 cells;

Wallace L. McKeehan; Kerstin A. McKeehan; Susan L. Hammond; Richard G. Ham

1977-01-01

105

Effect of serum growth factors on intracellular protein degradation in cell culture  

E-print Network

EFFECT OF SERUM GROWTH FACTORS ON INTRACELLULAR PROTEIN DEGRADATION IN CELL CULTURE A Thesis by JOHN BERNARD BODNER Submitted to the Graduate College of Texas A&M University in partial fulfillment of the requirement for the degree of MASTER... OP SCIENCE December 1980 Major Subject: Biochemistry EFFECTS OF SERUM GROWTH FACTORS ON INTRACELLULAR PROTEIN DEGRADATION IN CELL CULTURE A Thesis by JOHN BERNARD BODNER Approved as to style and content by: (', (Chairs(an of C~o' ttee...

Bodner, John Bernard

2012-06-07

106

Smoking and serum proteins in atomic-bomb survivors in Japan  

Microsoft Academic Search

Associations of smoking habit with serum levels of total protein as well as protein fractions were studied in a population consisting of 4,739 atomic-bomb survivors and unexposed control subjects in Hiroshima, Japan who participated in the 1979-1981 period of the Adult Health Study, an ongoing health follow-up program of the Radiation Effects Research Foundation. Smoking was strongly related to serum

D. O. Stram; S. Akiba; K. Neriishi; R. G. Stevens; Y. Hosoda

1990-01-01

107

A quantitative proteomic approach to identify significantly altered protein networks in the serum of patients with lymphangioleiomyomatosis (LAM).  

PubMed

Lymphangioleiomyomatosis (LAM) is a rare and progressive cystic lung condition affecting approximately 3.4-7.5/million women, with an average lag time between symptom onset and diagnosis of upwards of 4 years. The aim of this work was to identify altered proteins in LAM serum which may be potential biomarkers of disease. Serum from LAM patient volunteers and healthy control volunteers were pooled and analysis carried out using quantitative 4-plex iTRAQ technology. Differentially expressed proteins were validated using ELISAs and pathway analysis was carried out using Ingenuity Pathway Analysis. Fourteen proteins were differentially expressed in LAM serum compared to control serum (p<0.05). Further screening validated the observed differences in extracellular matrix remodelling proteins including fibronectin (30% decrease in LAM, p?=?0.03), von Willebrand Factor (40% reduction in LAM, p?=?0.03) and Kallikrein III (25% increase in LAM, p?=?0.03). Pathway networks elucidated the relationships between the ECM and cell trafficking in LAM. This study was the first to highlight an imbalance in networks important for remodelling in LAM, providing a set of novel potential biomarkers. These understandings may lead to a new effective treatment for LAM in the future. PMID:25133674

Banville, Nessa; Burgess, Janette K; Jaffar, Jade; Tjin, Gavin; Richeldi, Luca; Cerri, Stefania; Persiani, Elisa; Black, Judith L; Oliver, Brian G

2014-01-01

108

Serum levels of rapid turnover proteins are decreased and related to systemic inflammation in patients with ovarian cancer  

PubMed Central

Poor nutritional status is common in ovarian cancer. It is well known that the nutritional status of a patient with malignant disease is associated with survival, and that it can be assessed by serum levels of rapid turnover proteins (RTPs), such as retinol binding protein, prealbumin and transferrin. Systemic inflammation, usually observed in the form of elevated C-reactive protein (CRP) or neutrophil/lymphocyte ratio (NLR), occurs by various mechanisms involving numerous pro-inflammatory cytokines. These include interleukin (IL)-17 and other soluble protein mediators, such as soluble IL-2 receptor (sIL-2R) and vascular endothelial growth factor (VEGF). In this study, circulating levels of RTP were decreased in advanced stages of ovarian cancer, and significant inverse correlations were found between RTP levels and serum levels of CRP or NLR. CRP levels were also correlated with serum levels of VEGF and sIL-2R. Moreover, NLR, VEGF and sIL-2R levels, and IL-17 production, were all inversely correlated with RTP levels. These findings indicate that chronic inflammation may be associated with compromised immune function, such as an impaired T-cell response, via various inflammatory proteins, including sIL-2R, VEGF and IL-17. The key mechanisms leading to cancer cachexia, in which nutritional impairment is a major clinical issue, appear to be primarily immune reactions caused by chronic inflammation. Anti-inflammatory treatments may be effective in clinically improving various symptoms associated with these mechanisms. PMID:24396450

WATANABE, TAKAFUMI; SHIBATA, MASAHIKO; NISHIYAMA, HIROSHI; SOEDA, SHU; FURUKAWA, SHIGENORI; GONDA, KENJI; TAKENOSHITA, SEIICHI; FUJIMORI, KEIYA

2014-01-01

109

Serum levels of club (Clara) cell secretory protein predict cancer mortality in adults  

PubMed Central

Background Club (formerly Clara) cell secretory protein (CC16) is produced mainly by bronchiolar club cells and has been shown to have protective effects against airway inflammation and oxidative stress from cigarette smoking and related carcinogens. The goal of this study was to determine whether serum CC16 levels predict all-cause and cancer-specific mortality in adults. Methods We used data from the population-based TESAOD study, a prospective cohort study of respiratory health initiated in Tucson, AZ in 1972. At baseline, participants completed standardized respiratory questionnaires and lung function tests. Serum CC16 was measured in cryopreserved serum samples. A review of vital status of participants as of January 1st, 2011 was completed through contact with next of kin, collection of death certificates, and linkage with the National Death Index. Findings A total of 1086 participants who were 21 to 70 years old at enrollment were included. Of these, 653 (60%) died by 2011 and cause of death was ascertained for 649 (99%). In Cox proportional hazards models adjusted for sex, age, education, body mass index categories, smoking and pack-years, and baseline levels of lung function, serum CC16 levels at the baseline survey were inversely associated with mortality risk over the study follow-up. Mortality risk increased by 16% for each standard deviation (SD) decrease in CC16 (Hazard Ratio (HR), 95% CI: 1.16, 1.06 – 1.26; p = 0.0007). When data on cause-specific mortality were analyzed, each SD decrease in serum CC16 was associated with >40% increased risk of dying of cancer (adjusted HR=1.41, 1.19 – 1.67; p < 0.0001). Among smokers, the corresponding adjusted HRs for mortality by lung cancer were 1.52 (1.14 – 2.03; p = 0.004). Interpretation Serum CC16 levels predict mortality risk in the general adult population. The excess risk associated with lower CC16 is largely explained by cancer, particularly lung cancer. PMID:24461757

Guerra, Stefano; Vasquez, Monica M.; Spangenberg, Amber; Halonen, Marilyn; Martinez, Fernando D.

2014-01-01

110

Serum protein electrophoresis by using high-resolution agarose gel in clinically healthy and Aspergillus species-infected falcons.  

PubMed

Serum protein electrophoresis has gained importance in avian medicine during the past decade. Interpretation of electrophoretic patterns should be based on species-specific reference intervals and the electrophoresis gel system. In this study, serum protein electrophoresis by using high-resolution agarose gels was performed on blood samples collected from 105 falcons, including peregrine falcons (Falco peregrinus), gyrfalcons (Falco rusticolus), saker falcons (Falco cherrug), red-naped shaheens (Falco pelegrinoides babylonicus), and hybrid falcons, that were submitted to the Dubai Falcon Hospital (Dubai, United Arab Emirates) between 2003 and 2006. Reference values were established in clinically healthy birds and compared with values from falcons infected with Aspergillus species (n = 32). Falcons with confirmed aspergillosis showed significantly lower prealbumin values, which is a novel finding. Prealbumin has been documented in many avian species, but further investigation is required to illuminate the diagnostic significance of this negative acute-phase protein. PMID:23409432

Kummrow, Maya; Silvanose, Christudas; Di Somma, Antonio; Bailey, Thomas A; Vorbrüggen, Susanne

2012-12-01

111

Serum Golgi protein 73 levels and liver pathological grading in cases of chronic hepatitis B.  

PubMed

The present study was designed to assess the correlation between serum Golgi protein 73 (GP73) and liver pathological grading and staging in patients with chronic hepatitis B (CHB). Two hundred and fifty?three patients with chronic hepatitis B virus (HBV) infections were enrolled in the present study. All patients received a serum GP73 test, and 91 CHB patients underwent liver biopsy. GP73 expression in liver tissue was assessed by immunohistochemical analysis. The results indicated that serum GP73 levels were positively correlated with disease progression in patients with chronic HBV infection (r=0.677). There was no significant difference in serum GP73 levels between hepatitis B e antigen?positive and ?negative patients (P>0.05). There were also no significant differences in serum GP73 levels among specimens with varying HBV DNA contents (P>0.05). Serum GP73 levels were positively correlated with increased liver pathological grading (r=0.737) and staging (r=0.692), and immunohistochemical analysis indicated that GP73 protein expression increased concurrently with liver pathological grading and staging. In conclusion, serum GP73 was found to be correlated with liver pathological grading and staging in patients with CHB, and may be an effective indicator for the evaluation of disease progression. However, serum GP73 levels were not associated with HBV replication. PMID:25524053

Xu, Zhengju; Pan, Xingnan; Wei, Kaipeng; Ding, Hongbing; Wei, Meijuan; Yang, Huanwen; Liu, Qian

2015-04-01

112

The role of the female Syrian hamster protein on the interaction between serum lipoproteins and heparin.  

PubMed

We have previously shown the effect of phosphorylcholine-binding proteins from rat (PCBP) and rabbit (CRP) on the precipitation of serum lipoproteins by heparin in presence of Ca2+. The present paper describes the effect of a phosphorylcholine-binding protein from the female Syrian hamster (FP) on the lipoprotein precipitation reaction. The precipitation of lipoproteins by heparin was lower in assays using female hamster serum in which FP is a prominent protein, compared with assays with male serum in which FP is present in very low concentration. Depletion of FP from female serum resulted in increased lipoprotein precipitation. The addition of purified FP to assays using human very low density lipoprotein (VLDL) inhibited the precipitation reaction. The precipitation of lipoproteins was also examined using serum from male hamsters treated with diethylstilbestrol and female hamsters treated with testosterone, treatments known to modulate the levels of FP. Results indicate an inverse relationship between serum FP levels from normal and hormone-treated hamsters and the precipitation of lipoproteins from their serum. The partially desialylated FP when added to precipitation assays using human VLDL resulted in reduced inhibition of VLDL precipitation. PMID:3620160

Saxena, U; Nagpurkar, A; Coe, J E; Mookerjea, S

1987-05-01

113

Differential phosphorylation of paxillin in response to surface-bound serum proteins during early osteoblast adhesion.  

PubMed

An early signaling event during the adhesion and spreading of cells is integrin-mediated tyrosine phosphorylation of the cytoskeletal adaptor protein paxillin and the non-receptor tyrosine kinase pp125(FAK) at focal contacts. To determine the influence of surface-charge and -adsorbed adhesion proteins on this signaling pathway, paxillin phosphorylation was examined during attachment of MC3T3-E1 osteoblast-like cell onto charged and uncharged polystyrene, and on adsorbed layers of serum proteins, fibronectin (Fn), vitronectin (Vn), a mixture of Fn and Vn, and albumin. Paxillin phosphorylation was induced 2.4-fold (P < 0.05) on charged vs uncharged polystyrene only in the presence of serum proteins. Activation of paxillin via Fn or Vn alone, or in combination, resulted in significantly lower phosphorylation signals compared to whole serum (41 +/- 6.9%, P < 0.05, 45 +/- 5.9%, P < 0.05, and 76 +/- 9.8%, P < 0.075, respectively). Confocal laser microscopy confirmed increased co-localization of phosphotyrosine and paxillin at protruding lamellopodia of spreading osteoblasts on charged vs uncharged serum-pretreated polystyrene. Taken together, these data suggest that subtle differences in surface characteristics mediate effects on adhering cells via adsorbed serum proteins involving the cytoskeletal adaptor protein paxillin. PMID:11444850

Sommerfeldt, D W; McLeod, K J; Rubin, C T; Hadjiargyrou, M

2001-07-13

114

A panel of regulated proteins in serum from patients with cervical intraepithelial neoplasia and cervical cancer.  

PubMed

We developed a discovery-validation mass-spectrometry-based pipeline to identify a set of proteins that are regulated in serum of patients with cervical intraepithelial neoplasia (CIN) and squamous cell cervical cancer using iTRAQ, label-free shotgun, and targeted mass-spectrometric quantification. In the discovery stage we used a "pooling" strategy for the comparative analysis of immunodepleted serum and revealed 15 up- and 26 down-regulated proteins in patients with early- (CES) and late-stage (CLS) cervical cancer. The analysis of nondepleted serum samples from patients with CIN, CES, an CLS and healthy controls showed significant changes in abundance of alpha-1-acid glycoprotein 1, alpha-1-antitrypsin, serotransferrin, haptoglobin, alpha-2-HS-glycoprotein, and vitamin D-binding protein. We validated our findings using a fast UHPLC/MRM method in an independent set of serum samples from patients with cervical cancer or CIN and healthy controls as well as serum samples from patients with ovarian cancer (more than 400 samples in total). The panel of six proteins showed 67% sensitivity and 88% specificity for discrimination of patients with CIN from healthy controls, a stage of the disease where current protein-based biomarkers, for example, squamous cell carcinoma antigen (SCCA), fail to show any discrimination. Additionally, combining the six-protein panel with SCCA improves the discrimination of patients with CES and CLS from healthy controls. PMID:25232869

Boichenko, Alexander P; Govorukhina, Natalia; Klip, Harry G; van der Zee, A G J; Güzel, Co?kun; Luider, Theo M; Bischoff, Rainer

2014-11-01

115

Relationship between Acute Phase Proteins and Serum Fatty Acid Composition in Morbidly Obese Patients  

PubMed Central

Background. Obesity is considered a low-grade inflammatory state and has been associated with increased acute phase proteins as well as changes in serum fatty acids. Few studies have assessed associations between acute phase proteins and serum fatty acids in morbidly obese patients. Objective. To investigate the relationship between acute phase proteins (C-Reactive Protein, Orosomucoid, and Albumin) and serum fatty acids in morbidly obese patients. Methods. Twenty-two morbidly obese patients were enrolled in this study. Biochemical and clinical data were obtained before bariatric surgery, and fatty acids measured in preoperative serum. Results. Orosomucoid was negatively correlated with lauric acid (P = 0.027) and eicosapentaenoic acid (EPA) (P = 0.037) and positively with arachidonic acid (AA) (P = 0.035), AA/EPA ratio (P = 0.005), and n-6/n-3 polyunsaturated fatty acids ratio (P = 0.035). C-Reactive Protein (CRP) was negatively correlated with lauric acid (P = 0.048), and both CRP and CRP/Albumin ratio were negatively correlated with margaric acid (P = 0.010, P = 0.008, resp.). Albumin was positively correlated with EPA (P = 0.027) and margaric acid (P = 0.008). Other correlations were not statistically significant. Conclusion. Our findings suggest that serum fatty acids are linked to acute phase proteins in morbidly obese patients. PMID:24167354

Fernandes, Ricardo; Beserra, Bruna Teles Soares; Cunha, Raphael Salles Granato; Hillesheim, Elaine; Camargo, Carolina de Quadros; Pequito, Danielle Cristina Tonello; de Castro, Isabela Coelho; Fernandes, Luiz Cláudio; Nunes, Everson Araújo; Trindade, Erasmo Benício Santos de Moraes

2013-01-01

116

Evaluation of IGFBP-7 DNA methylation changes and serum protein variation in Swedish subjects with and without type 2 diabetes  

PubMed Central

Background Insulin-like growth factor-binding protein 7 (IGFBP-7) is able to interact with insulin-like growth factor 1 (IGF-1) as well as insulin. Previous studies have suggested that serum IGFBP-7 levels may be associated with insulin resistance in type 2 diabetes (T2D). This study aimed to evaluate IGFBP-7 serum protein and IGFBP7 DNA methylation levels in the subjects with and without T2D. Results A total of 340 Swedish subjects including 100 newly diagnosed T2D patients (50 women/50 men), 100 age-matched nondiabetic control subjects (50/50) and 140 treated T2D patients (54/86) were studied. Serum IGFBP-7 levels were measured with a novel ELISA. IGF1, IGFBP-1, and insulin were determined by in-house radioimmunoassays. DNA methylation levels in the IGFBP7 gene were analyzed with a bisulfite-pyrosequencing technique. Serum IGFBP-7 protein levels were similar among nondiabetic subjects, newly diagnosed, and treated T2D patients and were not correlated with IGFBP7 DNA methylation. However, IGFBP7 DNA methylation was increased in men with newly diagnosed T2D compared with nondiabetic controls (17.6% vs. 12.5%, P < 0.01). Serum IGFBP-7 levels correlated (r = 0.331, P = 0.019) with serum IGFBP-1 levels, a marker of insulin production, in men but not women with newly diagnosed T2D. Conclusions This study demonstrates for the first time that IGFBP7 DNA methylation levels are increased in Swedish men with newly diagnosed T2D. The correlation between IGFBP-7 and IGFBP-1 suggests that low IGFBP-7 may be associated with insulin resistance in T2D. PMID:24180466

2013-01-01

117

Cockroach Larval-specific Protein, a Tyrosine-rich Serum Protein* (Received for publication, June9, 1983)  

E-print Network

Cockroach Larval-specific Protein, a Tyrosine-rich Serum Protein* (Received for publication, June9 in the hemolymph of cockroaches shortly be- fore molting, but is rapidly cleared from the hemo- lymph during themolting cycle of cockroaches has been studied in syn- chronouslymoltingcultures (1-3). Among

Kunkel, Joseph G.

118

Serum Vascular Adhesion Protein-1 Predicts 10-Year Cardiovascular and Cancer Mortality in Individuals With Type 2 Diabetes  

PubMed Central

OBJECTIVE Vascular adhesion protein-1 (VAP-1) participates in inflammation and catalyzes the breakdown of amines to produce aldehyde, hydrogen peroxide, and ammonia. Serum VAP-1 correlates positively with both acute hyperglycemia and diabetes. We conducted a cohort study to evaluate whether serum VAP-1 predicts 10-year survival in type 2 diabetic patients. RESEARCH DESIGN AND METHODS Between July 1996 and June 2003, we enrolled 661 type 2 diabetic subjects at National Taiwan University Hospital. Serum VAP-1 in the samples obtained at enrollment was measured by time-resolved immunofluorometric assay. The vital status of all subjects was ascertained by linking their data with computerized death certificates in Taiwan. RESULTS The medium follow-up period was 10.4 years. Subjects with serum VAP-1 in the highest tertile had a hazard ratio (HR) of 2.19 (95% CI 1.17–4.11) for all-cause mortality adjusted for age, sex, smoking, history of cardiovascular disease, obesity, hypertension, hemoglobin A1c, diabetes duration, total cholesterol, use of statins, abnormal ankle-brachial index, estimated glomerular filtration rate (eGFR), and proteinuria. The adjusted HRs for logarithmically transformed serum VAP-1 were 5.83 (95% CI 1.17–28.97) for cardiovascular mortality, 6.32 (95% CI 1.25–32.00) for mortality from cardiovascular and diabetic causes, and 17.24 (95% CI 4.57–65.07) for cancer mortality. There were four variables, including age, serum VAP-1, proteinuria, and eGFR, which could enhance mortality prediction significantly. CONCLUSIONS Serum VAP-1 can predict 10-year all-cause mortality, cardiovascular mortality, and cancer mortality independently in type 2 diabetic subjects. Serum VAP-1 is a novel biomarker that improves risk prediction over and above established risk factors. PMID:21282368

Li, Hung-Yuan; Jiang, Yi-Der; Chang, Tien-Jyun; Wei, Jung-Nan; Lin, Mao-Shin; Lin, Cheng-Hsin; Chiang, Fu-Tien; Shih, Shyang-Rong; Hung, Chi Sheng; Hua, Cyue-Huei; Smith, David J.; Vanio, Jani; Chuang, Lee-Ming

2011-01-01

119

Detection of   and   Light Chain Monoclonal Proteins in Human Serum: Automated Immunoassay versus Immunofixation Electrophoresis  

Microsoft Academic Search

Recently, turbidimetric immunoassays for detecting and quantifying and free light chains (FLC) have become available and are promoted as being more sensitive than immunofixation electrophoresis (IFE) in detecting FLC monoclonal proteins. In this study, we assessed the ability of these turbidimetric assays to detect serum monoclonal proteins involving both free and heavy-chain-bound and light chains compared to standard immunofixation electrophoresis.

Troy D. Jaskowski; Christine M. Litwin; Harry R. Hill

2006-01-01

120

ON THE INFLUENCE OF IRRADIATION ON THE COMPOSITION OF SERUM PROTEINS  

Microsoft Academic Search

Rabbits were irradiated with 700 r and serum protein fractions were ; examined electrophoretically 11 and 14 days later. The most marked changes in ; protein fractions were noted on the 11th day, at which time the relative ; percentage of albwnin had fallen to 44 compared with 61% in nonirradiated ; controls; gamma -globulin had fallen to 4 from

E. Waldschmidt-Leitz; L. Keller

1961-01-01

121

Tissue localization and chromosomal assignment of a serum protein that tracks the cystic fibrosis gene  

Microsoft Academic Search

The basic gene defect in the autosomal recessive disorder cystic fibrosis has not been identified, and no firm linkage of the disorder to any other marker has been reported. However, a serum protein abnormality present in unaffected heterozygotes as well as in affected homozygotes has been described1, and immunological quantitation of this protein, termed cystic fibrosis antigen, allows the three

Veronica van Heyningen; Caroline Hayward; Judy Fletcher; Christine McAuley

1985-01-01

122

The use of proteomics in identifying differentially expressed serum proteins in humans with type 2 diabetes  

PubMed Central

Background The aim of the study was to optimize protocols for finding and identifying serum proteins that are differentially expressed in persons with normal glucose tolerance (NGT) compared to individuals with type 2 diabetes mellitus (T2DM). Serum from persons with NGT and persons with T2DM was profiled using ProteinChip arrays and time-of-flight mass spectra were generated by surface enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS). Results Mass spectra from NGT- and T2DM-groups were compared. Fifteen proteins ranging from 5 to 79 kDa were differentially expressed (p < 0.05). Five of these proteins showed decreased and ten showed increased serum levels in individuals with T2DM. To be able to identify the proteins, the complexity of the sample was reduced by fractionation approaches. Subsequently, the purified fractions containing biomarkers were separated by one-dimensional SDS-polyacrylamide gel electrophoresis (SDS-PAGE) in two identical lanes. Protein bands of the first lane were excised and subjected to passive elution to recapture the biomarkers on ProteinChip arrays. The corresponding bands of the second lane were subjected to peptide-mass fingerprinting (PMF). Using this approach four of the differentially expressed proteins were identified as apolipoprotein C3 (9.4 kDa), transthyretin (13.9 kDa), albumin (66 kDa) and transferrin (79 kDa). Whereas apolipoprotein C3 and transthyretin were up-regulated, albumin and transferrin were down-regulated in T2DM. Conclusion Protocols for protein profiling by SELDI-TOF MS and protein identification by fractionation, SDS-PAGE and PMF were optimized for serum from humans with T2DM. With these protocols differentially expressed proteins were discovered and identified when serum from NGT- and T2DM-individuals was analyzed. PMID:17163994

Sundsten, Tea; Eberhardson, Michael; Göransson, Michael; Bergsten, Peter

2006-01-01

123

Effects of serum proteins on intracellular uptake and cytotoxicity of carbon nanoparticles  

Microsoft Academic Search

To explore the effects of the novel properties of carbon nanoparticles (CNPs) on cytotoxicity, the adsorption of serum proteins in cell culture medium on multi-walled carbon nanotubes and three kinds of carbon blacks was investigated. The uptake of CNPs by Hela cells was measured quantitatively using 99mTc radionuclide labeling and tracing techniques, and the dependence of CNPs uptake on serum

Ying Zhu; Wenxin Li; Qingnuan Li; Yuguo Li; Yufeng Li; Xiaoyong Zhang; Qing Huang

2009-01-01

124

Serum proteins in chronic hepatitis B patients treated with peginterferon alfa-2b  

PubMed Central

AIM: To study the differential protein profile in serum of hepatitis B patients. METHODS: Serum samples were obtained from patients with chronic hepatitis B who were receiving peginterferon alfa-2b. The serum samples were subjected to albumin depletion and analyzed by two-dimensional gel electrophoresis (2-DE). Differentially expressed protein spots were identified by electrospray ionization-quadrupole time-of-flight mass spectrometry. Alpha-2-HS-glycoprotein, complement component C3c and CD5 antigen were further analyzed by an enzyme-linked immunosorbent assay and immunonephelometry. RESULTS: Nineteen patients with HBeAg-positive chronic hepatitis B (CHB) were studied. These patients were followed for at least 1 year after treatment and were classified according to their treatment response: responders (n = 9) and non-responders (n = 10). 2-DE and MS/MS analysis were performed to compare the serum proteins before initiating peginterferon alfa-2b. From the quantitative analysis of the 2-D gel, 7 proteins were detected between the two groups at different levels before treatment. Among these potential candidates, serum levels of alpha-2-HS-glycoprotein, complement component C3c and CD5 antigen-like precursor were further analyzed. In the validation phase, 23 subjects, 9 sustained responders and 14 non-responders, were recruited. Interestingly, the levels of alpha-2-HS-glycoprotein and complement component C3c were elevated in the serum of the non-responders compared to the responders. CONCLUSION: Serum alpha-2-HS-glycoprotein and complement component C3c may be potential serum biomarkers in predicting the treatment response of peginterferon alfa-2b in patients with CHB prior to treatment. PMID:23964140

Kuakarn, Sunida; SomParn, Poorichaya; Tangkijvanich, Pisit; Mahachai, Varocha; Thongboonkerd, Visith; Hirankarn, Nattiya

2013-01-01

125

Probing thyroglobulin in undiluted human serum based on pattern recognition and competitive adsorption of proteins  

NASA Astrophysics Data System (ADS)

Thyroglobulin (Tg) is a sensitive indicator of persistent or recurrent differentiated thyroid cancer of follicular cell origin. Detection of Tg in human serum is challenging as bio-receptors, such as anti-Tg, used in immunoassay have relatively weak binding affinity. We engineer sensing surfaces using the competitive adsorption of proteins, termed the Vroman Effect. Coupled with Surface Plasmon Resonance, the "cross-responsive" interactions of Tg on the engineered surfaces produce uniquely distinguishable multiple signature patterns, which are discriminated using Linear Discriminant Analysis. Tg-spiked samples, down to 2 ng/ml Tg in undiluted human serum, are sensitively and selectively discriminated from the control (undiluted human serum).

Wang, Ran; Huang, Shuai; Li, Jing; Chae, Junseok

2014-10-01

126

Eimeria Species and Genetic Background Influence the Serum Protein Profile of Broilers with Coccidiosis  

PubMed Central

Background Coccidiosis is an intestinal disease caused by protozoal parasites of the genus Eimeria. Despite the advent of anti-coccidial drugs and vaccines, the disease continues to result in substantial annual economic losses to the poultry industry. There is still much unknown about the host response to infection and to date there are no reports of protein profiles in the blood of Eimeria-infected animals. The objective of this study was to evaluate the serum proteome of two genetic lines of broiler chickens after infection with one of three species of Eimeria. Methodology/Principal Findings Birds from lines A and B were either not infected or inoculated with sporulated oocysts from one of the three Eimeria strains at 15 d post-hatch. At 21 d (6 d post-infection), whole blood was collected and lesion scoring was performed. Serum was harvested and used for 2-dimensional gel electrophoresis. A total of 1,266 spots were quantitatively assessed by densitometry. Protein spots showing a significant effect of coccidia strain and/or broiler genetic line on density at P<0.05?0.01 (250 spots), P<0.01?0.001 (248 spots), and P<0.001 (314 spots) were excised and analyzed by matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry. Proteins were identified in 172 spots. A total of 46 different proteins were identified. Of the spots with a corresponding protein identification, 57 showed a main effect of coccidia infection and/or 2-way interaction of coccidia infection×broiler genetic line at P<0.001. Conclusions/Significance Several of the metabolic enzymes identified in this study are potential candidates for early diagnostic markers of E. acervulina infection including malate dehydrogenase 2, NADH dehydrogenase 1 alpha subcomplex 9, and an ATP synthase. These proteins were detected only in Line A birds that were inoculated with E. acervulina. Results from this study provide a basic framework for future research aimed at uncovering the complex biochemical mechanisms involved in host response to Eimeria infection and in identifying molecular targets for diagnostic screening and development of alternative preventative and therapeutic methods. PMID:21297942

Gilbert, Elizabeth R.; Cox, Chasity M.; Williams, Patricia M.; McElroy, Audrey P.; Dalloul, Rami A.; Ray, W. Keith; Barri, Adriana; Emmerson, Derek A.; Wong, Eric A.; Webb, Kenneth E.

2011-01-01

127

Tubule urate and PAH transport: sensitivity and specificity of serum protein inhibition  

SciTech Connect

Macromolecules in rabbit serum inhibit the cellular uptake and transepithelial secretion of (/sup 14/C)urate and p-(/sup 3/H)aminohippurate ((/sup 3/H)PAH) in rabbit S/sub 2/ proximal tubule segments. To understand better the potential role these inhibitors may have in the regulation of renal organic anion excretion, the authors examined the specificity and relative inhibitory effects on tubule urate and PAH transport of albumin and ..gamma..-globulin, the major inhibitory proteins in rabbit serum. Native rabbit serum markedly inhibited the cellular accumulation or urate and PAH by isolated nonperfused segments. Urate and PAH transport was also inhibited by bovine serum, human serum, Cohn-fractionated rabbit albumin, and rabbit ..gamma..-globulin, but not by Cohn-fractionated bovine serum albumin. ..cap alpha..-Lactalbumin and ..beta..-lactoglobulin, derived from milk, also inhibited urate and PAH transport, but to a lesser extent than albumin and ..gamma..-globulin. The transport inhibitory effects of proteins were independent of their binding to urate and PAH. Unidirectional influx and the steady-state intracellular accumulation of urate and PAH in suspensions of proximal tubules were decreased by rabbit serum proteins, suggesting that these inhibitors act on the external face of the cells to diminish the uptake of the organic anions. These studies indicate that the principal plasma proteins (albumin and ..gamma..-globulin) significantly inhibit urate and PAH transporters in the basolateral membranes of S/sub 2/ proximal tubules. They suggest that circulating plasma proteins that can penetrate the basement membrane of proximal tubules may directly modulate the renal excretion of urate and PAH.

Grantham, J.J.; Kennedy, J.; Cowley, B.

1987-04-01

128

Paxillin suppresses the proliferation of HPS rat serum treated PASMCs by up-regulating the expression of cytoskeletal proteins.  

PubMed

Hepatopulmonary syndrome (HPS) is a triad of advanced liver disease, intrapulmonary vasodilatation (IPVD), and arterial hypoxemia. The arterial hypoxemia induces pulmonary vascular remodelling (PVR). In recent studies, the role of the proliferation of pulmonary artery smooth muscle cells (PASMCs) in PVR associated with HPS has been established; the changes in cytoskeletal proteins play an essential role in the proliferation of PASMCs. Little is known about the relevance of cytoskeletal protein expression or the molecular mechanisms of PVR associated with HPS. In addition, it has been identified that paxillin could influence the cytoskeletal protein expression by some important signaling pathways in many diseases, including lung cancer and liver cancer. In this study, we found that HPS rat serum from a common bile duct ligation (CBDL) rat model decreased the expression of cytoskeletal proteins (?-actin, ?-tubulin, and destrin) and enhanced the expression levels of paxillin mRNA and protein in PASMCs. After silencing paxillin with siRNA, we found that the down-regulation of cytoskeletal protein expression, induced by the HPS rat serum, was reversed. Additionally, we reported that HPS rat serum improved the proliferation of PASMCs and down-regulation of paxillin could significantly inhibit this variation. These findings suggest that the up-regulation of cytoskeletal protein expression, induced by the paxillin, may cause the dysregulation of PASMC proliferation as well as play a fundamental role in PVR associated with HPS. In conclusion, down-regulation of paxillin by siRNA results in the inhibition of the dysregulation of cytoskeletal proteins and proliferation of PASMCs, suggesting a potential therapeutic effect on PVR associated with HPS. PMID:24457422

Chen, Yang; Yi, Bin; Wang, Zhi; Gu, Jianteng; Li, Yongshuai; Cui, Jian; Lu, Kaizhi

2014-04-01

129

Prognostic role of serum C-reactive protein in esophageal cancer: a systematic review and meta-analysis  

PubMed Central

Background Recent studies have shown that C-reactive protein (CRP) is a useful predictive factor in several cancers; however, its role in esophageal cancer (EC) is controversial. Methods A systematic literature search was performed using Medline, PubMed, and Web of Science to analyze the prognostic value of serum CRP in patients with EC. A meta-analysis was performed to assess the association between serum CRP and overall survival (OS) in patients with EC. Results A total of eight studies involving 1,471 patients were included in our study. Our pooled results demonstrated that a high level of serum CRP was associated with poor OS (hazard ratio [HR]: 1.40, 95% confidence interval [CI]: 1.25–1.57, I2=81.3%, P<0.0001). Subgroup analyses were performed in further investigations. When the patients were segregated according to treatment, pathological type, and cut-off level, high levels of serum CRP were found to be significantly correlated with OS. Conclusion Our meta-analysis revealed that high levels of serum CRP were significantly associated with poor OS in patients with EC. PMID:25653533

Huang, Ying; Feng, Ji-Feng; Liu, Jin-Shi; Chen, Qi-Xun

2015-01-01

130

Serum-Based Protein Biomarkers in Blast-Induced Traumatic Brain Injury Spectrum Disorder  

PubMed Central

The biological consequences of exposure to explosive blast are extremely complex. Serum protein biomarkers in blast-induced traumatic brain injury (bTBI) can aid in determining injury severity, monitoring progress, and predicting outcome. Exposure to blast results in varying degrees of physical injury. Explosive blast can also induce psychological stress that can contribute to or amplify the extent of physical damage. Given the complexity, scale of injury, and variety of symptoms, bTBI may be best described as a spectrum disorder. In this focused review, we summarize the status of serum protein biomarkers in bTBI in the context of the classification and pathological changes of other forms of TBI. Finally, we recommend specific and easily implementable measures to accelerate serum protein biomarker discovery and validation in bTBI. PMID:22783223

Agoston, Denes V.; Elsayed, Mohammad

2012-01-01

131

Disparate Proteome Responses of Pathogenic and Non-pathogenic Aspergilli to Human Serum Measured by Activity-Based Protein Profiling (ABPP)  

SciTech Connect

Aspergillus fumigatus is the primary pathogen causing the devastating pulmonary disease Invasive Aspergillosis in immunocompromised individuals. Genomic analysis shows high synteny between A. fumigatus and closely related rarely pathogenic Neosartorya fischeri and Aspergillus clavatus genomes. To investigate the presence of unique or highly inducible protein reactivity in the pathogen, we applied activity-based protein profiling to compare protein reactivity of all three fungi over time in minimal media growth and in response to human serum. We found 350 probe-reactive proteins exclusive to A. fumigatus, including known virulence associated proteins, and 13 proteins associated with stress response exclusive to A. fumigatus culture in serum. Though the fungi are highly orthologous, A. fumigatus has significantly more activity across varied biological process. Only 50% of expected orthologs of measured A. fumigatus reactive proteins were observed in N. fischeri and A. clavatus. Human serum induced processes uniquely or significantly represented in A. fumigatus include actin organization and assembly, transport, and fatty acid, cell membrane, and cell wall synthesis. Additionally, signaling proteins regulating vegetative growth, conidiation, and cell wall integrity, required for appropriate cellular response to external stimuli, had higher reactivity over time in A. fumigatus and N. fisheri, but not in A. clavatus. Together, we show that measured proteins and physiological processes identified solely or significantly over-represented in A. fumigatus reveal a unique adaptive response to human protein not found in closely related, but rarely aspergilli. These unique protein reactivity responses may reveal how A. fumigatus initiates pulmonary invasion leading to Invasive Aspergillosis.

Wiedner, Susan D.; Ansong, Charles; Webb-Robertson, Bobbie-Jo M.; Pederson, Leeanna M.; Fortuin, Suereta; Hofstad, Beth A.; Shukla, Anil K.; Panisko, Ellen A.; Smith, Richard D.; Wright, Aaron T.

2013-07-01

132

Suppression of severe lesions, myonecrosis and hemorrhage, caused by Protobothrops flavoviridis venom with its serum proteins.  

PubMed

Protobothrops flavoviridis serum proteins precipitated with ammonium sulfate were chromatographed on a DEAE-Toyopearl 650M column at pH 7.5 with stepwise increase or with linear gradient of NaCl concentration. Peaks 3 and 4 serum proteins, obtained by linear gradient elution and named Fr(de3) and Fr(de4), contained Habu serum factors (HSF) and phospholipase A2 (PLA2) inhibitors (PfPLI), respectively. The serum proteins eluted at 0.2 M NaCl by stepwise elution, named Fr(0.2NaCl), effectively suppressed myonecrosis and hemorrhage caused by P. flavoviridis venom in rat or mouse thigh muscles. The Fr(0.2NaCl) were fractionated by HPLC and the fractions, after SDS-PAGE, underwent far-western blot analysis with PLA2 ([Asp(49)]PLA2) and BPI ([Lys(49)]PLA2) as the probes. Four PfPLIs, namely, Pf?PLI-A, Pf?PLI-B, Pf?PLI-A and Pf?PLI-B, were identified together with their selective binding specificities to PLA2 species. In addition, a new 9 kDa protein, which is specifically bound to BPI, was found. Suppression of P. flavoviridis venom-induced severe lesions, such as myonecrosis, hemorrhage and edema, with its serum proteins was histopathologically observed in the present work for the first time. The cooperative use of P. flavoviridis antivenom and its serum proteins as medication for P. flavoviridis snake bites is discussed. PMID:24139850

Chijiwa, Takahito; So, Shuhei; Hattori, Shosaku; Yoshida, Aichi; Oda-Ueda, Naoko; Ohno, Motonori

2013-12-15

133

Determination of serum protein binding affinity of inhibitors from analysis of concentration-response plots in biochemical activity assays.  

PubMed

Serum protein binding is a common problem with synthetic molecules designed as enzyme and receptor inhibitors for in vivo clinical use. The theoretical basis of a simple method is described. In this method, the dissociation constant for serum protein binding may be determined from analysis of the shift in apparent IC(50) (concentration at which 50% inhibition of activity is observed) caused by the presence of varying concentrations of serum (or individual serum proteins) in biochemical activity assays. Knowledge of the serum protein dissociation constant and the serum concentration of the binding protein can be used to predict the amount of free compound available in vivo after dosing to achieve a specific total concentration of compound in the blood stream. PMID:10906723

Copeland, R A

2000-08-01

134

Supplementation of orotic acid to the casein, but not to egg protein, soy protein, or wheat gluten diets, increases serum ornithine carbamoyltransferase activity  

Microsoft Academic Search

Effects of dietary supplementation of orotic acid to a diet containing the casein protein were compared with diets containing egg protein, soy protein, or wheat gluten on lipid levels in the liver and serum and activities of ornithine carbamoyltransferase (OCT) and alanine aminotransferase in the serum of rats. We found that supplementation of orotic acid to each diet increased the

Yoritaka Aoyama; Mizuho Wada

2000-01-01

135

Protein intake, control of serum phosphorus, and relatively low levels of parathyroid hormone in elderly hemodialysis patients.  

PubMed

In maintenance hemodialysis (MHD) patients, advanced age is a factor associated with relative hypoparathyroidism and an adynamic bone. However, the underlying mechanism remains elusive. The aim of the present study was to analyze whether the observed spontaneous decrease in protein intake in the elderly favors a better control of serum phosphorus (P) and parathyroid hormone (PTH) levels. A cross-sectional study including 207 MHD patients (mean age, 60 +/- 14; 59% males; time on dialysis, 61 +/- 55 months) dialyzed 3.5 to 4.5 hours 3 times a week using bicarbonate hemodialysis from 6 Spanish hemodialysis centers was performed. In 95 patients, the nutrient intake was recorded over a 5-day period, and average daily ingestion of nutrients was calculated using a computerized diet analysis system. One-way analysis of variance showed that serum phosphorus and intact PTH decreased with age. In addition, patients with serum phosphorus lower than 4 mg/dL as compared with those with serum phosphorus greater than 4 mg/dL were older (68 +/- 9 v 58 +/- 15 years, P < 0.001), had a lower protein (0.86 +/- 0.3 v 1.05 +/- 0.4 g/kg body weight, P < 0.01) and caloric intake (21.9 +/- 7.4 v 25.7 +/- 8.3 kcal/Kg body weight, P < 0.01), and had lower PTH levels (102 +/- 155 v 290 +/- 345 pg/mL, P < 0.001). An inverse and significant correlation was observed between age and protein intake (r = -0.48; P < 0.01), caloric intake (r = -0.37, P < 0.01), serum phosphorus concentration (r = -0.40; P < 0.01), and PTH levels (r = -0.26; P < 0.01). Additionally, a significant positive correlation was found between serum phosphorus and PTH levels (r = 0.40; P < 0.01). The results obtained in the present study suggest that a lower serum phosphorus level due to spontaneous reduction of protein intake might contribute to the relative low PTH levels observed in elderly hemodialysis patients. PMID:11382697

Lorenzo, V; Martín, M; Rufino, M; Jiménez, A; Malo, A M; Sanchez, E; Hernández, D; Rodríguez, M; Torres, A

2001-06-01

136

Canine cancer screening via ultraviolet absorbance and fluorescence spectroscopy of serum proteins  

NASA Astrophysics Data System (ADS)

A cost-effective optical cancer screening and monitoring technique was demonstrated in a pilot study of canine serum samples and was patented for commercialization. Compared to conventional blood chemistry analysis methods, more accurate estimations of the concentrations of albumin, globulins, and hemoglobin in serum were obtained by fitting the near UV absorbance and photoluminescence spectra of diluted serum as a linear combination of component reference spectra. Tracking these serum proteins over the course of treatment helped to monitor patient immune response to carcinoma and therapy. For cancer screening, 70% of dogs with clinical presentation of cancer displayed suppressed serum hemoglobin levels (below 20 mg/dL) in combination with atypical serum protein compositions, that is, albumin levels outside of a safe range (from 4 to 8 g/dL) and globulin levels above or below a more normal range (from 1.7 to 3.7 g/dL). Of the dogs that met these criteria, only 20% were given a false positive label by this cancer screening test.

Dickerson, Bryan D.; Geist, Brian L.; Spillman, William B., Jr.; Robertson, John L.

2007-11-01

137

Comparative analysis of serum proteomes: Identification of proteins associated with sciatica due to lumbar intervertebral disc herniation  

PubMed Central

Lumbar intervertebral disc herniation (LDH) is one of the most common orthopedic conditions that can cause lower back pain and sciatica. However, the pathogenesis of LDH is poorly understood. The aim of the present study was to use proteomic analysis of blood samples to establish whether there are serum proteins associated with LDH, which may be useful in elucidating LDH pathogenesis. The ultimate aim was to develop a simple technique for the diagnosis of LDH based on the blood samples of patients with sciatica. The study used comparative analysis of serum proteomes associated with sciatica due to LDH. A total of 30 LDH patients with sciatica, receiving treatment between August and December 2007, were selected as the experimental group (or LDH group). A total of 2 ml of blood was obtained from each of the 30 patients in the LDH group and from 30 healthy volunteers, who constituted the control group. Two-dimensional electrophoresis of the blood samples was conducted, distinct protein spots were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and proteins associated with LDH were detected. An enzyme-linked immunosorbent assay (ELISA) was developed to screen for the LDH proteins and was tested on the sera of a second test and control group that included 10 patients with LDH and 10 healthy subjects, respectively. Based on signal intensity, the expression levels of 6 proteins on the dielectrophoretogram were found to be significantly associated with LDH. The identities of the LDH proteins were upregulated apolipoprotein-L1 (APO-L1) and two types of serum albumin precursors, and downregulated apolipoprotein M (APO-M), tetranectin (TN) and immunoglobulin light chain (IGL). Further ELISA experiments confirmed that there were increased serum levels of 4 out of the 6 proteins in patients with sciatica due to LDH, which was statistically different compared to the healthy subjects. In conclusion, these results suggest that serum APO-L1, TN, APO-M and IGL may serve as LDH biomarkers. PMID:25054013

XIE, PEIGEN; LIU, BIN; CHEN, RUIQIANG; YANG, BU; DONG, JIANWEN; RONG, LIMIN

2014-01-01

138

Evaluation of serum biochemical marker concentrations and survival time in dogs with protein-losing enteropathy.  

PubMed

Objective-To evaluate serum concentrations of biochemical markers and survival time in dogs with protein-losing enteropathy (PLE). Design-Prospective study. Animals-29 dogs with PLE and 18 dogs with food-responsive diarrhea (FRD). Procedures-Data regarding serum concentrations of various biochemical markers at the initial evaluation were available for 18 of the 29 dogs with PLE and compared with findings for dogs with FRD. Correlations between biochemical marker concentrations and survival time (interval between time of initial evaluation and death or euthanasia) for dogs with PLE were evaluated. Results-Serum C-reactive protein concentration was high in 13 of 18 dogs with PLE and in 2 of 18 dogs with FRD. Serum concentration of canine pancreatic lipase immunoreactivity was high in 3 dogs with PLE but within the reference interval in all dogs with FRD. Serum ?1-proteinase inhibitor concentration was less than the lower reference limit in 9 dogs with PLE and 1 dog with FRD. Compared with findings in dogs with FRD, values of those 3 variables in dogs with PLE were significantly different. Serum calprotectin (measured by radioimmunoassay and ELISA) and S100A12 concentrations were high but did not differ significantly between groups. Seventeen of the 29 dogs with PLE were euthanized owing to this disease; median survival time was 67 days (range, 2 to 2,551 days). Conclusions and Clinical Relevance-Serum C-reactive protein, canine pancreatic lipase immunoreactivity, and ?1-proteinase inhibitor concentrations differed significantly between dogs with PLE and FRD. Most initial biomarker concentrations were not predictive of survival time in dogs with PLE. PMID:25517330

Equilino, Mirjam; Théodoloz, Vincent; Gorgas, Daniela; Doherr, Marcus G; Heilmann, Romy M; Suchodolski, Jan S; Steiner, Jörg M; Burgener Dvm, Iwan A

2015-01-01

139

Serum proteins of neonatal pigs orally inoculated with Isospora suis oocysts.  

PubMed

Serum proteins were evaluated by agarose electrophoresis at periodic intervals between postinoculation days (PID) 2 and 35 in 4 litters of pigs inoculated orally at 36 to 38 hours of age with 70,000 sporulated Isospora suis oocysts and in 4 litters of age-matched noninoculated control pigs. In inoculated pigs, clinical disease characterized by vomiting and diarrhea began at PID 3 to 4 and was resolved by PID 11. Isospora suis oocysts were detected in feces of inoculated pigs from PID 5 to 25, with peak numbers present on PID 5 to 6. Of 43 pigs inoculated with I suis oocysts, 4(9.3%) died of coccidiosis, and samples were not obtained for serum protein evaluation. Of 39 noninoculated control pigs, all remained clinically normal, and I suis oocysts were not detected in their feces. Serum protein fraction values in inoculated and control groups compared at each sample collection time did not differ significantly, except at PID 15, when beta-globulin values were lower in inoculated pigs (P = 0.02). At PID 35, total serum proteins, albumin, and alpha 1-, beta-, and gamma-globulin values of inoculated pigs were lower than those of controls, but there were too few pigs examined for definitive statistical analysis. Differences in trends over time were observed between inoculated and control groups for several serum protein fractions. In inoculated pigs, total proteins (P less than 0.1) and beta-globulins (P less than 0.01) decreased with time, whereas those of control pigs increased. Similar differences in trends were noticed for albumin and alpha 1-globulins, but these were not significant.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:3358549

Jarvinen, J A; Zimmerman, G L; Schons, D J; Guenther, C

1988-03-01

140

Prooligonucleotides exhibit less serum-protein binding than phosphodiester and phosphorothioate oligonucleotides.  

PubMed

The protein-binding properties of dodecathymidine derivatives (prooligos) bearing either methyl- or tert-butyl-S-acylthioethyl (Me- or tBuSATE) protecting groups were evaluated. The dissociation constants (Kd) were estimated for complexes of prooligos with serum blood proteins and lactoferrin using prooligos to compete the binding of the radiolabeled, alkylating probe oligonucleotide CIRp(T)12 with the proteins. tBuSATE and MeSATE prooligos have decreased affinity of binding with serum proteins and lactoferrin compared with their parent oligos. These data suggest that prooligos should cause less side effects which combined with their stability to nucleases and enhanced permeability into cells make them potentially useful for design of therapeutics. PMID:10893717

Tosquellas, G; Bryksin, A; Alvarez, K; Morvan, F; Vasseur, J J; Rayner, B; Rykova, E; Laktionov, P; Vlassov, V; Imbach, J L

2000-01-01

141

High-resolution two-dimensional protein electrophoresis of pathological plasma/serum.  

PubMed

The potential usefulness of an optimalized high-resolution two-dimensional gel electrophoresis (2-DGE) protocol was studied by comparative analysis of plasma/serum obtained from apparently healthy individuals and from patients with a few selected known diseases. Despite their apparent complexity, patient electrophoretograms revealed readily detectable modifications of the 'reference' protein profile for those selected diseases (listed below). Abnormal profiles were characterized by presence or absence of particular spots, by reduction or enlargement of spot size, or by alterations of spot microheterogeneity. Combinations of several modifications enabled different 'disease-associated spot pattern' to be distinguished on the protein maps of patients with: monoclonal gammopathies, hypogammaglobulinemia, hepatic failure, chronic renal failure and hemolytic anemia. This study demonstrates that identification of plasma/serum protein alterations by 2-DGE allows a few selected diseases to be diagnosed solely on the basis of protein map modifications. PMID:1932211

Tissot, J D; Schneider, P; James, R W; Daigneault, R; Hochstrasser, D F

1991-01-01

142

Selective precipitation of haptoglobin and alpha2-macroglobulin from human serum using Alocasia macrorhiza tuber protein.  

PubMed

Treatment of human serum with ammonium sulfate fraction (0-50%) of Alocasia macrorhiza tuber extract resulted in precipitation at neutral pH. The precipitate was dissolved at pH 10.5 and chromatographed on Sephadex G-100 column. Two protein peaks were resolved. While the first peak represented alpha2-macroglobulin and haptoglobin, the second peak accounted for specific Alocasia protein. Incidentally the Alocasia protein was shown to be responsible for selective and specific precipitation of alpha2-macroglobulin and haptoglobin from serum. Thus the plant protein in its pure form or in crude stage could be used for the rapid isolation of two of the prominent alpha2-globulins. PMID:12553858

Nayak, B Shivananda; Ulloor, N Jagadish; Shivaraj, B

2002-12-01

143

The development of simple and sensitive small-molecule fluorescent probes for the detection of serum proteins after native polyacrylamide gel electrophoresis.  

PubMed

In this paper, a simple and sensitive small-molecule fluorescent probe, 2,5-dihydroxy-4'-dimethylaminochalcone (DHDMAC), was designed and synthesized for the detection of human serum proteins via hydrophobic interactions after polyacrylamide gel electrophoresis (PAGE). This probe produced lower fluorescence emission in the absence of proteins, and the emission intensity was significantly increased after the interaction with serum proteins. To demonstrate the imaging performance of this probe as a fluorescent dye, a series of experiments was conducted that included sensitivity comparison and 2D-PAGE. The results indicated that the sensitivity of DHDMAC staining is comparable to that of the most widely used fluorescent dye, SYPRO Ruby, and more protein spots (including thyroxine-binding globulin, angiotensinogen, afamin, zinc-?-2-glycoprotein and ?-1-antichymotrypsin) were detected after 2D-PAGE. Therefore, DHDMAC is a good protein reporter due to its fast staining procedure, low detection limits and high resolution. PMID:22475746

Wang, Fangfang; Huang, Lingyun; Na, Na; He, Dacheng; Sun, Dezhi; Ouyang, Jin

2012-05-21

144

Serum Levels of Calcification Inhibition Proteins and Coronary Artery Calcium Score: Comparison between Transplantation and Dialysis  

Microsoft Academic Search

Vascular calcifications in CKD are now linked to serum alterations of both divalent ions and calcification inhibitory proteins. Due to possible biochemical differences between dialysis (D) and transplantation (Tx), we examined the entity and severity of these biochemical modifications and of coronary artery calcium score separately in these two populations. We assayed, besides standard markers of inflammation, divalent ions and

Sandro Mazzaferro; Marzia Pasquali; Francesco Pugliese; Giusi Barresi; Iacopo Carbone; Marco Francone; Daniela Sardella; Franco Taggi

2007-01-01

145

SERUM PROTEINS AND THEIR FRACTIONS IN THE TIMAHDITE SHEEP IN MOROCCO  

E-print Network

with age. There was no significant difference between 2 year-old animals and 3-4 year-old onesSERUM PROTEINS AND THEIR FRACTIONS IN THE TIMAHDITE SHEEP IN MOROCCO : VARIATIONS WITH AGE months-old, 2 years-old, 3-4 years-old and more than 4 years-old. Influence of the season was studied

Paris-Sud XI, Université de

146

CHANGES IN SERUM PROTEINS OF RATS SUBJECTED TO SMALL DOSES OF X RAYS. II. GLYCOPROTEINS  

Microsoft Academic Search

As in earlier studies on total proteins (cf. preceding abstract), rats ; were exposed to whole-body or abdominal irradiation with low doses, 50 to 80 r ; given in single or four fractionated doses over 4 days. At 24 hr and 15 days ; after irradiation serum samples were obtained and treated with 95% ethanol, the ; glycoproteins remaining in

F. A. Paolini; A. Pannella

1961-01-01

147

Comparative study of modifications produced by x rays in bovine serum proteins  

Microsoft Academic Search

Serum albumin and gamma globulin appear to be more radiosensitive than ; alpha and BETA globulins. Small irradiation doses are sufficient to ; chemically modify certain sites in these proteins without producing very serious ; modifications in the secondary and tertiary structures. Such modifications ; contribute to the understanding of the differences in the physiological, and in ; particular antigen

J. L. Azanza; A. Ducastaing; J. Raymond; P. Creach

1972-01-01

148

Changes of Growth and of Serum Proteins in Ducklings Intoxicated with Cobalt  

Microsoft Academic Search

Changes of growth and serum proteins were found in ducklings nourished with commercial diet enriched with cobalt. It was found that a high concentration of this element in diet involves a significant diminution of growth, an increase of ?-globulins and a decrease of albumin. It was observed too that the diminution of concentration of cobalt in diet involves after- wards

S. Paulov

1971-01-01

149

Serum protein changes in immune and nonimmune pigeons infected with various strains of Trichomonas gallinae  

USGS Publications Warehouse

Serum protein changes were studied in immune and nonimmune pigeons infected with three different strains of Trichomonas gallinae. Strain I (nonvirulent) produced no change in the relative concentration of serum components. Strains II (oral canker) and III (Jones' Barn) produced decreases in albumin and alpha globulins, and increases in beta and gamma globulins between the 7th and 20th days post infection. Birds infected with strain II began to return to normal by the 20th day, while all those infected with strain III were dead between 10 and 14 days post infection. Two serum protein patterns resulted from infection of immune birds with the Jones' Barn strain. One showed no change in relative protein concentrations and no tissue invasion by the parasite while the other was similar to that seen in nonimmune birds infected with a strain producing oral canker. These also showed evidence of tissue invasion by the parasite. It was concluded that tissue invasion was necessary to evoke a quantitative change in serum protein concentrations.

Kocan, R.M.; Herman, C.M.

1970-01-01

150

Peculiarities of the Genetic Variability of Blood Serum Proteins in Some Species of the Family Anatidae  

Microsoft Academic Search

We investigated blood serum of the following eight waterfowl species from the family Anatidae in polyacrylamide gel: Goosander (Mergus merganser), Smew (Mergus albellus), Eider (Somateria mollissima), Spectacled Eider (Somateria fisheri), Common Scoter (Melanitta nigra), Harlequin (Histronicus histronicus), Goldeneye (Bucephala clangula) and Long-tailed Duck (Clangula hyemalis). During our investigations we identified the systems of general protein and enzymes in polyacrylamide gel.

Sigita Slav?nait?; Aniolas Sruoga; Algimantas Paulauskas; Elena Mozalien?

2000-01-01

151

The effects of temperature on thyroid hormone binding to serum proteins in sea turtles  

E-print Network

THE EFFECTS OF TEMPERATURE ON THYROID HORMONE BINDING TO SERUN PROTEINS IN SEA TURTLES A Thesis by SHANE PATRICK HAYNES Submitted to the Office of Graduate Studies of Texas A&M University in partial fulfillment of the requirements... for the degree of MASTER OF SCIENCE August 1990 Major Subject: Zoology THE EFFECTS OF TEMPERATURE ON THYROID HORMONE BINDING TO SERUM PROTEINS IN SEA TURTLES A Thesis by SHANE PATRICK HAYNES Approved as to style and content by: Duncan S. Mac...

Haynes, Shane Patrick

1990-01-01

152

Growth hormone receptor and serum binding protein: purification, cloning and expression  

Microsoft Academic Search

A putative growth hormone receptor from rabbit liver and the growth hormone binding protein from rabbit serum have the same ammo-terminal amino-acid sequence, indicating that the binding protein corresponds to the extracellular hormone-binding domain of the liver receptor. The complete amino-acid sequences derived from complementary DNA clones encoding the putative human and rabbit growth hormone receptors are not similar to

David W. Leung; Steven A. Spencer; George Cachianes; R. Glenn Hammonds; Carol Collins; William J. Henzel; Ross Barnard; Michael J. Waters; William I. Wood

1987-01-01

153

Serum protein profiles as potential biomarkers for infectious disease status in pigs  

PubMed Central

Background In veterinary medicine and animal husbandry, there is a need for tools allowing the early warning of diseases. Preferably, tests should be available that warn farmers and veterinarians during the incubation periods of disease and before the onset of clinical signs. The objective of this study was to explore the potential of serum protein profiles as an early biomarker for infectious disease status. Serum samples were obtained from an experimental pig model for porcine circovirus-associated disease (PCVAD), consisting of Porcine Circovirus type 2 (PCV2) infection in combination with either Porcine Parvovirus (PPV) or Porcine Reproductive and Respiratory Syndrome virus (PRRSV). Sera were collected before and after onset of clinical signs at day 0, 5 and 19 post infection. Serum protein profiles were evaluated against sera from non-infected control animals. Results Protein profiles were generated by SELDI-TOF mass spectrometry in combination with the Proteominer™ technology to enrich for low-abundance proteins. Based on these protein profiles, the experimentally infected pigs could be classified according to their infectious disease status. Before the onset of clinical signs 88% of the infected animals could be classified correctly, after the onset of clinical sigs 93%. The sensitivity of the classification appeared to be high. The protein profiles could distinguish between separate infection models, although specificity was moderate to low. Classification of PCV2/PRRSV infected animals was superior compared to PCV2/PPV infected animals. Limiting the number of proteins in the profiles (ranging from 568 to 10) had only minor effects on the classification performance. Conclusions This study shows that serum protein profiles have potential for detection and identification of viral infections in pigs before clinical signs of the disease become visible. PMID:22439879

2012-01-01

154

Novel serum protein biomarker panel revealed by mass spectrometry and its prognostic value in breast cancer  

PubMed Central

Introduction Serum profiling using proteomic techniques has great potential to detect biomarkers that might improve diagnosis and predict outcome for breast cancer patients (BC). This study used surface-enhanced laser desorption/ionization time-of-flight (SELDI-TOF) mass spectrometry (MS) to identify differentially expressed proteins in sera from BC and healthy volunteers (HV), with the goal of developing a new prognostic biomarker panel. Methods Training set serum samples from 99 BC and 51 HV subjects were applied to four adsorptive chip surfaces (anion-exchange, cation-exchange, hydrophobic, and metal affinity) and analyzed by time-of-flight MS. For validation, 100 independent BC serum samples and 70 HV samples were analyzed similarly. Cluster analysis of protein spectra was performed to identify protein patterns related to BC and HV groups. Univariate and multivariate statistical analyses were used to develop a protein panel to distinguish breast cancer sera from healthy sera, and its prognostic potential was evaluated. Results From 51 protein peaks that were significantly up- or downregulated in BC patients by univariate analysis, binary logistic regression yielded five protein peaks that together classified BC and HV with a receiver operating characteristic (ROC) area-under-the-curve value of 0.961. Validation on an independent patient cohort confirmed the five-protein parameter (ROC value 0.939). The five-protein parameter showed positive association with large tumor size (P?=?0.018) and lymph node involvement (P?=?0.016). By matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) MS, immunoprecipitation and western blotting the proteins were identified as a fragment of apolipoprotein H (ApoH), ApoCI, complement C3a, transthyretin, and ApoAI. Kaplan-Meier analysis on 181 subjects after median follow-up of >5 years demonstrated that the panel significantly predicted disease-free survival (P?=?0.005), its efficacy apparently greater in women with estrogen receptor (ER)-negative tumors (n?=?50, P?=?0.003) compared to ER-positive (n?=?131, P?=?0.161), although the influence of ER status needs to be confirmed after longer follow-up. Conclusions Protein mass profiling by MS has revealed five serum proteins which, in combination, can distinguish between serum from women with breast cancer and healthy control subjects with high sensitivity and specificity. The five-protein panel significantly predicts recurrence-free survival in women with ER-negative tumors and may have value in the management of these patients. PMID:24935269

2014-01-01

155

THE SERUM PROTEIN LEVELS IN RATS IRRADIATED WITH Co⁶° AND TREATED WITH HOMOLOGOUS HOMOGENATES OF SPLEEN AND LIVER  

Microsoft Academic Search

Serum proteins were studied by paper electrophoresis in rats after ; irradiation with 450 r, either in 2 daily doses of 225 r or in a single dose. ; Eight days after the single dose, total serum protein levels were unaffected but ; albumin fell to 1.82 g% from a control level of 2.64, alpha â-globulin ; increased to 1.06

P. Erede; A. M. Podesta

1961-01-01

156

THE ELECTROPHORETIC PATTERN OF SERUM PROTEINS IN NEOPLASTIC DISEASES, WITH PARTICULAR REGARD TO PATIENTS SUBJECTED TO RADIOTHERAPY  

Microsoft Academic Search

Serum protein fractions were quantitatively determined in 104 patients ; with lymphoblastomas or carcinomas before and after radiotherapy. Irradiation ; tended to normalize the serum protein levels, which before irradiation consisted ; in a reduction in albumin and elevations in alpha â- and gamma -globulins. ; (BBB);

Magno

1961-01-01

157

Ratio of serum tartrate-inhibitable acid phosphatase to total serum protein in benign prostatic hypertrophy and prostatic carcinoma.  

PubMed

The activity concentration and the specific activity (the ratio of enzyme activity to total serum protein) of the tartrate-inhibitable fraction of acid phosphatase [orthophosphoric monoester phosphohydrolase (acid optimum), EC 3.1.3.2; TIAP] were related to benign prostatic hypertrophy and to prostatic carcinoma. As expected, the TIAP activity concentrations assayed in the sera of patients with benign prostatic hypertrophy were within the range of those assayed in normal human sera. In contrast, the specific activities of TIAP determined in the sera of patients with benign prostatic hypertrophy were significantly higher than those determined in the control group. In the sera of prostatic carcinoma patients, both the TIAP activity concentrations and the TIAP specific activities differed significantly (F = 730) from the normal values. PMID:1371723

Petrovich, G; Ursich-Jankovich, J; Prijovich, Z

1992-02-01

158

Serum C-reactive protein level but not its gene polymorphism is associated with Takayasu arteritis.  

PubMed

Takayasu arteritis (TA) patients with active disease often have elevated serum C-reactive protein (CRP) levels, which usually decline with the disease remission. The serum CRP concentration has been showed to be related to CRP gene polymorphisms in previous studies. The present study aims to investigate the associations of serum level of CRP and CRP polymorphisms with TA. A total of 178 unrelated Chinese Han TA patients and 229 unrelated Chinese Han individuals without documented disease were enrolled in our studies. After a systemic search in the HapMap database, four single-nucleotide polymorphisms (SNPs) were selected, namely, rs1800947, rs3093077, rs1205, and rs2808630. The ligase detection reaction (LDR) was used in genotyping. CRP concentrations were determined using turbidimetric immunoassay. Genotype frequencies and allele frequencies of CRP variations were similar between TA patients and controls. CRP haplotype frequencies in patients were not significantly different from those of controls. No significant association between serum CRP concentrations and genotypes was found. Moreover, no association was found in CRP concentration between patients with types I, II, and III TA or between patients with or without pulmonary involvement. By contrast, serum CRP concentration was directly correlated with disease severity. In conclusion, CRP polymorphisms were not associated with TA susceptibility or serum CRP levels in the Chinese Han population. However, higher CRP level was correlated with a more serious disease status, which implies that CRP possibly contributes to the progression of TA. PMID:24894103

Cheng, Yanmei; Dang, Aimin; Lv, Naqiang; Gao, Qian; Chen, Bingwei; Liu, Guozhang

2014-06-01

159

The stress of weaning influences serum levels of acute-phase proteins, iron-binding proteins, inflammatory cytokines, cortisol, and leukocyte subsets in Holstein calves  

PubMed Central

The purpose of our study was to investigate changes in immunological parameters induced by weaning stress (including milk restriction) in calves. Fifteen Holstein calves were subjected to weaning at 6 weeks of age. Blood samples were collected at -14, -7, -2, 1, 3, and 5 days post-weaning (DPW; 0 DPW = 42 days). Weaning caused significant (p < 0.01) increases in the neutrophil (NE):lymphocyte (LY) ratio at 5 DPW with a significant (p < 0.05) reduction of LYs. The concentration of acute-phase proteins (haptoglobin and serum amyloid A) also increased significantly (p < 0.05) at 3 and 5 DPW compared to -2 DPW. Levels of the iron-binding protein lactoferrin decreased significantly (p < 0.05) after weaning. Serum tumor necrosis factor-? and cortisol levels were elevated (p < 0.05) at 3 DPW, while those of serum interferon-? decreased (p < 0.05) at 1 and 3 DPW compared to levels observed before weaning. Weaning significantly (p < 0.05) decreased the percentage of CD25+ T cells in the peripheral blood. In conclusion, weaning stress affected the NE:LY ratio along with the levels of acute phase proteins, lactoferrin, cortisol, and inflammatory cytokines in the peripheral blood of calves. Weaning stress may induce an acute phase response possibly through the elevation of cortisol production and modulation of inflammatory cytokines. PMID:21586874

Kim, Myung-Hoo; Yang, Ji-Young; Upadhaya, Santi Devi; Lee, Hyun-Jun; Yun, Cheol-Heui

2011-01-01

160

Mid-Trimester Maternal Serum hCG and Alpha Fetal Protein Levels: Clinical Significance and Prediction of Adverse Pregnancy Outcome  

PubMed Central

Context Maternal serum human Chorionic Gonadotropin (hCG) and Alpha Fetal Protein (AFP) were originally introduced to detect trisomy 21 and neural tube defects. However, in the absence of aneuploidy or neural tube defects, mid-trimester maternal serum hCG and/or maternal serum AFP associated with adverse pregnancy outcomes. Pregnancies with unexplained mid-trimester elevation in maternal serum hCG and/or maternal serum AFP, are at increased risk for pregnancy complications resulting from placental insufficiency. Evidence Acquisition Mid-trimester maternal serum hCG>2.5 MoM associated with an increased risk for pregnancy complications including: late fetal loss, gestational hypertension, preeclampsia, intrauterine growth restriction (IUGR), preterm delivery and intrauterine fetal death(IUFD). Mid-trimester maternal serum AFP levels >2.5 MoM are thought to reflect a defect in placentation and associated with an increased risk for pregnancy complications including: late fetal loss, gestational hypertension, preeclampsia, IUGR, preterm delivery and IUFD. Results Combined mid-trimester elevation in maternal serum hCG and AFP levels suggest a more complex type of placental pathology. They have stronger association with pregnancy complications including: late fetal loss, gestational hypertension, preeclampsia, IUGR, preterm delivery and IUFD. Conclusions Mid-trimester maternal serum hCG or AFP levels alone cannot detect all pregnant women with increased risk to develop pregnancy complications. Multiparameter testing of placental function in mid-trimester (maternal serum hCG and AFP screening, uterine artery Doppler and placental morphology) may allow us to identify women with increased risk to develop severe placental insufficiency and pregnancy complications. However, future prospective studies are needed to confirm the prognostic significance of multiparameter testing of placental function in mid-trimester. PMID:23825981

Androutsopoulos, Georgios; Gkogkos, Panagiotis; Decavalas, Georgios

2013-01-01

161

Ferritin L is the sole serum ferritin constituent and a positive hepatic acute-phase protein.  

PubMed

Ferritin L (FTL) and ferritin H (FTH) subunits are responsible for intracellular iron storage. Serum ferritin levels are not only dependant on body iron stores. Aims of the present study are to demonstrate nature, source, and major regulatory mediators of serum ferritin in an animal model of acute-phase (AP) response. Animals (rats, wild-type [WT] mice, and interleukin [IL]-6ko mice) were injected with turpentine oil (TO) intra-muscularity to induce a sterile abscess and sacrificed at different time points afterward. Rat hepatocytes were isolated for cell culture and, after reaching confluence, stimulated with major AP cytokines to induce AP conditions. We found a significantly increased expression of both ferritin subunits in liver at mRNA and protein levels during AP response. In the serum of both control and TO-injected rats, only FTL was detectable by Western blotting, whereas no increase in serum FTL was measured by Western blot or enzyme-linked immunosorbent assay. An increase in protein expression of FTL and FTH was observed in lysates of rat hepatocytes after treatment with IL-6, IL-1?, and tumor necrosis factor-?; however, only FTL was increasingly released into supernatant. In both TO-injected rats and WT mice, a dramatic increase in serum IL-6 levels was observed, along with an increased amount of hepatic ferritin subunits. However, an increase of hepatic FTL but not of FTH protein expression was observed in IL-6ko mice after TO injection. Our data demonstrate that FTL is the only rat serum ferritin whose release into circulation from the hepatocytes is increased by the effect of AP cytokines (e.g., IL-6). In contrast, FTH expression is intracellular in both under physiological and AP conditions. PMID:23524846

Naz, Naila; Moriconi, Federico; Ahmad, Shakil; Amanzada, Ahmad; Khan, Sajjad; Mihm, Sabine; Ramadori, Guiliano; Malik, Ihtzaz Ahmed

2013-06-01

162

Binding of labeled thyroxin analog to serum proteins evaluated after radioimmunoassay of free thyroxin  

SciTech Connect

In ambulatory patients, assay of free thyroxin (FT4) in serum correlates well with thyroid status and with results obtained by equilibrium dialysis. The validity of FT4 results has been questioned mainly in euthyroid patients with altered concentrations of thyroid hormone-binding proteins, as in nonthyroidal illness, hereditary analbuminemia, familial dysalbuminemic hyperthyroxinemia (FDH), and the presence of iodothyronine-binding antibodies. I present here a study of the binding of (/sup 125/I)T4-derivative to serum proteins in the supernate, which is ordinarily discarded after determination of FT4 by one-step radioimmunoassay with dextran-coated charcoal used to separate the free and bound fractions. The results are expressed as a ratio, with results for a normal serum pool as reference. The average ratio was high in hyperthyroid subjects, 1.26 (SD 0.12, n = 25), and in hypoalbuminemia, 1.20 (SD 0.10, n = 15), and low in FDH, 0.62 (SD 0.11, n = 9), and hypothyroid subjects, 0.90 (SD 0.06, n = 20). In normal individuals it was 0.98 (SD 0.05, n = 30). Determination of the analog-binding rate complements the FT4 result and allows for the recognition of cases with abnormal binding by serum proteins, without recourse to other tests recommended for thyroid-function studies.

Arevalo, G.

1989-03-01

163

Serum-dependent expression of promyelocytic leukemia protein suppresses propagation of influenza virus  

SciTech Connect

The rate of propagation of influenza virus in human adenocarcinoma Caco-2 cells was found to negatively correlate with the concentration of fetal bovine serum (FBS) in the culture medium. Virus replicated more rapidly at lower FBS concentrations (0 or 2%) than at higher concentrations (10 or 20%) during an early stage of infection. Basal and interferon (IFN)-induced levels of typical IFN-inducible anti-viral proteins, such as 2',5'-oligoadenylate synthetase, dsRNA-activated protein kinase and MxA, were unaffected by variation in FBS concentrations. But promyelocytic leukemia protein (PML) was expressed in a serum-dependent manner. In particular, the 65 to 70 kDa isoform of PML was markedly upregulated following the addition of serum. In contrast, other isoforms were induced by IFN treatment, and weakly induced by FBS concentrations. Immunofluorescence microscopy indicated that PML was mainly formed nuclear bodies in Caco-2 cells at various FBS concentrations, and the levels of the PML-nuclear bodies were upregulated by FBS. Overexpression of PML isoform consisting of 560 or 633 amino acid residues by transfection of expression plasmid results in significantly delayed viral replication rate in Caco-2 cells. On the other hand, downregulation of PML expression by RNAi enhanced viral replication. These results indicate that PML isoforms which are expressed in a serum-dependent manner suppress the propagation of influenza virus at an early stage of infection.

Iki, Shigeo [Department of Microbiology, Sapporo Medical University School of Medicine, Chuo-ku, Sapporo 060-8556 (Japan); Hokkaido Institute of Public Health, Kita-ku, Sapporo 060-0819 (Japan); Yokota, Shin-ichi [Department of Microbiology, Sapporo Medical University School of Medicine, Chuo-ku, Sapporo 060-8556 (Japan); Okabayashi, Tamaki [Department of Microbiology, Sapporo Medical University School of Medicine, Chuo-ku, Sapporo 060-8556 (Japan); Yokosawa, Noriko [Department of Microbiology, Sapporo Medical University School of Medicine, Chuo-ku, Sapporo 060-8556 (Japan); Nagata, Kyosuke [Department of Infection Biology, Graduate School of Comprehensive Human Sciences and Institute of Basic Medical Sciences, University of Tsukuba, Tsukuba 305-8575 (Japan); Fujii, Nobuhiro [Department of Microbiology, Sapporo Medical University School of Medicine, Chuo-ku, Sapporo 060-8556 (Japan)]. E-mail: fujii@sapmed.ac.jp

2005-12-05

164

Immunoglobulin production in the European pond tortoise, Emys orbicularis, immunized with serum protein antigens  

PubMed Central

The immunological responses of the European pond tortoise, Emys orbicularis, to BGG and sheep serum proteins indicate that in this tortoise there is no serum component corresponding with mammalian albumin and that both ?G (7S) and ?M (19S) immunoglobulins are involved when antibodies are produced. A prolonged period of ?M antibody production occurs after primary immunization. The separation of the ?M and ?G immunoglobulins was performed using starch block electrophoresis and Sephadex G-200 gel filtration. The separated immunoglobulins were characterized by immunodiffusion and by starch gel, agar gel and immunoelectrophoresis. ImagesFIG. 1-3FIG. 6FIG. 8FIG. 9FIG. 10 PMID:4173673

Lykakis, J. J.

1968-01-01

165

Objective monitoring of disease activity in polyarteritis by measurement of serum C reactive protein concentration.  

PubMed Central

Serial measurements of the serum concentration of C reactive protein were made in 27 patients with polyarteritis over six years. The concentration was invariably raised when the disease was active, even in patients receiving immunosuppressive treatment, and fell rapidly in association with clinical remission induced by immunosuppression. During periods of complete remission, in the absence of any intercurrent condition, the value remained within the normal range. The correlation between C reactive protein concentration and disease activity was much closer than that between erythrocyte sedimentation rate and disease activity. These results indicate that serial measurement of the serum C reactive protein concentration fills the urgent need for an objective index of the activity of polyarteritis and its response to treatment. PMID:6142751

Hind, C R; Savage, C O; Winearls, C G; Pepys, M B

1984-01-01

166

Cellular Binding of Anionic Nanoparticles is Inhibited by Serum Proteins Independent of Nanoparticle Composition.  

PubMed

Nanoparticles used in biological applications encounter a complex mixture of extracellular proteins. Adsorption of these proteins on the nanoparticle surface results in the formation of a "protein corona," which can dominate the interaction of the nanoparticle with the cellular environment. The goal of this research was to determine how nanoparticle composition and surface modification affect the cellular binding of protein-nanoparticle complexes. We examined the cellular binding of a collection of commonly used anionic nanoparticles: quantum dots, colloidal gold nanoparticles, and low-density lipoprotein particles, in the presence and absence of extracellular proteins. These experiments have the advantage of comparing different nanoparticles under identical conditions. Using a combination of fluorescence and dark field microscopy, flow cytometry, and spectroscopy, we find that cellular binding of these anionic nanoparticles is inhibited by serum proteins independent of nanoparticle composition or surface modification. We expect these results will aid in the design of nanoparticles for in vivo applications. PMID:23956836

Fleischer, Candace C; Kumar, Umesh; Payne, Christine K

2013-09-01

167

Protein and microRNA biomarkers from lavage, urine, and serum in military personnel evaluated for dyspnea  

PubMed Central

Background We have identified candidate protein and microRNA (miRNA) biomarkers for dyspnea by studying serum, lavage fluid, and urine from military personnel who reported serious respiratory symptoms after they were deployed to Iraq or Afghanistan. Methods Forty-seven soldiers with the complaint of dyspnea who enrolled in the STudy of Active Duty Military Personnel for Environmental Dust Exposure (STAMPEDE) underwent comprehensive pulmonary evaluations at the San Antonio Military Medical Center. The evaluation included fiber-optic bronchoscopy with bronchoalveolar lavage. The clinical findings from the STAMPEDE subjects pointed to seven general underlying diagnoses or findings including airway hyperreactivity, asthma, low diffusivity of carbon monoxide, and abnormal cell counts. The largest category was undiagnosed. As an exploratory study, not a classification study, we profiled proteins or miRNAs in lavage fluid, serum, or urine in this group to look for any underlying molecular patterns that might lead to biomarkers. Proteins in lavage fluid and urine were identified by accurate mass tag (database-driven) proteomics methods while miRNAs were profiled by a hybridization assay applied to serum, urine, and lavage fluid. Results Over seventy differentially expressed proteins were reliably identified both from lavage and from urine in forty-eight dyspnea subjects compared to fifteen controls with no known lung disorder. Six of these proteins were detected both in urine and lavage. One group of subjects was distinguished from controls by expressing a characteristic group of proteins. A related group of dyspnea subjects expressed a unique group of miRNAs that included one miRNA that was differentially overexpressed in all three fluids studied. The levels of several miRNAs also showed modest but direct associations with several standard clinical measures of lung health such as forced vital capacity or gas exchange efficiency. Conclusions Candidate proteins and miRNAs associated with the general diagnosis of dyspnea have been identified in subjects with differing medical diagnoses. Since these markers can be measured in readily obtained clinical samples, further studies are possible that test the value of these findings in more formal classification or case–control studies in much larger cohorts of subjects with specific lung diseases such as asthma, emphysema, or some other well-defined lung disease. PMID:25282157

2014-01-01

168

Evaluation of steric exclusion chromatography on cryogel column for the separation of serum proteins.  

PubMed

Steric exclusion chromatography (SXC) is a new mode of protein chromatography, in which large proteins are retained on hydrophilic stationary phase surface due to the steric exclusion of polyethylene glycol (PEG) in the mobile phase, and thereafter the retained proteins can be eluted by reducing PEG concentration. In this work, SXC was evaluated on a polyacrylamide cryogel monolith. Microscopic observation of ?-globulin precipitates on the gel surface in SXC was reported for the first time. Due to the compact packing of protein precipitates on the stationary phase surface, the dynamic retention capacity of the cryogel monolith for ?-globulin reached 20 mg/mL bed volume, much higher than those of cryogel beds in adsorption-based chromatography. The effect of molecular weight and concentration of PEG, solution pH and salt concentration on protein retention capacity was in agreement with the earlier work on SXC. Because the cryogel monoliths with interconnected macropores (10-100 ?m) allow much easy flow-through of viscous PEG buffer, the SXC can be operated at low back pressure. Hence, the cryogel monoliths are more suitable for SXC than other monoliths of narrow pores reported previously. In the separation of bovine serum proteins, albumin was recovered in the breakthrough fraction with high purity, and globulin was over eight times concentrated in the elution pool. This work has, thus, demonstrated the rapid serum protein separation and concentration by SXC on the cryogel monolith columns. PMID:24552971

Wang, Chuan; Bai, Shu; Tao, Shi-Peng; Sun, Yan

2014-03-14

169

Association between brain metastasis from lung cancer and the serum level of myelin basic protein  

PubMed Central

The aim of the present study was to determine the association between the expression of myelin basic protein in the serum and the metastasis of lung cancer to the brain. A total of 68 lung cancer patients, treated in the Department of Respiratory Medicine of the People’s Hospital of Rizhao (Rizhao, China), were divided into two groups, those with brain metastasis (32 cases) and those without brain metastasis (36 cases). The expression levels of myelin basic protein were measured for all the patients. The results indicated that the expression levels of myelin basic protein in the brain metastasis group were significantly higher when compared with those in the group without metastasis (P<0.05). However, there was no statistically significant correlation between the size of the brain metastasis and the expression levels of myelin basic protein (P>0.05). Furthermore, no statistically significant difference was found in the average level of myelin basic protein between the two subgroups of patients with brain tumor diameters of >1.5 cm and <1.5 cm (P>0.05). Therefore, the results demonstrated a statistically significant correlation between the expression of myelin basic protein in the serum and the metastasis of lung cancer to the brain. Myelin basic protein may thus prove useful in the early diagnosis of brain metastases in lung cancer patients.

LIU, WEI; ZHAO, JING; WEI, YUJUAN

2015-01-01

170

Expression of serum survivin protein in diagnosis and prognosis of gallbladder cancer: a comparative study.  

PubMed

The role of survivin in gallbladder cancer (GBC) has not been evaluated. We investigated survivin protein expression in serum of patients with gallbladder diseases (cholelithiasis, n = 30; GBC, n = 39) and compared with healthy controls (n = 25). Clinicopathological parameters, diagnosis and prognosis of patients with GBC were correlated with the expression of serum survivin by enzyme-linked immunosorbent assay. Significantly higher (P < 0.0001) expression of survivin protein was observed in GBC as compared to cholelithiasis and control. Increased survivin expression was significantly associated with higher tumor stage (stage III vs. stage II; P < 0.0001) and cellular differentiation (poor and moderate vs. well differentiated; P < 0.0001) in GBC. No significant correlation was observed with any of the other clinico-pathological parameters studied. The cutoff value of survivin protein of 79 pg/ml with sensitivity of 81.16 % and specificity of 80 % differentiated the diseased group (cholelithiasis or GBC) from control group were as the cutoff value of 109 pg/ml differentiated GBC from cholelithiasis with a sensitivity of 82.05 % and specificity of 93.33 %. Though not significant, increased expression of survivin was associated with median overall survival (12 vs. 18 months; P = 0.05) in GBC patients. Our study suggests that survivin protein in serum could be both a useful diagnostic marker and an important prognostic factor for GBC. PMID:25129311

Nigam, Jaya; Chandra, Abhijit; Kazmi, Hasan Raza; Singh, Anshuman; Gupta, Vishal; Parmar, Devendra; Srivastava, Manoj Kumar

2014-09-01

171

Thermodynamic characterization of the interaction between a peptide-drug complex and serum proteins.  

PubMed

The interaction between a peptide-based drug delivery system and two serum proteins, bovine serum albumin (BSA) and immunoglobulin G (IgG), is investigated using fluorescence quenching and calorimetric techniques. An ionic-complementary self/co-assembling peptide, EAR8-II, is employed to encapsulate the hydrophobic anticancer drug pirarubicin (THP) and stabilize it in protein environments. Self/co-assembling properties of the peptide-drug complex (EAR8-II-THP) are shown to be different while interacting with serum proteins compared with the properties of the isolated complex. The results from thermodynamic studies suggest that the drug delivery system has a strong binding affinity (K(SV) 1689 M(-1)), exothermic and enthalpy-driven interaction, with BSA and a relatively weak affinity with IgG (K(SV) 295.2 M(-1)). In the presence of salt ions, the enthalpy and binding affinity remain unchanged, implying other interactions such as hydrogen bonding and Van der Waals interactions are present that are not affected by reduced polarity. This work forms the basis for further studies of EAR8-II-THP complexes in the presence of important proteins and for further evaluation of the complexes' immune response and anticancer activity. PMID:25166955

Sadatmousavi, Parisa; Kovalenko, Eugene; Chen, P

2014-09-23

172

Regulation and Physiological Roles of Serum- and Glucocorticoid-Induced Protein Kinase Isoforms  

NSDL National Science Digital Library

The covalent attachment of phosphate to proteins (phosphorylation), catalyzed by enzymes known as protein kinases, and the removal of phosphate from proteins (dephosphorylation), catalyzed by protein phosphatases, regulate most aspects of cell life. Phosphorylation or dephosphorylation alters the conformation of a protein and can change its ability to function in almost every conceivable way. For example, it cannot only switch activity on or off, but alter the rate at which a protein is degraded or its ability to move from one subcellular compartment to another. There are about 500 protein kinases and 150 protein phosphatases encoded by the human genome, and discovering their biological roles is one of the major tasks of the postgenomic era. This article reviews our current knowledge about one of the protein kinase subfamilies, termed serum- and glucocorticoid-induced protein kinases because the first member (SGK1) was identified as a gene that is rapidly transcribed into mRNA when cells are stimulated by serum or glucocorticoid hormones. However, we now know that the SGK1 gene is transcribed in response to a great variety of extracellular signals. Moreover, the SGK1 enzyme is itself activated by phosphorylation in response to different extracellular signals that act via the formation of a lipid mediator called phosphatidylinositol-3,4,5-trisphosphate (PIP3). Evidence is accumulating that SGK1 plays important roles in activating certain potassium, sodium, and chloride channels, suggesting an involvement in the regulation of processes such as cell survival, the functioning of the brain, and the excretion of sodium by the kidney. For the last mentioned reason, sustained high levels of SGK1 protein and activity may contribute to diseases and conditions, such as hypertension and long-term damage to the kidney in type II diabetes. This raises the possibility that drugs that inhibit SGK1 specifically may have therapeutic potential for the treatment of several diseases.

Florian Lang (University of Tubingen;Department of Physiology REV); Philip Cohen (Scotland;School of Life Sciences, University of Dundee, Dundee REV)

2001-11-13

173

Analysis of s100 calcium binding protein B serum levels in different types of traumatic intracranial lesions.  

PubMed

Abstract The objective of this study was to determine whether the type of intracranial traumatic lesions, the number of simultaneous traumatic lesions, and the occurrence of skull and facial bone fractures have an influence on S100 calcium binding protein B (S100B) serum levels. Patients with blunt traumatic brain injury were prospectively enrolled into this cohort study over a period of 13 months. Venous blood samples were obtained prior to emergency cranial CT scan in all patients within 3?h after injury. The patients were then assigned into six groups: 1) concussion, 2) epidural hematoma, 3) subdural hematoma, 4) subarachnoid hemorrhage, 5) brain contusions, and 6) brain edema. The study included 1696 head trauma patients with a mean age of 57.7±25.3 years, and 126 patients (8%) had 182 traumatic lesions on CT. Significant differences in S100B serum levels were found between cerebral edema and the other four bleeding groups: epidural p=0.0002, subdural p<0.0001, subarachnoid p=0.0001, brain contusions p=0.0003, and concussion p<0.0001. Significant differences in S100B values between patients with one or two intracranial lesions (p=0.014) or with three (p<0.0001) simultaneous intracranial lesions were found. In patients with intracranial traumatic lesions, skull fractures, as well as skull and facial bone fractures occurring together, were identified as significant additional factors for the increase in serum S100B levels (p<0.0001). Older age was also associated with elevated S100B serum levels (p<0.0001). Our data show that peak S100B serum levels were found in patients with cerebral edema and brain contusions. PMID:25068442

Wolf, Harald; Frantal, Sophie; Pajenda, Gholam; Leitgeb, Johannes; Sarahrudi, Kambiz; Hajdu, Stefan

2015-01-01

174

Liver Retinol Transporter and Receptor for Serum Retinol-binding Protein (RBP4)*  

PubMed Central

Vitamin A (retinol) is absorbed in the small intestine, stored in liver, and secreted into circulation bound to serum retinol-binding protein (RBP4). Circulating retinol may be taken up by extrahepatic tissues or recycled back to liver multiple times before it is finally metabolized or degraded. Liver exhibits high affinity binding sites for RBP4, but specific receptors have not been identified. The only known high affinity receptor for RBP4, Stra6, is not expressed in the liver. Here we report discovery of RBP4 receptor-2 (RBPR2), a novel retinol transporter expressed primarily in liver and intestine and induced in adipose tissue of obese mice. RBPR2 is structurally related to Stra6 and highly conserved in vertebrates, including humans. Expression of RBPR2 in cultured cells confers high affinity RBP4 binding and retinol transport, and RBPR2 knockdown reduces RBP4 binding/retinol transport. RBPR2 expression is suppressed by retinol and retinoic acid and correlates inversely with liver retinol stores in vivo. We conclude that RBPR2 is a novel retinol transporter that potentially regulates retinol homeostasis in liver and other tissues. In addition, expression of RBPR2 in liver and fat suggests a possible role in mediating established metabolic actions of RBP4 in those tissues. PMID:23105095

Alapatt, Philomena; Guo, Fangjian; Komanetsky, Susan M.; Wang, Shuping; Cai, Jinjin; Sargsyan, Ashot; Rodríguez Díaz, Eduardo; Bacon, Brandon T.; Aryal, Pratik; Graham, Timothy E.

2013-01-01

175

Spherical Nucleic Acid Nanoparticle Conjugates Enhance G-Quadruplex Formation and Increase Serum Protein Interactions**  

PubMed Central

To understand the effect of three-dimensional oligonucleotide structure on protein corona formation, we studied the identity and quantity of human serum proteins that bind to spherical nucleic acid (SNA) nanoparticle conjugates. SNAs exhibit cellular uptake properties that are remarkably different from those of linear nucleic acids, which have been related to their interaction with certain classes of proteins. Through a proteomic analysis, this work shows that the protein binding properties of SNAs are sequence-specific and supports the conclusion that the oligonucleotide tertiary structure can significantly alter the chemical composition of the SNA protein corona. This knowledge will impact our understanding of how nucleic acid-based nanostructures, and SNAs in particular, function in complex biological milieu. PMID:25393322

Chinen, Alyssa B.; Guan, Chenxia M.

2014-01-01

176

Spherical nucleic Acid nanoparticle conjugates enhance g-quadruplex formation and increase serum protein interactions.  

PubMed

To understand the effect of three-dimensional oligonucleotide structure on protein corona formation, we studied the identity and quantity of human serum proteins that bind to spherical nucleic acid (SNA) nanoparticle conjugates. SNAs exhibit cellular uptake properties that are remarkably different from those of linear nucleic acids, which have been related to their interaction with certain classes of proteins. Through a proteomic analysis, this work shows that the protein binding properties of SNAs are sequence-specific and supports the conclusion that the oligonucleotide tertiary structure can significantly alter the chemical composition of the SNA protein corona. This knowledge will impact our understanding of how nucleic acid-based nanostructures, and SNAs in particular, function in complex biological milieu. PMID:25393322

Chinen, Alyssa B; Guan, Chenxia M; Mirkin, Chad A

2015-01-01

177

Effects of Egg White Protein Supplementation on Muscle Strength and Serum Free Amino Acid Concentrations  

PubMed Central

The aim of this study was to evaluate the effects of egg white protein compared to carbohydrate intake prior to exercise on fat free mass (FFM), one repetition maximum (1RM) muscle strength and blood biochemistry in female athletes. Thirty healthy female collegiate athletes were recruited for this study and matched by sport type, body fat percentage and 1RM leg curl muscle strength. Participants were randomly divided into two groups: protein group (15.0 g egg white protein; 75 kcal) and carbohydrate group (17.5 g maltodextrin, 78 kcal). Supplements were administered daily at the same time in a double-blind manner prior to training during an 8-week period. Measurements were performed before and after the 8-week regimen. The mean dietary energy intake did not change throughout the study period. FFM and 1RM assessments (i.e., leg curl, leg extension, squat, and bench press) increased in both groups. Furthermore, serum urea and serum citrulline levels after the 8-week regimen increased significantly only in the protein group. Our findings indicated that compared to the carbohydrate supplement, the protein supplement was associated with some changes in protein metabolites but not with changes in body composition or muscle strength. PMID:23201768

Hida, Azumi; Hasegawa, Yuko; Mekata, Yuko; Usuda, Mika; Masuda, Yasunobu; Kawano, Hitoshi; Kawano, Yukari

2012-01-01

178

Chromatographic adsorption of serum albumin and antibody proteins in cryogels with benzyl-quaternary amine ligands.  

PubMed

The preparation and characterization of mixed-mode adsorbents for a typical separation purpose are of great importance in bioseparation areas. In this work, we prepared a new monolithic cryogel with a combination of ion-exchange and hydrophobic functions by employing benzyl-quaternary amine groups. The fundamental cryogel properties, protein equilibrium adsorption isotherm and chromatographic adsorption in the cryogel were measured experimentally. The results showed that, by using bovine serum album as the model protein, the dual functional cryogel has protein binding capability even in salt solution and the buffer with pH close or below the protein isoelectric point due to both the electrostatic and hydrophobic interactions. A capillary-based adsorption model was developed, which provided satisfied insights of the microstructure, axial dispersion, mass transfer as well as protein adsorption characteristics within the cryogel bed. The chromatographic isolation of bioactive proteins from rabbit blood serum was carried out by the cryogel. Immunoglobulin G antibody with a purity of 98.2% and albumin with a purity of 96.8% were obtained, indicating that the cryogel could be an interesting and promising adsorbent in bioseparation areas. PMID:25618356

Yun, Junxian; Cheng, Xiuhong; Ye, Jialei; Shen, Shaochuan; Yang, Gensheng; Yao, Kejian; Kirsebom, Harald; Lin, Dong-Qiang; Guan, Yi-Xin; Yao, Shan-Jing

2015-02-13

179

Effect of bleaching permeate from microfiltered skim milk on 80% serum protein concentrate.  

PubMed

Whey proteins that have been removed before the cheese-making process are referred to as "native" whey proteins or milk serum proteins. Because serum proteins isolated directly from milk are not exposed to the cheese-making process, they are free from functional or sensory effects arising from this process. Whey proteins used in food and beverage applications are largely derived from annatto-colored Cheddar cheese. Some of the annatto is left in the whey and this color is converted to a colorless compound by bleaching. The effect of bleaching serum proteins on flavor and functionality of spray-dried protein provides a platform to investigate the effect of bleaching free from the confounding effects of cheese manufacture. The objective of this study was to characterize and compare the sensory and functional properties of 80% milk serum protein concentrate (SPC80) produced from bleached and unbleached microfiltration (MF) permeate made from skim milk with and without added annatto color. Colored and uncolored MF permeates were bleached with benzoyl peroxide (BP) or hydrogen peroxide (HP), ultrafiltered, diafiltered, and spray-dried. The SPC80 from unbleached colored and uncolored MF permeates were manufactured as controls. All treatments were manufactured in triplicate. All SPC80 were evaluated by sensory testing, instrumental analyses, functionality, color, and proximate analysis. The HP-bleached SPC80 was higher in lipid oxidation compounds than BP-bleached or unbleached SPC80, specifically hexanal, heptanal, nonanal, decanal, and 2,3-octadienone. The HP treatments were higher in aroma intensity and cardboard and fatty flavors compared with the unbleached and BP-bleached SPC80. The SPC80 bleached with BP had lower concentrations of norbixin compared with SPC80 bleached with HP. Functionality testing demonstrated that HP treatments had more soluble protein after 10min of heating at 90°C and pH 4.6 and pH 7 compared with the no bleach and BP treatments, regardless of additional color. Foams generated from bleached SPC80 were more stable than those from unbleached SPC80, and those bleached with HP were lower in yield stress than other SPC80. Overall, HP bleaching destroyed less norbixin and caused more lipid oxidation and subsequent off-flavors than did BP bleaching. However, the heat stability of SPC80 was enhanced by HP bleaching compared with control treatments or BP bleaching. PMID:23295111

Campbell, Rachel E; Adams, Michael C; Drake, Maryanne; Barbano, David M

2013-03-01

180

Serum heat shock protein 47 levels in patients with drug-induced lung disease  

PubMed Central

Background Heat shock protein (HSP) 47 is a collagen-specific molecular chaperone that is required for molecular maturation of various types of collagens. We recently reported that HSP47 serum levels were markedly higher in patients with acute exacerbations of idiopathic pulmonary fibrosis (IPF) when compared with patients with stable IPF, suggesting that serum HSP47 levels correlate with interstitial pneumonia activity. The aim of this study was to evaluate serum HSP47 levels in patients with drug-induced lung disease (DILD). Methods Findings from high-resolution computed tomographic chest scans of 47 patients with DILD were classified into one of four predominant patterns: organizing pneumonia (OP) (n?=?4), nonspecific interstitial pneumonia (NSIP) (n?=?24), hypersensitivity pneumonitis (HP) (n?=?11), and diffuse alveolar damage (DAD) (n?=?8). Serum levels of HSP47, Krebs von den Lungen-6 (KL-6), surfactant protein (SP)-A, and SP-D were measured in these patients. Results The PaO2/fraction of inspired oxygen (FiO2) (P/F) ratios were significantly lower and the alveolar-arterial difference of oxygen (A-a DO2) was significantly higher in the DAD group than in the other groups. Patients with DAD had the worst outcomes among the different subgroups. Patients in the DAD group had significantly higher serum HSP47 levels than those in other groups. Receiver operating characteristic curves revealed that HSP47 was superior to KL-6, SP-A, and SP-D for discriminating between the DAD group and the other groups. The cut-off level for HSP47 that resulted in the highest diagnostic accuracy was 1711.5 pg/mL. The sensitivity, specificity, and diagnostic accuracy were 87.5%, 97.4%, and 95.7%, respectively. Serum levels of HSP47 in the group of patients requiring glucocorticoids were significantly higher than those in patients who experienced clinical improvement without glucocorticoid administration. Serum HSP47 levels also significantly correlated with various respiratory parameters. Conclusion This study demonstrated that serum HSP47 levels were elevated in patients with DILD with a DAD pattern who had the worst outcomes among the different subgroups, and that this was correlated with P/F ratio and A-a DO2. PMID:24256690

2013-01-01

181

Serum protein inhibition of thyrotropin binding to human thyroid tissue. [¹²⁵I tracer technique  

Microsoft Academic Search

We used a modificaton of the TSH radioreceptor assay to detect TSH-binding inhibition (TBI) activity in serum and serum fractions from normal subjects and patients with Graves' disease. TBI activity is present in normal IgG prepared by DEAE-Sephadex chromatography and in normal globulins prepared by precipitation at 1.6 M ammonium sulfate. Other normal serum proteins also had TBI activity when

G. N. Beall; I. J. Chopra; D. H. Solomon; S. R. Kruger

1978-01-01

182

Long-term effects of radium exposure in female dial workers: Serum proteins  

Microsoft Academic Search

Serum protein levels were examined in 367 women first employed before 1930 in the radium dial industry. Mean levels were compared by age (55 to 64, 65 to 74, and 75 to 84 years) and initial intake of radium (<0.20, 0.20 to 0.99, greater than or equal to 1.00, and greater than or equal to 5.00 ..mu..Ci\\/kg). Few differences were

A. Polednak

1977-01-01

183

Comparison of inhibitory effects of oxygen radicals and calf serum protein on surfactant activity  

Microsoft Academic Search

The effects of the reactive oxygen species (ROS) superoxide anion (O\\u000a2\\u000a.\\u000a–) and hydroxyl radical (OH) on the surface tension lowering properties of bovine lipid extract surfactant (BLES) were compared to the effects of calf serum protein (CSP) in a captive bubble surfactometer (CBS). O\\u000a2\\u000a.\\u000a– was generated from xanthine\\/xanthine oxidase (X\\/XO), and OH was generated

M. M. Lee; F. H. Y. Green; S. Schürch; S. Cheng; S. G. Bjarnason; S. Leonard; W. Wallace; F. Possmayer; V. Vallyathan

2004-01-01

184

Quantification of Serum Proteins of Metastatic Oral Cancer Patients Using LC-MS/MS and iTRAQ Labeling  

PubMed Central

Metastasis is a critical event in oral squamous cell carcinoma (OSCC) progression. In this study, we have performed quantitative analysis of serum proteins from non-metastatic (lymph-node metastasis free) and metastatic OSCC patients using liquid chromatography-tandem mass spectrometry (LC-MS/MS) with iTRAQ labeling (isobaric tagging for relative and absolute quantitation). To eliminate highly abundant proteins, the serum samples were initially separated by SDS-PAGE and only low abundant protein bands were excised for subsequent in-gel tryptic digestion. The resulting peptides were then extracted from each sample gels and labeled with iTRAQ reagent 114 (control), 116 (non-metastatic) and 117 (metastatic), respectively. Afterwards, the labeled samples were combined and subjected to LC-MS/MS analysis using linear ion trap (LIT) MS with pulsed Q collision induced dissociation (PQD). A total of 64 proteins were identified and quantified by this approach. Our study showed that iTRAQ labeling and LIT-MS with PQD is a valuable approach to quantification of serum proteins. We also demonstrated the presence of differentially expressed serum proteins between non-metastatic and metastatic OSCCs that may be further validated as biomarkers for metastatic OSCC. However, in order to comprehensively quantify low abundant serum proteins, a more efficient approach is needed to deplete highly abundant proteins prior to quantitative serum proteome analysis of OSCC. PMID:20485468

Zhang, Lifeng; Jiang, Jiang; Arellano, Martha; Zhang, Lei; Yan, Xinmin; Wong, David T.; Hu, Shen

2010-01-01

185

The cell-shape protein MreC interacts with extracytoplasmic proteins including cell wall  

E-print Network

organization of cell wall (peptidoglycan) synthesizing complexes of penicillin- binding proteins (PBPs). Here as other proteins that lie outside the cytoplasmic membrane. MreB penicillin binding proteins peptidoglycan precursors by a family of enzymes known collectively as penicillin-binding proteins (PBPs) (1, 2). In general

Koehler, Carla

186

The presence of a novel protein in calf serum that recognizes beta amyloid in the formalin-fixed section.  

PubMed Central

Here we report on a monoclonal antibody, H6-33, that labels various beta-amyloid plaques, including diffuse plaques in the formalin-fixed, paraffin-embedded section from the brain affected with Alzheimer's disease (AD), without formic acid pretreatment. H6-33 also labels some neurofibrillary tangles and all kuru plaques in Gerstmann-Sträussler-Scheinker disease. In sharp contrast, H6-33 did not stain beta amyloid in the leptomeningeal vessel. For specific staining, H6-33 required the presence of fetal calf serum and it was necessary for beta amyloid to be formalin fixed. These results suggest that a novel protein in the calf serum, CSX, binds formalin-fixed beta amyloid, followed by H6-33 binding. The detection of beta amyloid by CSX was nullified by formic acid pretreatment of the tissue section. In accordance with this, CSX reacted only with a polymer form of synthetic beta peptide after fixation, but not with native beta-protein or beta-peptide monomer. These observations strongly suggest that 1) meningovascular beta amyloid should have a beta-pleated sheet structure somewhat dissimilar to that of beta-amyloid cores; and 2) most, if not all, of beta-protein immunoreactivities of diffuse plaques in AD sections are presumably derived from small amounts of amyloid fibrils scattered in the normal-looking neurohil. Images Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 PMID:1698030

Kanemaru, K.; Hasegawa, M.; Shimada, H.; Ihara, Y.

1990-01-01

187

Effects of Medroxyprogesterone Acetate and Caloric Restriction on the Distribution of Serum and Liver Lipids and Proteins in Female Rats  

Microsoft Academic Search

The effects of a large dose (2 mg\\/100 g body weight) of me- droxyprogesterone acetate (MPA), restricted feeding (50% of normal intake) and their interaction were investigated on the serum and liver lipid and protein distribution in female rats. MPA increased serum cholesterol and triglycéride levels in rats on ad libitum food intake without having any effect in the animals

PURABI DUTTA; RANDALL D. SMITH; MARGARET A. FLYNN

2010-01-01

188

Serum concentrations of cartilage oligomeric matrix protein and bone sialoprotein in hip osteoarthritis: A one year prospective study  

Microsoft Academic Search

OBJECTIVETo evaluate serum concentations of cartilage oligomeric matrix protein (COMP) and bone sialoprotein (BSP) as predictors of disease progression in hip osteoarthrtitis (OA).METHODSForty eight consecutive patients, referred to hospital for symptomatic hip OA, (ACR criteria) were monitored in a one year prospective trial with radiographs and serum samples. The radiographs were graded for joint space narrowing, osteophytes, and sclerosis and

Thierry Conrozier; Tore Saxne; Charles Shan Sei Fan; Pierre Mathieu; Anne-Marie Tron; Dick Heinegård; Eric Vignon

1998-01-01

189

Serum fatty acid binding protein 4, free fatty acids and metabolic risk markers  

PubMed Central

Fatty acid binding protein (FABP) 4 chaperones free fatty acids (FFA) in the adipocytes during lipolysis. Serum FFA relates to Metabolic Syndrome (METS) and serum FABP4 is emerging as a novel risk marker. In 36 overweight/obese women, serum FABP4 and FFA were measured hourly during 5-hour oral glucose tolerance test (OGTT). Insulin resistance was determined using frequently sampled intravenous GTT (FS-IVGTT). Serum lipids and inflammation markers were measured at fasting. During OGTT, serum FABP4 decreased by 40%, reaching its nadir at 3h (from 45.3±3.1 to 31.9±1.6 ng/mL) and stayed below the baseline at 5 h (35.9±2.2 ng/mL) (p < 0.0001 for both, compared to the baseline). Serum FFA decreased by 10 fold, reaching a nadir at 2h (from 0.611±0.033 to 0.067±0.004 mmol/L), then rebounded to 0.816±0.035 mmol/ L at 5h (p < 0.001 for both, compared to baseline). Both fasting-FABP4 and nadir-FABP4 correlated with obesity. Nadir-FABP4 correlated also with insulin resistance parameters from FS-IVGTT and with inflammation. Nadir-FFA, but not fasting-FFA, correlated with the METS-parameters. In conclusion, fasting-FABP4 related to metabolic risk markers more strongly than fasting-FFA. Nadir-FABP4 and nadir-FFA measured after glucose loading may provide better risk assessment than the fasting values. PMID:19394980

Karakas, Sidika E.; Almario, Rogelio U.; Kim, Kyoungmi

2009-01-01

190

Structural modification of serum vitamin D3-binding protein and immunosuppression in AIDS patients.  

PubMed

A serum glycoprotein, vitamin D3-binding protein (Gc protein), can be converted by beta-galactosidase of stimulated B lymphocytes and sialidase of T lymphocytes to a potent macrophage-activating factor (MAF), a protein with N-acetylgalactosamine as the remaining sugar moiety. Thus, Gc protein is a precursor for MAF. Treatment of purified Gc protein with immobilized beta-galactosidase and sialidase generates an extremely high-titered MAF (GcMAF). When peripheral blood monocytes/macrophages of 46 HIV-infected patients were treated with GcMAF (100 pg/ml), the monocytes/macrophages of all patients were efficiently activated. However, the MAF precursor activity of plasma Gc protein was low in 16 (35%) of of these patients. Loss of the MAF precursor activity appeared to be due to deglycosylation of plasma Gc protein by alpha-N-acetylgalactosaminidase found in the patient blood stream. Levels of plasma alpha-N-acetylgalactosaminidase activity in individual patients had an inverse correlation with the MAF precursor activity of their plasma Gc protein. Thus, precursor activity of Gc protein and alpha-N-acetylgalactosaminidase activity in patient blood can serve as diagnostic and prognostic indices. PMID:8573395

Yamamoto, N; Naraparaju, V R; Srinivasula, S M

1995-11-01

191

p53 protein, EGF receptor, and anti-p53 antibodies in serum from patients with occupationally derived lung cancer  

PubMed Central

The oncogene product epidermal growth factor receptor (EGF-R), the tumour suppressor gene product p53 and anti-p53 antibodies are detectable in the serum of certain cancer patients. Increased levels of some of these products were reported in lung cancer patients after occupational asbestos exposure and after exposure to polycyclic aromatic hydrocarbons or vinylchloride. In the first step, this study investigated the possible diagnostic value of serum EGF-R, p53-protein and anti-p53 antibodies, measured by an enzyme-linked immunosorbent assay, in lung tumour patients. In addition to being investigated on a molecular epidemiological basis, these parameters were examined as biomarkers of carcinogenesis, especially with regard to asbestos incorporation effects or of radon-induced lung cancers. Also, a possible effect of cigarette smoking and age dependence were studied. A total of 116 male patients with lung or pleural tumours were examined. The histological classification was four small-cell cancers, six large-cell cancers, 32 adenocarcinomas, 47 squamous carcinomas, 12 mixed lung carcinomas, five diffuse malignant mesotheliomas and ten lung metastasis of extrapulmonary tumours. Twenty-two lung cancers and all mesotheliomas were related to asbestos, 22 lung cancers were related to ionizing radiation and 61 patients had cigarette smoke-related lung cancer. Besides these patients 50 male patients with non-malignant lung or pleural diseases were included; of the latter eight subjects suffered from asbestosis. Controls were 129 male subjects without any lung disease. No significantly elevated or decreased serum values for p53 protein, EGF-R, or anti-p53 antibodies as a function of histological tumour type, age, or degree and type of exposure (asbestos, smoking, ionizing radiation) could be found. The utility of p53-protein, EGF-R and anti-p53 antibodies as routine biomarkers for screening occupationally derived lung cancers is limited. © 1999 Cancer Research Campaign PMID:10471051

Schneider, J; Presek, P; Braun, A; Bauer, P; Konietzko, N; Wiesner, B; Woitowitz, H-J

1999-01-01

192

THE INFLUENCE OF SERUM BINDING PROTEINS AND FEEDBACK CONTROL OF SERUM ESTRADIOL LEVELS ON THE COMPARATIVE POTENCY OF ENDOCRINE ACTIVE COMPOUNDS  

EPA Science Inventory

THE INFLUENCE OF SERUM BINDING PROTEINS ON THE COMPARATIVE RECEPTOR BINDING POTENCY OF ENDOCRINE ACTIVE COMPOUNDS. JG Teeguarden1 and HA Barton2. 1ICF Consulting, Research Triangle Park NC; 2US EPA, ORD, NHEERL, ETD, Pharmacokinetics Branch, RTP, NC. Accurate comparison of...

193

Increased serum levels of advanced oxidation protein products and glycation end products in subjects exposed to low-dose benzene.  

PubMed

Simple aromatic hydrocarbon benzene occurs naturally in crude oil and petroleum. Benzene has been internationally recognised as a haematotoxin and carcinogen. The involvement of oxidative stress is a major susceptibility factors for benzene hematotoxicity in humans. Advanced oxidation protein products (AOPPs) and advanced glycation end products (AGEs) are modified structures which can serve as markers of oxidative stress. The aim of this study is to assess modification of circulating AOPPs and AGEs, as early markers of oxidative stress, in subjects exposed to low dose of benzene. Furthermore the genetic polymorphism of glutathione-S-transferase (GST) may be related to health effects of benzene exposure, in fact both genotype T1 (GSTT1) and M1 (GSTM1) are involved in the detoxification of benzene oxide. The study was performed on 54 workers oil refinery employees. A group of 32 healthy age-matched subjects was included as controls. The AOPPs serum levels in oil refinery employees were higher in a statistically significant way than those measured in controls, but there were no significant changes in serum AGE levels between both groups. However, GST polymorphisms had not influence on serum levels of both biomarkers, so demonstrating that production of circulating AGEs and AOPPs in benzene parity-exposed workers levels is not dependent by GST genotypes. We can conclude that, in this condition, AOPPs are more sensitive marker of low benzene exposure than AGEs. PMID:22153878

Spatari, Giovanna; Saitta, Salvatore; Cimino, Francesco; Sapienza, Daniela; Quattrocchi, Paolina; Carrieri, Mariella; Barbaro, Mario; Saija, Antonella; Gangemi, Sebastiano

2012-04-01

194

Identification of serum monocyte chemoattractant protein-1 and prolactin as potential tumor markers in hepatocellular carcinoma.  

PubMed

Early diagnosis of hepatocellullar carcinoma (HCC) remains a challenge. The current practice of serum alpha-fetoprotein (AFP) measurement is inadequate. Here we utilized a proteomic approach to identify novel serum biomarkers for distinguishing HCC patients from non-cancer controls. We profiled the serum proteins in a group of 58 resectable HCC patients and 11 non-HCC chronic hepatitis B (HBV) carrier samples from the Singapore General Hospital (SGH) using the RayBio® L-Series 507 Antibody Array and found 113 serum markers that were significantly modulated between HCC and control groups. Selected potential biomarkers from this list were quantified using a multiplex sandwich enzyme-linked immunosorbent assay (ELISA) array in an expanded SGH cohort (126 resectable HCC patients and 115 non-HCC chronic HBV carriers (NC group)), confirming that serum prolactin and monocyte chemoattractant protein-1 (MCP-1) were significantly upregulated in HCC patients. This finding of serum MCP-1 elevation in HCC patients was validated in a separate cohort of serum samples from the Mochtar Riady Institute for Nanotechnology, Indonesia (98 resectable HCC, 101 chronic hepatitis B patients and 100 asymptomatic HBV/HCV carriers) by sandwich ELISA. MCP-1 and prolactin levels were found to correlate with AFP, while MCP-1 also correlated with disease stage. Subsequent receiver operating characteristic (ROC) analysis of AFP, prolactin and MCP-1 in the SGH cohort and comparing their area under the ROC curve (AUC) indicated that neither prolactin nor MCP-1 on their own performed better than AFP. However, the combination of AFP+MCP-1 (AUC, 0.974) had significantly superior discriminative ability than AFP alone (AUC, 0.942; p<0.001). In conclusion, prolactin and MCP-1 are overexpressed in HCC and are conveniently quantifiable in patients' sera by ELISA. MCP-1 appears to be a promising complementary biomarker for HCC diagnosis and this MCP-1+AFP model should be further evaluated as potential biomarker on a larger scale in patients at-risk of HCC. PMID:23874805

Wang, Who-Whong; Ang, Soo Fan; Kumar, Rajneesh; Heah, Charmain; Utama, Andi; Tania, Navessa Padma; Li, Huihua; Tan, Sze Huey; Poo, Desmond; Choo, Su Pin; Chow, Wan Cheng; Tan, Chee Kiat; Toh, Han Chong

2013-01-01

195

Serum heat shock protein 47 levels are elevated in acute interstitial pneumonia  

PubMed Central

Background Heat shock protein (HSP) 47, a collagen-specific molecular chaperone, is involved in the processing and/or secretion of procollagen. We hypothesized that HSP47 could be a useful marker for fibrotic lung disease. The aim of this study was to evaluate serum levels of HSP47 in patients with various idiopathic interstitial pneumonias (IIPs). Methods Subjects comprised 9 patients with acute interstitial pneumonia (AIP), 12 with cryptogenic organizing pneumonia (COP), 16 with nonspecific interstitial pneumonia (NSIP), 19 with idiopathic pulmonary fibrosis (IPF), and 19 healthy adult volunteers. Results Patients with AIP had serum HSP47 levels that were significantly higher than those of COP, NSIP or IPF patients and those of healthy volunteers. In contrast, serum levels of HSP47 among patients with COP, NSIP, IPF, and healthy volunteers did not differ significantly. Receiver operating characteristic curves revealed that the cut-off level for HSP47 that resulted in the highest diagnostic accuracy for discriminating between AIP and COP, NSIP, IPF, and healthy controls was 859.3 pg/mL. The sensitivity, specificity, and diagnostic accuracy were 100.0%, 98.5%, and 98.7%, respectively. Conclusion The present results demonstrate that, among patients with various IIPs, serum levels of HSP47 were elevated specifically in patients with AIP. PMID:24650086

2014-01-01

196

Estrogen Downregulation of Albumin and a 170-kDa Serum Protein in the Turtle, Trachemys scripta  

Microsoft Academic Search

We examined changes in serum protein composition after estradiol-17? treatment of ovariectomized female Trachemys scripta, with the objective of identifying proteins that are repressed by estrogen. The experimental protocol was validated by measuring serum estradiol-17? levels with a specific radioimmunoassay. Control turtle sera contained little or no estradiol-17? (mean = 25.8 pg\\/ml) while estrogen-treated turtle sera had elevated estradiol-17? levels

Kyle W. Selcer; Brent D. Palmer

1995-01-01

197

Deglycosylation of serum vitamin D3-binding protein leads to immunosuppression in cancer patients.  

PubMed

Serum vitamin D3-binding protein (Gc protein) can be converted by beta-galactosidase of B cells and sialidase of T cells to a potent macrophage activating factor, a protein with N-acetylgalactosamine as the remaining sugar moiety. Thus, Gc protein is the precursor of the macrophage activating factor (MAF). Treatment of Gc protein with immobilized beta-galactosidase and sialidase generates an extremely high titered MAF, Gc-MAF. When peripheral blood monocytes/macrophages of 52 patients bearing various types of cancer were incubated with 100 pg/ml of GcMAF, the monocytes/macrophages of all patients were efficiently activated. However, the MAF precursor activity of patient plasma Gc protein was found to be severely reduced in about 25% of this patient population. About 45% of the patients had moderately reduced MAF precursor activities. Loss of the precursor activity was found to be due to deglycosylation of plasma Gc protein by alpha-N-acetylgalactosaminidase detected in the patient's bloodstream. The source of the enzyme appeared to be cancerous cells. Radiation therapy decreased plasma alpha-N-acetylgalactosaminidase activity with concomitant increase of precursor activity. This implies that radiation therapy decreases the number of cancerous cells capable of secreting alpha-N-acetylgalactosaminidase. Both alpha-N-acetylgalactosaminidase activity and MAF precursor activity of Gc protein in patient bloodstream can serve as diagnostic and prognostic indices. PMID:8665521

Yamamoto, N; Naraparaju, V R; Asbell, S O

1996-06-15

198

[The immunogenic properties of the endotoxin protein: serum antibodies in animals and man].  

PubMed

Endotoxin protein or lipid A-associated protein (LAP) from Shigella sonnei was isolated and characterized earlier (Zh. mikrobiol. epidemiol. immunobiol., 1991, No. 4, pp. 47-50). In this investigation serum antibodies against LAP were studied in ELISA Anti-LAP antibodies were detected in high titers in the sera of nonimmunized mice, guinea pigs, rabbits, monkeys and healthy adults. We suppose that normal anti-LAP antibodies resulted from interaction between the immune system and environmental endotoxin. Parenteral injections of LAP to different animals induced intensive antibody response with a 100- to 1000-fold increase in the serum anti-LAP antibody level and a significant rise in the serum O-antibody level. The latter is seemingly due to the contamination of LAP with minute amounts of O-antigen (0.12% or less) and to the amplification of its immunogenicity by LAP. Both antigenic and amplifying activity of LAP was destroyed by proteinase K. The biological function of LAP and its possible use as a component of bacterial vaccines are briefly discussed. PMID:1719717

Belkin, Z P; Egorova, T P; Nartikova, V F; Fedosova, V G; Levenson, V I

1991-07-01

199

Biophysical analysis of the interaction of the serum protein human ?2GPI with bacterial lipopolysaccharide  

PubMed Central

There are several human serum proteins for which no clear role is yet known. Among these is the abundant serum protein beta2-glycoprotein-I (?2GPI), which is known to bind to negatively charged phospholipids as well as to bacterial lipopolysaccharides (LPS), and was therefore proposed to play a role in the immune response. To understand the details of these interactions, a biophysical analysis of the binding of ?2GPI to LPS and phosphatidylserine (PS) was performed. The data indicate only a moderate tendency of the protein (1) to influence the LPS-induced cytokine production in vitro, (2) to react exothermally with LPS in a non-saturable way, and (3) to change its local microenvironment upon LPS association. Additionally, we found that the protein binds more strongly to phosphatidylserine (PS) than to LPS. Furthermore, ?2GPI converts the LPS bilayer aggregates into a stronger multilamellar form, and reduces the fluidity of the hydrocarbon moiety of LPS due to a rigidification of the acyl chains. From these data it can be concluded that ?2GPI plays a role as an immune-modulating agent, but there is much less evidence for a role in immune defense against bacterial toxins such as LPS. PMID:24918058

Gries, Anna; Prassl, Ruth; Fukuoka, Satoshi; Rössle, Manfred; Kaconis, Yani; Heinbockel, Lena; Gutsmann, Thomas; Brandenburg, Klaus

2014-01-01

200

Pattern of serum protein fractions in dairy cows during different stages of gestation and lactation.  

PubMed

In dairy cows the period of transition from late gestation to early lactation is recognized as inducing considerable metabolic adaptation. The aim of this study was to analyse modifications in serum protein values occurring during the dry and the transition period and during lactation in a group of five Holstein cows of high average milk production. For all subjects, selected on the basis of their pregnancy status, blood samples were collected at different physiological phases: dry period (-60, -30 d to calving), transition period (almost 7 d to calving, 7 d after calving), and lactation (weeks 2, 5 and 15 after calving), for a total of eight blood samples for each cow. On each blood sample total proteins and electrophoresis analysis were performed. On the data obtained, normally distributed (P<0·05, Kolmogorov-Smirnov's test), one-way Repeated Measure Analysis of Variance (ANOVA), was applied to evaluate the influence of different stages of gestation and lactation on the considered parameters. Results showed a significant effect on total proteins, ?1-globulins, ?-globulins, ?-globulins and albumin/globulin ratio. Most of the detected modifications were related to the transition from gestation to lactation, indicating that it is a period of great metabolic stress for cows. On the basis of the obtained results we can affirm that the pattern of serum protein fraction rn could give information about dehydration, plasma volume expansion and hepatic function occurring during the peripartum period in dairy cows. PMID:21843395

Piccione, Giuseppe; Messina, Vanessa; Schembari, Angelo; Casella, Stefania; Giannetto, Claudia; Alberghina, Daniela

2011-11-01

201

The Association between Preoperative Serum C-Reactive Protein and Hepatocellular Carcinoma Recurrence in Patients with Chronic Hepatitis B Virus (HBV) Infection—A Retrospective Study  

PubMed Central

The prognosis of the patients with hepatocellular carcinoma (HCC) recurrence following curative hepatectomy is usually dismal. Whether preoperative serum C-reactive protein (CRP) can predict the recurrence of HCC in patients with chronic HBV infection is not clear. Total 232 patients with chronic HBV infection were included in this retrospective study. We investigated the association between detailed preoperative serum CRP levels and early (? 2 year) and late (> 2 year) HCC recurrence following curative hepatectomy. After adjusting for potential confounders, we found a saturation effect for preoperative serum CRP of 2.1 mg/dl existed for early HCC recurrence (ER). The incidence of ER increased with preoperative serum CRP less than 2.1 mg/dl (OR = 3.5, 95% CI 1.6–7.6, P = 0.001), and higher preoperative serum CRP (>2.1 mg/dl) did not increase the incidence of ER (OR = 0.8, 95% CI 0.2–2.7, P = 0.703). Whereas there is a linear relationship between preoperative serum CRP and late HCC recurrence (LR) (OR = 0.2, 95% CI, 0.1- 0.4) (OR = 1.8, 95% CI, 1.2–2.5, P = 0.002). In addition, the optimal cutoff point for serum CRP level was 1.5 mg/dl, instead of 1.0 mg/dl, in predicting both ER and LR. Patients with higher preoperative serum CRP level (>1.5 mg/dl) had lower recurrence free survival rates and overall survival rates (P<0.01). These results suggest that preoperative serum CRP played different roles on ER and LR following curative hepatectomy, thus further predictingthe prognosis in patients with chronic HBV infection. PMID:25602444

Zhao, Xiaoming; Luo, Jingyu; Li, Bobo; Liu, Shuguang; Li, Daotang

2015-01-01

202

Serum protein binding and the role of increased alpha 1-acid glycoprotein in moderately obese male subjects.  

PubMed Central

Serum protein and lipid concentrations as well as the serum protein binding of propranolol, diazepam and phenytoin were measured in normal weight and obese volunteers. Concentrations of alpha 1-acid glycoprotein (AAG) in the obese subjects were double that of the lean controls. Conversely, concentrations of high density lipoproteins (HDL) were decreased in the obese group. The serum binding of propranolol was increased in the obese subjects and correlated with serum AAG concentrations. Diazepam binding was slightly decreased in the obese as a result of lower serum albumin concentrations and elevated free fatty acids. The binding of phenytoin was comparable in all of the volunteers. These findings point out some of the complex pathophysiologic changes associated with obesity which may in turn influence drug disposition and hence drug therapy in the obese patient. PMID:6529534

Benedek, I H; Blouin, R A; McNamara, P J

1984-01-01

203

Immunochemical and ultrastructural study of multiple myeloma with a heavy chain protein in the serum.  

PubMed Central

A patient with multiple myeloma had antigenically related monoclonal Fc-gamma fragments and complete IgG-kappa molecules in the serum. The urine contained only Fc-gamma fragments in the absence of Bence-Jones protein. The two distinct M-components in the serum showed electrophoretic identity but could be separated by chromatography. The simultaneous presence of complete monoclonal IgG molecules and Fc-gamma fragments, though difficult to detect, could be a frequent occurrence in multiple myeloma, and it could be defined as 'double paraproteinaemia'. A detailed ultrastructural study was performed in this case and showed fibril bundles being released from the malignant plasma cells; such fibrils could be the supramolecular organisation of the neosynthesised heavy chain fragments. Images Fig. 10 Fig. 1 Fig. 4 Fig. 5 Fig. 6 Fig. 7 Fig. 8 Fig. 9 Fig. 11 PMID:6776150

Bartoloni, C; Flamini, G; Gentiloni, N; Russo, M A; Barone, C; Gambassi, G; Terranova, T

1980-01-01

204

Autocrine induction of collagenase by serum amyloid A-like and beta 2-microglobulin-like proteins.  

PubMed

Two autocrine proteins of 14 and 12 kilodaltons that induce the synthesis of rabbit fibroblast collagenase were identified. The proteins were purified from serum-free culture medium taken from rabbit synovial fibroblasts stimulated with phorbol myristate acetate. The amino-terminal sequences of the 14- and 12-kilodalton species were approximately 60 to 80 percent homologous with serum amyloid A and beta 2 microglobulin, respectively. The polyacrylamide gel-eluted proteins retained the ability to induce collagenase synthesis in rabbit and human fibroblasts. These autocrine proteins may provide a means to modulate collagenase synthesis in normal remodeling as well as in inflammation and disease states. PMID:2536953

Brinckerhoff, C E; Mitchell, T I; Karmilowicz, M J; Kluve-Beckerman, B; Benson, M D

1989-02-01

205

Quantitation of Serum Prostate-specific Membrane Antigen by a Novel Protein Biochip Immunoassay Discriminates Benign from Malignant Prostate Disease1  

Microsoft Academic Search

The lack of a sensitive immunoassay for quantitating serum prostate- specific membrane antigen (PSMA) hinders its clinical utility as a diagnostic\\/ prognostic biomarker. An innovative protein biochip immunoassay was used to quantitate and compare serum PSMA levels in healthy men and patients with either benign or malignant prostate disease. PSMA was captured from serum by anti-PSMA antibody bound to ProteinChip

Zhen Xiao; Bao-Ling Adam; Lisa H. Cazares; John W. Davis; Paul F. Schellhammer; Enrique A. Dalmasso; George L. Wright

206

Resilience to bacterial infection: difference between species could be due to proteins in serum  

PubMed Central

Vertebrates vary in resistance and resilience to infectious diseases, and the mechanisms regulating the trade-off between these two often opposing protective processes are not well understood. Variability in the sensitivity of species to induction of damaging inflammation in response to equivalent pathogen loads (resilience) complicates the use of animal models that reflect human disease. We found that induction of pro-inflammatory cytokines from macrophages in response to inflammatory stimuli in vitro is regulated by proteins in the sera of species in inverse proportion to their in vivo resilience to lethal doses of bacterial lipopolysaccharide over a range of 10,000-fold. This finding suggests that proteins in serum rather than intrinsic cellular differences may play a role in regulating variations in resilience to microbe-associated molecular patterns between species. Involvement of circulating proteins as key molecules raises hope that the process might be manipulated to create better animal models and potentially new drug targets. PMID:20001600

Warren, H. Shaw; Fitting, Catherine; Hoff, Eva; Adib-Conquy, Minou; Beasley-Topliffe, Laura; Tesini, Brenda; Liang, Xueya; Valentine, Catherine; Hellman, Judith; Hayden, Douglas; Cavaillon, Jean-Marc

2009-01-01

207

Combined evaluation of a panel of protein and miRNA serum-exosome biomarkers for pancreatic cancer diagnosis increases sensitivity and specificity.  

PubMed

Late diagnosis contributes to pancreatic cancer (PaCa) dismal prognosis, urging for reliable, early detection. Serum-exosome protein and/or miRNA markers might be suitable candidates, which we controlled for patients with PaCa. Protein markers were selected according to expression in exosomes of PaCa cell line culture supernatants, but not healthy donors' serum-exosomes. miRNA was selected according to abundant recovery in microarrays of patients with PaCa, but not healthy donors' serum-exosomes and exosome-depleted serum. According to these preselections, serum-exosomes were tested by flow cytometry for the PaCa-initiating cell (PaCIC) markers CD44v6, Tspan8, EpCAM, MET and CD104. Serum-exosomes and exosome-depleted serum was tested for miR-1246, miR-4644, miR-3976 and miR-4306 recovery by qRT-PCR. The majority (95%) of patients with PaCa (131) and patients with nonPa-malignancies reacted with a panel of anti-CD44v6, -Tspan8, -EpCAM and -CD104. Serum-exosomes of healthy donors' and patients with nonmalignant diseases were not reactive. Recovery was tumor grading and staging independent including early stages. The selected miR-1246, miR-4644, miR-3976 and miR-4306 were significantly upregulated in 83% of PaCa serum-exosomes, but rarely in control groups. These miRNA were also elevated in exosome-depleted serum of patients with PaCa, but at a low level. Concomitant evaluation of PaCIC and miRNA serum-exosome marker panels significantly improved sensitivity (1.00, CI: 0.95-1) with a specificity of 0.80 (CI: 0.67-0.90) for PaCa versus all others groups and of 0.93 (CI: 0.81-0.98) excluding nonPa-malignancies. Thus, the concomitant evaluation of PaCIC and PaCa-related miRNA marker panels awaits retrospective analyses of larger cohorts, as it should allow for a highly sensitive, minimally-invasive PaCa diagnostics. PMID:25388097

Madhavan, Bindhu; Yue, Shijing; Galli, Uwe; Rana, Sanyukta; Gross, Wolfgang; Müller, Miryam; Giese, Nathalia A; Kalthoff, Holger; Becker, Thomas; Büchler, Markus W; Zöller, Margot

2014-11-12

208

Identification of differential pattern of protein expression in canine osteoarthritis serum after anterior cruciate ligament transection: a proteomic analysis.  

PubMed

Osteoarthritis (OA) management remains a great challenge and there is considerable effort to understand its pathophysiology and to identify new therapeutic targets and biomarkers. Canine OA surgically induced by the transection of the anterior cruciate ligament (ACLT) is a widely used and relevant model. This study reports a proteome mapping of dog serum and an analysis of the differentially expressed proteins between before and after ACLT. In the first part of the study, 261 picked protein spots were identified from preparative 2D gels and 71 different proteins were identified among the 261 spots present on the reference map. Canine serum proteome mapping reveals the presence of proteins of interest, such as fetuin B, complement C3 and C1s and pregnancy zone protein. The comparison between serum from dogs before and after ACLT reveals the differential expression of several proteins that could play a key role in the pathogenesis of OA. A number of proteins, such as fetuin B and complement C3, were increased in dog OA serum whereas others, such as hyaluronan binding protein 2, inter-alpha-trypsin inhibitor H4 (ITIH4), complement C1s and C4 and haptoglobin were decreased. Some of these proteins could be candidate biomarkers for diagnosis, prognosis and treatment evaluation. The results of the study also reinforced the similarities between dog experimental OA and human cases of OA. PMID:23831215

Gharbi, M; Sanchez, C; Mazzucchelli, G; De Pauw, E; Henrotin, Y

2013-09-01

209

Serum Vitamin D, Vitamin D Binding Protein, and Risk of Colorectal Cancer  

PubMed Central

Background We previously reported a positive association between serum 25-hydroxyvitamin D (25(OH)D) and colorectal cancer risk. To further elucidate this association, we examined the molar ratio of 25(OH)D to vitamin D binding protein (DBP), the primary 25(OH)D transport protein, and whether DBP modified the association between 25(OH)D and colorectal cancer risk. Methods In a nested case-control study within the Alpha-Tocopherol, Beta-Carotene Cancer Prevention Study, controls were 1?1 matched to 416 colorectal cancer cases based on age and date of blood collection. Logistic regression was used to estimate odds ratios (ORs) and 95% confidence intervals (CI) for quartiles of 25(OH)D, DBP, and the molar ratio of 25(OH)D:DBP, a proxy for free, unbound circulating 25(OH)D. Results Comparing highest to lowest quartiles, DBP was not associated with colorectal cancer risk (OR?=?0.91; 95% CI: 0.58, 1.42, p for trend ?=?0.58); however, a positive risk association was observed for the molar ratio of 25(OH)D:DBP (OR?=?1.44; 95% CI: 0.92, 2.26, p for trend ?=?0.04). In stratified analyses, the positive association between 25(OH)D and colorectal cancer was stronger among men with DBP levels above the median (OR?=?1.89; 95% CI: 1.07, 3.36, p for trend ?=?0.01) than below the median (OR?=?1.20; 95% CI: 0.68, 2.12, p for trend ?=?0.87), although the interaction was not statistically significant (p for interaction ?=?0.24). Conclusion Circulating DBP may influence the association between 25(OH)D and colorectal cancer in male smokers, with the suggestion of a stronger positive association in men with higher DBP concentrations. This finding should be examined in other populations, especially those that include women and non-smokers. PMID:25036524

Anic, Gabriella M.; Weinstein, Stephanie J.; Mondul, Alison M.; Männistö, Satu; Albanes, Demetrius

2014-01-01

210

A study on blood groups and serum proteins in Bengalee populations of Calcutta, India.  

PubMed

The genetic polymorphisms of two blood groups (A1A2B0, Rh-D) and two serum proteins (HP, TF) were investigated in five endogamous caste groups of Bengalee Hindu population living in Calcutta. The distribution of A1A2B0 and Rh-D blood groups in all the caste groups showed an oriental pattern with high B and Rh-D+ frequencies, while for the haptoglobins a very low frequency of HP*1 was seen in all the caste groups, except the Vaidya. For transferrin types the absolute predominancy of TF*C in all the caste groups was noted. PMID:7993067

Bandyopadhyay, A R

1994-09-01

211

Interaction of lipid vesicle with silver nanoparticle-serum albumin protein corona  

NASA Astrophysics Data System (ADS)

The physical interaction between a lipid vesicle and a silver nanoparticle (AgNP)-human serum albumin (HSA) protein "corona" has been examined. Specifically, the binding of AgNPs and HSA was analyzed by spectrophotometry, and the induced conformational changes of the HSA were inferred from circular dichroism spectroscopy. The fluidity of the vesicle, a model system for mimicking cell membrane, was found to increase with the increased exposure to AgNP-HSA corona, though less pronounced compared to that induced by AgNPs alone. This study offers additional information for understanding the role of physical forces in nanoparticle-cell interaction and has implications for nanomedicine and nanotoxicology.

Chen, Ran; Choudhary, Poonam; Schurr, Ryan N.; Bhattacharya, Priyanka; Brown, Jared M.; Chun Ke, Pu

2012-01-01

212

Comparative serum proteomic analysis involving liver organ-specific metastasis-associated proteins of nasopharyngeal carcinoma  

PubMed Central

Metastasis is the main cause of cancer-related mortality; patients with liver metastases (LM) have the worst prognosis among patients with nasopharyngeal carcinoma (NPC). However, at present, few biomarkers for detecting organ-specific metastasis have been identified. Proteomics, an ultra-sensitive analytical technique, can detect molecular changes before organ-specific metastasis occurs. Analysis with matrix-assisted, laser desorption-ionization time-of-flight mass spectrometry (MALDI-TOF-MS), combined with magnetic chemical affinity beads is a new technique for evaluating protein separation. We sought to identify potential liver-specific, metastasis-associated proteomic printing in patients with NPC. We examined 64 serum samples from 50 patients who had pathologically confirmed NPC and 14 who had pathologically confirmed non-NPC with LM using MALDI-TOF-MS with weak cation bead protein chips. During follow-up of at least 37 months (maximum, 176 months) following radiotherapy, we confirmed 16 cases of LM (LM NPC), 16 cases without LM (non-LM NPC) and 18 cases without metastasis (non-M NPC). Using comparison analysis, 4 protein mass peaks, 4155.34, 4194.87, 4210.78 and 4249.56 m/z were identified as liver-specific, metastasis-associated protein peaks in NPC and two of them (4155 and 4249 m/z) met two different statistical criteria in both ClinProt software analyses and discriminant analyses. Models based on the 4 potential serum markers of NPC discriminated between LM NPC, non-LM NPC, non-M NPC and non-NPC LM analyzed with sieved markers. The recognition capability and cross-validation of these models for differentiating the above 4 groups are all approximately 80%. MALDI-TOF-MS combined with tree analysis models may provide a clinical diagnostic platform for detecting potential liver-specific, metastasis-associated proteomic printing in NPC. However, markedly differential proteins still need to be identified. PMID:22970016

PAN, CHANGCHUAN; TAO, YALAN; ZHAO, MING; LI, WANG; HUANG, ZILIN; GAO, JING; WU, YANHEN; YU, JINGRUI; WU, PEIHONG; XIA, YUNFEI; LU, JIN

2012-01-01

213

Serum-stable quantum dot--protein hybrid nanocapsules for optical bio-imaging  

NASA Astrophysics Data System (ADS)

We introduce shell cross-linked protein/quantum dot (QD) hybrid nanocapsules as a serum-stable systemic delivery nanocarrier for tumor-targeted in vivo bio-imaging applications. Highly luminescent, heavy-metal-free Cu0.3InS2/ZnS (CIS/ZnS) core-shell QDs are synthesized and mixed with amine-reactive six-armed poly(ethylene glycol) (PEG) in dichloromethane. Emulsification in an aqueous solution containing human serum albumin (HSA) results in shell cross-linked nanocapsules incorporating CIS/ZnS QDs, exhibiting high luminescence and excellent dispersion stability in a serum-containing medium. Folic acid is introduced as a tumor-targeting ligand. The feasibility of tumor-targeted in vivo bio-imaging is demonstrated by measuring the fluorescence intensity of several major organs and tumor tissue after an intravenous tail vein injection of the nanocapsules into nude mice. The cytotoxicity of the QD-loaded HSA-PEG nanocapsules is also examined in several types of cells. Our results show that the cellular uptake of the QDs is critical for cytotoxicity. Moreover, a significantly lower level of cell death is observed in the CIS/ZnS QDs compared to nanocapsules loaded with cadmium-based QDs. This study suggests that the systemic tumor targeting of heavy-metal-free QDs using shell cross-linked HSA-PEG hybrid nanocapsules is a promising route for in vivo tumor diagnosis with reduced non-specific toxicity.

Lee, Jeong Yu; Nam, Dong Heon; Oh, Mi Hwa; Kim, Youngsun; Choi, Hyung Seok; Jeon, Duk Young; Beum Park, Chan; Nam, Yoon Sung

2014-05-01

214

Association of serum amyloid A protein and peptide fragments with prognosis in renal cancer  

PubMed Central

Background: In renal cell carcinoma (RCC), the discovery of biomarkers for clinical use is a priority. This study aimed to identify and validate diagnostic and prognostic serum markers using proteomic profiling. Methods: Pre-operative sera from 119 patients with clear cell RCC and 69 healthy controls was analysed by surface-enhanced laser desorption/ionisation time-of-flight mass spectrometry with stringent in-house quality control and analysis routines. Following identification of one prognostic peak as a fragment of serum amyloid A (SAA), total serum SAA and CRP were also determined by immunoassay for further validation. Results: Several peptides were identified as having independent prognostic but not diagnostic significance on multivariable analysis. One was subsequently identified as a 1525?Da fragment of SAA (hazard ratio (HR)=0.26, 95% CI 0.08–0.85, P=0.026). This was weakly negatively correlated with total SAA, which was also of independent prognostic significance (HR=2.46, 95% CI 1.17–5.15, P=0.017). Both potentially strengthened prognostic models based solely on pre-operative variables. Conclusions: This is the first description of the prognostic value of this peptide in RCC and demonstrates proof of principle of the approach. The subsequent examination of SAA protein considerably extends previous studies, being the first study to focus solely on pre-operative samples and describing potential clinical utility in pre-operative prognostic models. PMID:20531413

Wood, S L; Rogers, M; Cairns, D A; Paul, A; Thompson, D; Vasudev, N S; Selby, P J; Banks, R E

2010-01-01

215

The effects of hemodialysis and peritoneal dialysis on serum homocysteine and C-reactive protein levels.  

PubMed Central

OBJECTIVES: In this study, we aimed to investigate plasma homocysteine (Hcy) and serum C-reactive protein (CRP) levels in hemodialysis (HD) and continuous ambulatory peritoneal dialysis (CAPD) patients, and the relation among them. MATERIALS AND METHODS: This study was carriedout on 52 HD patients, 26 CAPD patients and a control group of 22 healthy persons. Blood samples were taken from the patients for Hcy and CRP measurements. RESULTS: Serum CRP level was found to be high in 48.1% of HD patients, 69.2% of CAPD patients and 4.5% of the healthy control group. Plasma Hcy level was found out to be above the normal limits in 73.1% of HD patients, 65.4% of CAPD patients and 9% of the healthy control group. There was a significant positive relation (r = 0.384, p < 0.001) between the levels of plasma Hcy and serum CRP in HD and CAPD patients. CONCLUSION: The high levels of Hcy and CRP were found out to be higher in HD and CAPD patients than in the control group. In order to determine the risk rate of Hcy and CRP for coronary artery disease, extensive investigations are required in patients with chronic renal failure that also have coronary artery disease. PMID:15770053

Borazan, Ali; Aydemir, Selim; Sert, Mehmet; Yilmaz, Ahmet

2004-01-01

216

Comparison of serum amyloid A and C-reactive protein as diagnostic markers of systemic inflammation in dogs  

PubMed Central

The diagnostic performance of canine serum amyloid A (SAA) was compared with that of C-reactive protein (CRP) in the detection of systemic inflammation in dogs. Sera from 500 dogs were retrospectively included in the study. C-reactive protein and SAA were measured using validated automated assays. The overlap performance, clinical decision limits, overall diagnostic performance, correlations, and agreement in the clinical classification between these 2 diagnostic markers were compared. Significantly higher concentrations of both proteins were detected in dogs with systemic inflammation (SAA range: 48.75 to > 2700 mg/L; CRP range: 0.4 to 907.4 mg/L) compared to dogs without systemic inflammation (SAA range: 1.06 to 56.4 mg/L; CRP range: 0.07 to 24.7 mg/L). Both proteins were shown to be sensitive and specific markers of systemic inflammation in dogs. Significant correlations and excellent diagnostic agreement were observed between the 2 markers. However, SAA showed a wider range of concentrations and a significantly superior overall diagnostic performance compared with CRP. PMID:24489396

Christensen, Michelle B.; Langhorn, Rebecca; Goddard, Amelia; Andreasen, Eva B.; Moldal, Elena; Tvarijonaviciute, Asta; Kirpensteijn, Jolle; Jakobsen, Sabrina; Persson, Frida; Kjelgaard-Hansen, Mads

2014-01-01

217

Comparison of serum amyloid A and C-reactive protein as diagnostic markers of systemic inflammation in dogs.  

PubMed

The diagnostic performance of canine serum amyloid A (SAA) was compared with that of C-reactive protein (CRP) in the detection of systemic inflammation in dogs. Sera from 500 dogs were retrospectively included in the study. C-reactive protein and SAA were measured using validated automated assays. The overlap performance, clinical decision limits, overall diagnostic performance, correlations, and agreement in the clinical classification between these 2 diagnostic markers were compared. Significantly higher concentrations of both proteins were detected in dogs with systemic inflammation (SAA range: 48.75 to > 2700 mg/L; CRP range: 0.4 to 907.4 mg/L) compared to dogs without systemic inflammation (SAA range: 1.06 to 56.4 mg/L; CRP range: 0.07 to 24.7 mg/L). Both proteins were shown to be sensitive and specific markers of systemic inflammation in dogs. Significant correlations and excellent diagnostic agreement were observed between the 2 markers. However, SAA showed a wider range of concentrations and a significantly superior overall diagnostic performance compared with CRP. PMID:24489396

Christensen, Michelle B; Langhorn, Rebecca; Goddard, Amelia; Andreasen, Eva B; Moldal, Elena; Tvarijonaviciute, Asta; Kirpensteijn, Jolle; Jakobsen, Sabrina; Persson, Frida; Kjelgaard-Hansen, Mads

2014-02-01

218

Serum Albumin Prevents Protein Aggregation and Amyloid Formation and Retains Chaperone-like Activity in the Presence of Physiological Ligands  

PubMed Central

Although serum albumin has an established function as a transport protein, evidence is emerging that serum albumin may also have a role as a molecular chaperone. Using established techniques to characterize chaperone interactions, this study demonstrates that bovine serum albumin: 1) preferentially binds stressed over unstressed client proteins; 2) forms stable, soluble, high molecular weight complexes with stressed client proteins; 3) reduces the aggregation of client proteins when it is present at physiological levels; and 4) inhibits amyloid formation by both WT and L55P transthyretin. Although the antiaggregatory effect of serum albumin is maintained in the presence of physiological levels of Ca2+ and Cu2+, the presence of free fatty acids significantly alters this activity: stabilizing serum albumin at normal levels but diminishing chaperone-like activity at high concentrations. Moreover, here it is shown that depletion of albumin from human plasma leads to a significant increase in aggregation under physiologically relevant heat and shear stresses. This study demonstrates that serum albumin possesses chaperone-like properties and that this activity is maintained under a number of physiologically relevant conditions. PMID:22549788

Finn, Thomas E.; Nunez, Andrea C.; Sunde, Margaret; Easterbrook-Smith, Simon B.

2012-01-01

219

Methylation of serum insulin-like growth factor-binding protein 7 promoter in hepatitis B virus-associated hepatocellular carcinoma.  

PubMed

Methylation of gene promoter CpG islands is an important early event in hepatocellular carcinoma (HCC), and detection of cell-free tumor-specific DNA methylation is becoming a useful noninvasive method for HCC. This study was aimed at determining the diagnostic value of serum insulin-like growth factor-binding protein 7 (IGFBP7) promoter methylation in hepatitis B virus-associated HCC. A total of 217 subjects, including 136 HCC patients, 46 patients with chronic hepatitis B (CHB), and 35 healthy controls (HCs), were included. The methylation status of the serum IGFBP7 gene promoter was determined using methylation-specific PCR. The frequency of serum IGFBP7 promoter methylation in HCC patients (89/136, 65%) was significantly higher than that in CHB patients (8/46, 17%; X(2) = 31.883, P < 0.001) and HCs (5/35, 14%; X(2) = 29.429, P < 0.001). Moreover, elevated IGFBP7 methylation frequency was also observed in HCC patients with vascular invasion compared with those without vascular invasion (84 versus 60%, X(2) = 6.633, P = 0.010). The sensitivities of serum IGFBP7 methylation and alpha-fetoprotein (AFP) in detecting HCC were 65 and 57%, respectively. Of note, the combination of IGFBP7 methylation and AFP showed 85% for sensitivity. These results suggest that methylation of the serum IGFBP7 gene promoter may serve as a useful noninvasive biomarker for HCC diagnosis. PMID:24142767

Li, Feng; Fan, Yu-Chen; Gao, Shuai; Sun, Feng-Kai; Yang, Yang; Wang, Kai

2014-01-01

220

Correlation between Ocular Demodex Infestation and Serum Immunoreactivity to Bacillus Proteins in Patients with Facial Rosacea  

PubMed Central

Purpose To investigate correlation between ocular Demodex infestation and serum. Design A prospective study to correlate clinical findings with laboratory data. Participants We consecutively enrolled 59 patients: 34 men and 25 women with a mean age of 60.4±17.6 years (range, 17–93). Methods Demodex counting was performed based on lash sampling. Serum immunoreactivity to two 62-kDa and 83-kDa proteins derived from B oleronius was determined by Western blot analysis. Facial rosacea, lid margin, and ocular surface inflammation were documented by photography and graded in a masked fashion. Main Outcome Measures Statistical significance based on correlative analyses of clinical and laboratory data. Results These 59 patients were age matched, but not gender matched, regarding serum immunoreactivity, ocular Demodex infestation, or facial rosacea. There was a significant correlation between serum immunoreactivity and facial rosacea (P = 0.009), lid margin inflammation (P = 0.040), and ocular Demodex infestation (P = 0.048), but not inferior bulbar conjunctival inflammation (P = 0.573). The Demodex count was significantly higher in patients with positive facial rosacea (6.6±9.0 vs. 1.9±2.2; P = 0.014). There was a significant correlation of facial rosacea with lid margin inflammation (P = 0.016), but not with inferior bulbar conjunctival inflammation (P = 0.728). Ocular Demodex infestation was less prevalent in patients with aqueous tear-deficiency dry eye than those without (7/38 vs. 12/21; P = 0.002). Conclusions The strong correlation provides a better understanding of comorbidity between Demodex mites and their symbiotic B oleronius in facial rosacea and blepharitis. Treatments directed to both warrant future investigation. PMID:20079929

Li, Jianjing; O'Reilly, Niamh; Sheha, Hosam; Katz, Raananah; Raju, Vadrevu K.; Kavanagh, Kevin; Tseng, Scheffer C. G.

2010-01-01

221

Structural insights into the binding of uranyl with human serum protein apotransferrin structure and spectra of protein-uranyl interactions.  

PubMed

Ab initio quantum mechanical computational studies for the structure and IR spectra of the uranyl complex with human serum apotransferrin (TF) protein are carried out to model uranyl intake into the human cell through endocytosis and formation of a coordination complex with the protein binding sites. The computed IR spectra and structure of the uranyl-protein complex facilitate interpretation of the observed spectra and confirm the primary binding sites of the transferrin protein with the uranyl ion. Our computed equilibrium geometry and the IR spectra of the uranyl-TF complex reveal that uranyl ion is bound to two tyrosines, one aspartate group, and one carbonate ion. Our IR spectra indicate that histidine is not involved in binding to uranyl with transferrin protein. Our computations reveal a short, strong hydrogen bond, which could play an important role in the stabilization and formation of the uranyl-TF complex. Computed Laplacian charge plots indicate high chemical reactivity on this complex as both an electrophile and a nucleophile, facilitating binding to different receptors and thus entry into a number of target organs and the blood-brain barrier. The Mulliken charge density plots and the three-dimensional charge density plots suggest a donor-acceptor mechanism in the complex formation. PMID:19678663

Benavides-Garcia, Maria G; Balasubramanian, Krishnan

2009-09-01

222

Identification of a novel serum amyloid A protein in BALB/c mice.  

PubMed Central

Four serum amyloid A protein (SAA) genes and two SAA gene products, SAA1 and SAA2, were identified in BALB/c mice. Using analytical isoelectric focusing we have identified a quantitatively significant new member of the SAA family and designated it 'SAA5'. This protein has characteristics never before described for any SAA molecule. In the highly conserved region between amino acids 33 and 44, identical in all SAAs from all species examined, SAA5 had four amino acid substitutions. In addition, the induction of SAA5 by lipopolysaccharide had different kinetics from that of the other mouse SAAs. Our data suggest that the mouse SAA gene family is more complex in composition and regulation than previously surmised. Images Fig. 1. Fig. 2. Fig. 3. PMID:1741755

de Beer, M C; Beach, C M; Shedlofsky, S I; de Beer, F C

1991-01-01

223

Interaction between rat serum phosphorylcholine binding protein and platelet activating factor.  

PubMed

The binding of rat serum phosphorylcholine binding protein (PCBP) to platelet activating factor (PAF) has been demonstrated using a HPLC-gel filtration technique. The bulk of the bound [3H]-PAF eluted with a higher molecular weight species of PCBP, possibly an aggregated form of PCBP. A smaller amount of [3H]-PAF co-eluted with the major monomeric species of PCBP. Formation of the PCBP-PAF complex was calcium dependent and could be inhibited by phosphorylcholine, suggesting the involvement of the phosphorylcholine binding site on PCBP. Binding of albumin and alpha 1-acid glycoprotein to PAF was not affected by phosphorylcholine or calcium. The specificity of this binding may explain the inhibitory effect of PCBP and related phosphorylcholine binding proteins on PAF induced aggregation of platelets. PMID:2322233

Randell, E; Mookerjea, S; Nagpurkar, A

1990-03-16

224

Inhibition by human serum of lymphocyte proliferation stimulated by purified protein derivative of tuberculin and bacillus Calmette-Guérin.  

PubMed Central

The addition of normal human serum to murine lymphocyte cultures consistently depressed mitogen-induced transformation, as measured by deoxyribonucleic acid synthesis. Stimulation by the B-cell mitogens purified protein derivative-tuberculin, bacillus Calmette-Guérin, and lipopolysaccharide was consistently affected, but there was no inhibition of T-cells when human serum was added to concanavalin A-stimulated cultures. The inhibitory effects were not due to cytotoxic factors for B-lymphocytes or to specific antibodies in serum directed against the mitogens. Analogous results were found with guinea pig serum. Contact of the lymphocytes with the serum within the first 24 h of culture was necessary for inhibition. PMID:387593

Herman-Brand, R; Sultzer, B M

1979-01-01

225

Serum amyloid A is a retinol binding protein that transports retinol during bacterial infection  

PubMed Central

Retinol plays a vital role in the immune response to infection, yet proteins that mediate retinol transport during infection have not been identified. Serum amyloid A (SAA) proteins are strongly induced in the liver by systemic infection and in the intestine by bacterial colonization, but their exact functions remain unclear. Here we show that mouse and human SAAs are retinol binding proteins. Mouse and human SAAs bound retinol with nanomolar affinity, were associated with retinol in vivo, and limited the bacterial burden in tissues after acute infection. We determined the crystal structure of mouse SAA3 at a resolution of 2 Å, finding that it forms a tetramer with a hydrophobic binding pocket that can accommodate retinol. Our results thus identify SAAs as a family of microbe-inducible retinol binding proteins, reveal a unique protein architecture involved in retinol binding, and suggest how retinol is circulated during infection. DOI: http://dx.doi.org/10.7554/eLife.03206.001 PMID:25073702

Derebe, Mehabaw G; Zlatkov, Clare M; Gattu, Sureka; Ruhn, Kelly A; Vaishnava, Shipra; Diehl, Gretchen E; MacMillan, John B; Williams, Noelle S; Hooper, Lora V

2014-01-01

226

Camptothecin-binding site in human serum albumin and protein transformations induced by drug binding.  

PubMed

Circular dichroism (CD) and Raman spectroscopy were employed in order to locate a camptothecin (CPT)-binding site within human serum albumin (HSA) and to identify protein structural transformations induced by CPT binding. A competitive binding of CPT and 3'-azido-3'-deoxythymidine (a ligand occupying IIIA structural sub-domain of the protein) to HSA does not show any competition and demonstrates that the ligands are located in the different binding sites, whereas a HSA-bound CPT may be replaced by warfarin, occupying IIA structural sub-domain of the protein. Raman and CD spectra of HSA and HSA/CPT complexes show that the CPT-binding does not induce changes of the global protein secondary structure. On the other hand, Raman spectra reveal pronounced CPT-induced local structural modifications of the HSA molecule, involving changes in configuration of the two disulfide bonds and transfer of a single Trp-residue to hydrophilic environment. These data suggest that CPT is bound in the region of interdomain connections within the IIA structural domain of HSA and it induces relative movement of the protein structural domains. PMID:9271208

Fleury, F; Ianoul, A; Berjot, M; Feofanov, A; Alix, A J; Nabiev, I

1997-07-14

227

Validation Processes of Protein Biomarkers in Serum—A Cross Platform Comparison  

PubMed Central

Due to insufficient biomarker validation and poor performances in diagnostic assays, the candidate biomarker verification process has to be improved. Multi-analyte immunoassays are the tool of choice for the identification and detailed validation of protein biomarkers in serum. The process of identification and validation of serum biomarkers, as well as their implementation in diagnostic routine requires an application of independent immunoassay platforms with the possibility of high-throughput. This review will focus on three main multi-analyte immunoassay platforms: planar microarrays, multiplex bead systems and, array-based surface plasmon resonance (SPR) chips. Recent developments of each platform will be discussed for application in clinical proteomics, principles, detection methods, and performance strength. The requirements for specific surface functionalization of assay platforms are continuously increasing. The reasons for this increase is the demand for highly sensitive assays, as well as the reduction of non-specific adsorption from complex samples, and with it high signal-to-noise-ratios. To achieve this, different support materials were adapted to the immobilized biomarker/ligand, allowing a high binding capacity and immobilization efficiency. In the case of immunoassays, the immobilized ligands are proteins, antibodies or peptides, which exhibit a diversity of chemical properties (acidic/alkaline; hydrophobic/hydrophilic; secondary or tertiary structure/linear). Consequently it is more challenging to develop immobilization strategies necessary to ensure a homogenous covered surface and reliable assay in comparison to DNA immobilization. New developments concerning material support for each platform are discussed especially with regard to increase the immobilization efficiency and reducing the non-specific adsorption from complex samples like serum and cell lysates. PMID:23112739

Köhler, Katja; Seitz, Harald

2012-01-01

228

Short-term space flight on nitrogenous compounds, lipoproteins, and serum proteins  

NASA Technical Reports Server (NTRS)

Biochemical variables in blood were measured in venous blood samples from 38 to 72 Space Shuttle astronauts before and immediately after flights of 2 to 11 days. Mean pre- and postflight values were compared using the paired t-test or the Wilcoxon signed-rank test. The largest change in serum enzymes was a 21% increase (P = .0014) in gamma-glutamyl-transpeptidase, which may have been related to stress. The median value of apolipoprotein (apo) A-I decreased from 152 to 127 mg/dL (P < .0001), but the change in apo B (77 to 73 mg/dL) was not statistically significant, and the mean apo A-I/apo B ratio remained well above 1.5. A decrease in dietary fat and cholesterol intake during shuttle missions may have been a cause of the change in apo A-I. Twelve of the 16 nonenzyme serum proteins measured were significantly elevated (P < .05), possibly because of hemoconcentration and increased protein catabolism. The 56% increase in haptoglobin may be related to release of suppressed erythropoiesis at landing.

Leach, C. S.; Lane, H. W.; Krauhs, J. M.

1994-01-01

229

Preparation of protein imprinted materials by hierarchical imprinting techniques and application in selective depletion of albumin from human serum  

NASA Astrophysics Data System (ADS)

Hierarchical imprinting was developed to prepare the protein imprinted materials, as the artificial antibody, for the selective depletion of HSA from the human serum proteome. Porcine serum albumin (PSA) was employed as the dummy template for the fabrication of the recognition sites. To demonstrate the advantages of the hierarchical imprinting, molecularly imprinted polymers prepared by hierarchical imprinting technique (h-MIPs) were compared with those obtained by bulk imprinting (b-MIPs), in terms of the binding capacity, adsorption kinetics, selectivity and synthesis reproducibility. The binding capacity of h-MIPs could reach 12 mg g-1. And saturation binding could be reached in less than 20 min for the h-MIPs. In the protein mixture, h-MIPs exhibit excellent selectivity for PSA, with imprinting factors as about 3.6, much higher than those for non-template proteins. For the proteomic application, the identified protein group number in serum treated by h-MIPs was increased to 422, which is 21% higher than that obtained from the original serum, meanwhile the identified protein group number for the Albumin Removal kit was only 376. The results demonstrate that protein imprinted polymers prepared by hierarchical imprinting technique, might become the artificial antibodies for the selective depletion of high abundance proteins in proteome study.

Liu, Jinxiang; Deng, Qiliang; Tao, Dingyin; Yang, Kaiguang; Zhang, Lihua; Liang, Zhen; Zhang, Yukui

2014-06-01

230

Single chain variable fragment displaying M13 phage library functionalized magnetic microsphere-based protein equalizer for human serum protein analysis.  

PubMed

Single chain variable fragment (scFv) displaying the M13 phage library was covalently immobilized on magnetic microspheres and used as a protein equalizer for the treatment of human serum. First, scFv displaying M13 phage library functionalized magnetic microspheres (scFv@M13@MM) was incubated with a human serum sample. Second, captured proteins on scFv@M13@MM were eluted with 2 M NaCl, 50 mM glycine-hydrochloric acid (Gly-HCl), and 20% (v/v) acetonitrile with 0.5% (v/v) trifluoroacetic acid in sequence. Finally, the tightly bonded proteins were released by the treatment with thrombin. The eluates were first analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with silver staining. Results indicated that the difference of protein concentration was reduced obviously in NaCl and Gly-HCl fractions compared with untreated human serum sample. The eluates were also digested with trypsin, followed by online 2D-strong cation exchange (SCX)-RPLC-ESI-MS/MS analysis. Results demonstrated that the number of proteins identified from an scFv@M13@MM treated human serum sample was improved 100% compared with that from the untreated sample. In addition, the spectral count of 10 high abundance proteins (serum albumin, serotransferrin, ?-2-macroglobulin, ?-1-antitrypsin, apolipoprotein B-100, Ig ?-2 chain C region, haptoglobin, hemopexin, ?-1-acid glycoprotein 1, and ?-2-HS-glycoprotein) decreased evidently after scFv@M13@MM treatment. All these results demonstrate that scFv@M13@MM could efficiently remove high-abundance proteins, reduce the protein concentration difference of human serum, and result in more protein identification. PMID:22909037

Zhu, Guijie; Zhao, Peng; Deng, Nan; Tao, Dingyin; Sun, Liangliang; Liang, Zhen; Zhang, Lihua; Zhang, Yukui

2012-09-18

231

Differential Denaturation of Serum Proteome Reveals a Significant Amount of Hidden Information in Complex Mixtures of Proteins  

PubMed Central

Recently developed proteomic technologies allow to profile thousands of proteins within a high-throughput approach towards biomarker discovery, although results are not as satisfactory as expected. In the present study we demonstrate that serum proteome denaturation is a key underestimated feature; in fact, a new differential denaturation protocol better discriminates serum proteins according to their electrophoretic mobility as compared to single-denaturation protocols. Sixty nine different denaturation treatments were tested and the 3 most discriminating ones were selected (TRIDENT analysis) and applied to human sera, showing a significant improvement of serum protein discrimination as confirmed by MALDI-TOF/MS and LC-MS/MS identification, depending on the type of denaturation applied. Thereafter sera from mice and patients carrying cutaneous melanoma were analyzed through TRIDENT. Nine and 8 protein bands were found differentially expressed in mice and human melanoma sera, compared to healthy controls (p<0.05); three of them were found, for the first time, significantly modulated: ?2macroglobulin (down-regulated in melanoma, p<0.001), Apolipoprotein-E and Apolipoprotein-A1 (both up-regulated in melanoma, p<0.04), both in mice and humans. The modulation was confirmed by immunological methods. Other less abundant proteins (e.g. gelsolin) were found significantly modulated (p<0.05). Conclusions: i) serum proteome contains a large amount of information, still neglected, related to proteins folding; ii) a careful serum denaturation may significantly improve analytical procedures involving complex protein mixtures; iii) serum differential denaturation protocol highlights interesting proteomic differences between cancer and healthy sera. PMID:23533572

Polci, Maria Letizia; Rossi, Stefania; Cordella, Martina; Carlucci, Giuseppe; Marchetti, Paolo; Antonini-Cappellini, Giancarlo; Facchiano, Antonio; D'Arcangelo, Daniela; Facchiano, Francesco

2013-01-01

232

Development of an ELISA detecting Tumor Protein 53-Induced Nuclear Protein 1 in serum of prostate cancer patients.  

PubMed

Tumor Protein 53-Induced Nuclear Protein 1 (TP53INP1) plays an important role during cell stress response in synergy with the potent "genome-keeper" p53. In human, the gene encoding TP53INP1 is expressed at very high level in some pathological situations, such as inflammation and prostate cancer (PC). TP53INP1 overexpression in PC seems to be a worse prognostic factor, particularly predictive of biological cancer relapse, making TP53INP1 a relevant specific target for molecular therapy of Castration Resistant (CR) PC. In that context, detection of TP53INP1 in patient biological fluids is a promising diagnostic avenue. We report here successful development of a new Enzyme-Linked Immunosorbent Assay (ELISA) detecting TP53INP1, taking advantage of molecular tools (monoclonal antibodies (mAbs) and recombinant proteins) generated in the laboratory during the course of basic functional investigations devoted to TP53INP1. The ELISA principle is based on a sandwich immunoenzymatic system, TP53INP1 protein being trapped by a first specific mAb coated on microplate then recognized by a second specific mAb. This new assay allows specific detection of TP53INP1 in serum of several PC patients. This breakthrough paves the way towards investigation of a large cohort of patients and assessment of clinical applications of TP53INP1 dosage. PMID:24600558

Saadi, Houda; Seillier, Marion; Sandi, Maria José; Peuget, Sylvain; Kellenberger, Christine; Gravis, Gwenaëlle; Dusetti, Nelson J; Iovanna, Juan L; Rocchi, Palma; Amri, Mohamed; Carrier, Alice

2013-01-01

233

Increased serum levels of macrophage inflammatory protein-3? and cystatin a predict a poor prognosis of nasopharyngeal carcinoma.  

PubMed

This study was aimed to investigate the roles of serum macrophage inflammatory protein-3? (MIP-3?) and cystatin A in nasopharyngeal carcinoma (NPC) prognosis.The serum levels of MIP-3? and cystatin A in 140 primary NPC patients without distant metastasis were detected by enzyme-linked immunosorbent assay before and after treatment. The results were compared with those in 100 healthy controls. The log-rank test was used to compare survival curves of the 2 groups. Multivariate analysis of prognostic factors used Cox proportional hazards regression model.Serum levels of MIP-3? and cystatin A in pretreatment patients with NPC were higher than those in healthy controls. Concentrations of these 2 factors in the majority of patients after the therapy decreased to control level. Patients with high serum level of MIP-3? and cystatin A before treatment had poorer overall survival (OS), local recurrence-free survival, and distant metastasis-free survival than the ones with low level. In addition, serum pretreatment MIP-3? and cystatin A levels were independent prognostic factors for OS and distant metastasis-free survival of NPC patients; serum posttreatment MIP-3? and cystatin A levels were independent prognostic factors of local recurrence-free survival.Our results revealed that serum MIP-3? and cystatin A may be promising candidate prognostic factors for NPC, and higher serum levels of MIP-3? and cystatin A correlate with shorter probability of OS, local recurrence, and distant metastasis. PMID:25396333

Cai, Yonglin; Li, Jun; Lu, Aiying; Zhong, Weiming; Gao, Jianquan; Zheng, Yuming; Zeng, Hong; Wang, Wei; Tang, Minzhong

2014-11-01

234

Tuberous Sclerosis Complex Proteins 1 and 2 Control Serum-Dependent Translation in a TOP-Dependent and Independent Manner  

Microsoft Academic Search

The tuberous sclerosis complex (TSC) proteins TSC1 and TSC2 regulate protein translation by inhibiting the serine\\/threonine kinase mTORC1 (for mammalian target of rapamycin complex 1). However, how TSC1 and TSC2 control overall protein synthesis and the translation of specific mRNAs in response to different mitogenic and nutritional stimuli is largely unknown. We show here that serum withdrawal inhibits mTORC1 signaling,

Benoit Bilanges; Rhoda Argonza-Barrett; Marina Kolesnichenko; Christina Skinner; Manoj Nair; Michelle Chen; David Stokoe

2007-01-01

235

Serum Protein and Casein Concentration: Effect on pH and Freezing Point of Milk with Added CO21  

Microsoft Academic Search

The objective of this studywas to determine the effect of protein concentration and protein type (i.e., casein (CN) and serum protein (SP)) on pH (0°C) and freezing point (FP) of skim milk upon CO2 injection at 0°C. CN- free skim milks with increasing SP content (0, 3, and 6%) and skim milks with the same SP content (0.6%) but increasing

Y. Ma; D. M. Barbano

2003-01-01

236

A time-resolved immunoassay to measure serum antibodies to the rotavirus VP6 capsid protein  

PubMed Central

The rotavirus (RV) inner capsid protein VP6 is widely used to evaluate immune response during natural infection and in vaccine studies. Recombinant VP6 from the most prevalent circulating rotavirus strains in each subgroup (SG) identified in a birth cohort of children in southern India [SGII (G1P[8]) and SGI (G10P[11])] were produced. The purified proteins were used to measure VP6-specific antibodies in a Dissociation-Enhanced Lanthanide Fluorometric Immunoassay (DELFIA). The ability of the assay to detect a ?2 fold rise in IgG level in a panel of serum samples from a longitudinal study was compared to a gold standard virus-capture ELISA. A strong association was observed between the assays (p < 0.001; chi-squared test) with assay performances remaining similar when the samples were subdivided as having a fold change increase in VP6 antibody levels (a) within 90 days of RV RNA detection in stool or (b) if no RV RNA was detected within that time period. This study demonstrates the suitability of using recombinant proteins to measure anti-RV immune responses and serves as a “proof of principle” to examine the antibody responses generated to other recombinant RV proteins and thereby possibly identify a correlate of protection. PMID:23183143

Kavanagh, Owen; Zeng, Xi-Lei; Ramani, Sasirekha; Mukhopadhya, Indrani; Crawford, Sue E.; Kang, Gagandeep; Estes, Mary K.

2013-01-01

237

Monte carlo method-based QSAR modeling of penicillins binding to human serum proteins.  

PubMed

The binding of penicillins to human serum proteins was modeled with optimal descriptors based on the Simplified Molecular Input-Line Entry System (SMILES). The concentrations of protein-bound drug for 87 penicillins expressed as percentage of the total plasma concentration were used as experimental data. The Monte Carlo method was used as a computational tool to build up the quantitative structure-activity relationship (QSAR) model for penicillins binding to plasma proteins. One random data split into training, test and validation set was examined. The calculated QSAR model had the following statistical parameters: r(2) ?=?0.8760, q(2) ?=?0.8665, s?=?8.94 for the training set and r(2) ?=?0.9812, q(2) ?=?0.9753, s?=?7.31 for the test set. For the validation set, the statistical parameters were r(2) ?=?0.727 and s?=?12.52, but after removing the three worst outliers, the statistical parameters improved to r(2) ?=?0.921 and s?=?7.18. SMILES-based molecular fragments (structural indicators) responsible for the increase and decrease of penicillins binding to plasma proteins were identified. The possibility of using these results for the computer-aided design of new penicillins with desired binding properties is presented. PMID:25408278

Veselinovi?, Jovana B; Toropov, Andrey A; Toropova, Alla P; Nikoli?, Goran M; Veselinovi?, Aleksandar M

2015-01-01

238

Serum proteins and some biochemical parameters in broiler chickens fed with raw and treated bitter vetch (Vicia ervilia) seeds.  

PubMed

This study carried out to evaluate the effect of bitter vetch seeds on serum proteins and biochemical parameters in broiler chickens. A total of 1320 one-day-old broiler chicks of a commercial breed were placed in 64 pens. Treatments were included raw and four different processed bitter vetch seeds in three levels (150, 300 and 450 g kg(-1)) and a corn-soybean based diet as control. Each treatment group consisted of four replicates. Processing methods were included soaked in water for 12 h, autoclaved, then dried at room temperature (SAD); ground, soaked in water for 24 h, autoclaved and dried (GSAD); ground, soaked in water for 47 h with exchange water every 12 h, cooked and dried (GSCD) and ground, soaked at 1% acetic acid solution for 24 h at 60 degrees C (AA). Feeding raw, AA and GSAD seeds decreased serum albumin significantly (p<0.05) in 21-days-old chicks. Chickens that fed with raw and treated bitter vetch seed had lower alpha 1 and gamma globulins than control (p<0.05). Increasing raw and treated bitter vetch seeds from 15 to 30 and 45% decreased albumin, alpha 1 and gamma globulins and increased alpha 2 and beta globulins significantly (p<0.05). In 14-days-old chicks feeding raw and treated biter vetch had no effect on serum urea, but uric acid concentration decreased significantly (p<0.05). Feeding SAD seeds increased serum urea significantly (p<0.05), but uric acid concentration did not change with feeding raw and treated bitter vetch seeds in 42-day-old chicks. Adding raw and treated bitter vetch seeds to diet increased T4 and decreased T3 concentrations in all ages. At 28-days-old chicks, feeding raw and treated biter vetch seeds decreased alkaline phosphatase concentration significantly than control. Results showed that raw bitter vetch seeds have some toxic effects on metabolism in broiler chickens and GSCD and SAD treatments were more effective to detoxification of this seed. PMID:19069902

Sadeghi, Gh; Pourreza, J

2007-03-15

239

An electrochemical immunosensor to minimize the nonspecific adsorption and to improve sensitivity of protein assays in human serum.  

PubMed

An electrochemical immunoassay which minimizes nonspecific protein adsorption and improves detection sensitivity of proteomic cancer biomarker is described. Our technique comprises two novel features: (i) a high density terminally functionalized poly(N-isopropyl acrylamide) 'brush' layer is grown by surface initiated reversible addition fragmentation chain transfer (RAFT) polymerization method from the electrode surface in order to minimize nonspecific adsorption of serum proteins and other biomolecules, and (ii) a signal amplifying 'bionanoconjugate' comprised of graphene oxide nanosheets decorated with CdSe quantum dots and recombinant single-chain variable fragments towards MSLN, is used to 'physically' amplify the anodic stripping voltammetric signal. This method enabled a detection limit of ca. 1 pg/mL MSLN (RSD=4.6%, n=4) spiked in serum samples. Because of the simple, specific and sensitive nature of this methodology, we feel that it may find potential use in serum-based protein diagnostics. PMID:22705407

Shiddiky, Muhammad J A; Kithva, Prakash H; Kozak, Darby; Trau, Matt

2012-01-01

240

Nitric oxide protects neuroblastoma cells from apoptosis induced by serum deprivation through cAMP-response element-binding protein (CREB) activation.  

PubMed

The transcription factor cAMP-response element-binding protein (CREB) mediates survival in many cells, including neurons. Recently, death of cerebellar granule neurons due to nitric oxide (NO) deprivation was shown to be accompanied by down-regulation of CREB activity (). We now provide evidence that overproduction of endogenous NO or supplementation with exogenous NO renders SK-N-BE human neuroblastoma cells more resistant to apoptosis induced by serum deprivation. Parental cells underwent apoptosis after 24 h of serum deprivation, an outcome largely absent in clones overexpressing human neuronal nitric oxide synthase (nNOS). This protective effect was reversed by the inhibition of NOS itself or soluble guanylyl cyclase, pointing at cGMP as an intermediate effector of NO-mediated rescue. A slow-releasing NO donor protected parental cells to a significant extent, thus confirming the survival effect of NO. The impaired viability of serum-deprived parental cells was accompanied by a strong decrease of CREB phosphorylation and transcriptional activity, effects significantly attenuated in nNOS-overexpressing clones. To confirm the role of CREB in survival, the ectopic expression of CREB and/or protein kinase A largely counteracted serum deprivation-induced cell death of SK-N-BE cells, whereas transfection with a CREB negative mutant was ineffective. These experiments indicate that CREB activity is an important step for NO-mediated survival in neuronal cells. PMID:12368293

Ciani, Elisabetta; Guidi, Sandra; Della Valle, Giuliano; Perini, Giovanni; Bartesaghi, Renata; Contestabile, Antonio

2002-12-20

241

Gender- or age-related binding characteristics of valproic acid to serum proteins in adult patients with epilepsy.  

PubMed

The aim of the present study was to determine the gender- or age-related binding characteristics of valproic acid (VPA) to serum proteins in the adult population. Serum samples examined in the study were obtained from 70 adult patients (36 males, 34 females) with epilepsy on VPA monotherapy. Their age ranged from 16 to 68 years (mean age with (SD), 37.7 (15.7) years; <45 years, n=44; >/=45 years, n=26). The in vivo population binding parameters of VPA to serum proteins and theoretical minimal unbound serum VPA fraction (Fu) were determined using an equation derived from the Scatchard equation in: (1), all; (2), male and female subgroups; and (3), younger (<45 years) and older (>/=45 years) subgroups. There was a significant difference in serum concentration of unbound VPA between male and female patients. The mean association constant (K) was 0.010 microM(-1) in all, male, and female patients. The mean total concentration of binding sites (n(Pt)) was 1453 microM for all patients, and 1561 and 1394 microM for male and female patients, respectively. The Fu was 0.064 for all patients, and 0.060 and 0.067 for male and female patients, respectively. There were no significant differences in the binding characteristics of VPA to serum proteins between the male and female groups. On the other hand, there were significant differences in the serum albumin concentration and molar concentration ratio of free fatty acids to albumin in serum between the younger and older patients. The mean value of K was 0.016 microM(-1) for the younger patients and 0.007 microM (-1) for the older patients. The mean n(Pt) was 1157 microM for the younger patients and 1703 microM for the older patients. The Fu was 0.051 for the younger patients and 0.077 for the older patients. Thus, significant differences were observed in the binding characteristics of VPA to serum proteins between the younger and older groups. Our results show that age, but not gender, has significant influences on the binding characteristics of VPA to serum proteins in our patient population. PMID:11438424

Kodama, Y; Kodama, H; Kuranari, M; Tsutsumi, K; Ono, S; Yukawa, E; Fujimura, A

2001-07-01

242

Interaction of rat serum phosphorylcholine-binding protein with phospholipid-containing liposomes.  

PubMed

Rat serum phosphorylcholine-binding protein (PCBP) has been shown to inhibit the Ca2+-modulated heparin-lipoprotein precipitation reaction. This effect of PCBP on the reaction is prevented by phosphorylcholine (P-choline). A stoichiometric relationship between the serum very low density lipoproteins and PCBP was also evident in the heparin-very low density lipoprotein precipitation reaction (Nagpurkar, A., and Mookerjea, S. (1981) J. Biol. Chem. 256, 7440-7448). A study on the binding of PCBP to artificial liposomes was initiated to understand the mechanism of interaction between PCBP and phospholipids in soluble lipoproteins. Radioiodinated PCBP was incubated with multilamellar liposomes prepared with egg phosphatidylcholine (PC) and lysophosphatidylcholine, and it was found that the binding of PCBP to multilamellar liposomes was Ca2+-dependent and required the incorporation of about 25% lysophosphatidylcholine into the liposomes. Furthermore, the binding could be inhibited by addition of P-choline. The optimum concentrations of Ca2+ and liposomes as well as time and temperature required for binding were established. Analysis of Scatchard binding data yielded an association constant K alpha of 1.8 X 10(6) M-1 and a total binding capacity of 0.96 nmol/mumol of phospholipid. Substitution of P-choline head groups by phosphorylethanolamine and phosphorylserine on the PC of liposomes reduced the binding considerably, whereas the substitution of fatty acyl moieties on the PC of liposomes was without any effect. Bovine serum albumin was required in the assay to prevent artifactual binding of 125I-PCBP to the assay tubes. The results suggest that the P-choline groups on the surface of liposomes play an important role in the binding to PCBP and this may provide a possible explanation of the effect of PCBP on the Ca2+-dependent heparin-lipoprotein precipitation reaction. PMID:6885790

Nagpurkar, A; Saxena, U; Mookerjea, S

1983-09-10

243

Clinical value of serum eosinophilic cationic protein assessment in children with inflammatory bowel disease  

PubMed Central

Introduction Eosinophils contribute to the pathogenesis of inflammatory bowel disease (IBD) in the intestine. Eosinophilic cationic protein (ECP) is one of the most important eosinophilic specific mediators released during activation. The aim of the study was to evaluate the clinical value of serum ECP determination in children with active and inactive IBD and its correlation with disease activity. Material and methods There were 125 children with IBD (63 with Crohn's disease – CD, 44 with ulcerative colitis – UC, 18 indeterminate colitis – IC) enrolled in the study. Among them 83 children were in the active phase of the disease, while the remaining 42 were in remission. The control group consisted of 56 healthy children. The ECP was assessed three times in children with active IBD, at baseline and after 2 and 6 weeks of treatment and once in children with inactive IBD and controls using fluoroenzymeimmunoassays. Results We found elevated ECP at baseline in the total active IBD group when compared to the inactive IBD and control groups, decreasing during treatment. Serum ECP was also elevated in the active UC and CD groups when compared to the inactive UC and CD groups, and correlated with clinical UC and CD activity (R = 0.57 and R = 0.52, p < 0.05, respectively) and duration of the clinical manifestation in UC (R = 0.62, p < 0.05) but not with the disease location in the gastrointestinal tract, or endoscopic and histopathological activity. Conclusions Evaluation of serum ECP in children with IBD may be useful in disease activity assessment at onset and during the treatment.

Tomasik, Przemys?aw; Pieczarkowski, Stanis?aw; Kowalska-Duplaga, Kinga; Grzenda-Adamek, Zofia; Fyderek, Krzysztof

2013-01-01

244

Expression cloning of a novel zinc finger protein that binds to the c-fos serum response element.  

PubMed Central

Induction of c-fos transcription by serum growth factors requires the serum response element (SRE). The SRE is a multifunctional element which responds to several positively and negatively acting signals. To identify cellular proteins that might mediate functions of the SRE, we screened a human cDNA expression library with an SRE probe. We report the isolation and characterization of SRE-ZBP, a previously unidentified SRE-binding protein. SRE-ZBP is a member of the C2H2 zinc finger family of proteins exemplified by TFIIIA and the Drosophila Krüppel protein. The seven tandemly repeated zinc finger motifs in SRE-ZBP are sufficient for high-affinity binding to the SRE. We show that SRE-ZBP is a nuclear protein and identify a candidate cellular protein encoded by the SRE-ZBP gene. Because we cannot detect any DNA-binding activity attributable to the endogenous protein, we propose that SRE-ZBP activity may be subject to posttranslational regulation. Like c-fos mRNA, SRE-ZBP mRNA is serum inducible in HeLa cells, but with slower kinetics. The role of SRE-ZBP in the regulation of c-fos transcription remains unestablished, but this protein binds to a region of the SRE where mutations lead to derepression. Images PMID:1569959

Attar, R M; Gilman, M Z

1992-01-01

245

Muscle enzyme and fiber type-specific sarcomere protein increases in serum after inertial concentric-eccentric exercise.  

PubMed

Muscle damage induced by inertial exercise performed on a flywheel device was assessed through the serum evolution of muscle enzymes, interleukin 6, and fiber type-specific sarcomere proteins such as fast myosin (FM) and slow myosin (SM). We hypothesized that a model of muscle damage could be constructed by measuring the evolution of serum concentration of muscle proteins following inertial exercise, according to their molecular weight and the fiber compartment in which they are located. Moreover, by measuring FM and SM, the type of fibers that are affected could be assessed. Serum profiles were registered before and 24, 48, and 144?h after exercise in 10 healthy and recreationally active young men. Creatine kinase (CK) and CK-myocardial band isoenzyme increased in serum early (24?h) and returned to baseline values after 48?h. FM increased in serum late (48?h) and remained elevated 144?h post-exercise. The increase in serum muscle enzymes suggests increased membrane permeability of both fast and slow fibers, and the increase in FM reveals sarcomere disruption as well as increased membrane permeability of fast fibers. Consequently, FM could be adopted as a fiber type-specific biomarker of muscle damage. PMID:25441613

Carmona, G; Guerrero, M; Cussó, R; Padullés, J M; Moras, G; Lloret, M; Bedini, J L; Cadefau, J A

2014-12-01

246

A Closer Look at Evolution: Variants (SNPs) of Genes Involved in Skin Pigmentation, Including EXOC2, TYR, TYRP1, and DCT, Are Associated With 25(OH)D Serum Concentration.  

PubMed

Vitamin D deficiency is common in the Caucasian population and is associated with increased incidence and unfavorable outcome of many diseases, including various types of cancer, infectious, cardiovascular, and autoimmune diseases. Individual factors that predispose for a person's vitamin D status, such as skin type, have been identified, but limited data exist on genetic determinants of serum 25-hydroxyvitamin D (25[OH]D) concentration. We have tested the hypothesis that variants of genes (single nucleotide polymorphisms [SNPs]) involved in skin pigmentation are predictive of serum 25(OH)D levels. Serum 25(OH)D and SNPs (n = 960) related to genes with relevance for skin pigmentation (tyrosinase [TYR], TYR-related protein 1 [TYRP1], dopachrome tautomerase [DCT], oculocutaneous albinism II [OCA2], two pore segment channel 2 [TPCN2], solute carrier family 24 A4 [SLC24A4], solute carrier family 45 A2 [SLC45A2], agouti signalling peptide [ASIP], cyclic AMP-dependent transcription factor [ATF1], microphthalmia-associated transcription factor [MITF], proopiomelanocortin [POMC], cAMP-dependent protein kinase catalytic subunit beta [PRKACB], cAMP-dependent protein kinase catalytic subunit gamma [PRKACG], cAMP-dependent protein kinase type I-alpha regulatory subunit [PRKAR1A], cAMP-dependent protein kinase type II-alpha regulatory subunit [PRKAR2A], cAMP-dependent protein kinase type II-beta regulatory subunit [PRKAR2B], tubulin beta-3 chain/melanocortin receptor 1 [TUBB3/MC1R], Cadherin-1 [CDH1], catenin beta 1 [CTNNB1], Endothelin 1 [EDN1], endothelin 3 [EDN3], endothelin receptor type B [EDNRB], fibroblast growth factor 2 [FGF2], KIT, KIT ligand [KITLG], nerve growth factor [NGF], interferon regulatory factor 4 [IRF4], exocyst complex component 2 [EXOC2], and tumor protein 53 [TP53]) were analyzed in a cohort of participants of the Ludwigshafen Risk and Cardiovascular Health Study (n = 2970). A total of 46 SNPs were associated (P <.05) with lower or higher serum 25(OH)D levels as compared with the total cohort (median, 15.5 ng/mL). Although 1 SNP in the EXOC2 gene reached the aimed significance level after correction for multiple comparisons (false discovery rate) and was associated with a ?25(OH)D value more than 5.00 ng/mL, 11 SNPs located in the TYR (n = 4), PRKACG (n = 1), EDN1 (n = 3), TYRP1 (n = 1), and microphthalmia-associated transcription factor (n = 2) genes reached the aimed significance level after false discovery rate correction but were not associated with ?25(OH)D value more than 5.00 ng/mL. We conclude that variants of genes involved in skin pigmentation are predictive of serum 25(OH)D levels in the Caucasian population. Our data indicate that out of the variants in 29 different genes analyzed, variants of 11 genes, including EXOC2, TYR, and TYRP1, have the highest impact on vitamin D status. Our results have a fundamental importance to understand the role of sunlight, skin pigmentation, and vitamin D for the human evolution. PMID:25396269

Saternus, Roman; Pilz, Stefan; Gräber, Stefan; Kleber, Marcus; März, Winfried; Vogt, Thomas; Reichrath, Jörg

2015-01-01

247

Decreased serum protein associated with Mycobacterium avium subspecies paratuberculosis shedding in German Holstein cows.  

PubMed

Using well established metabolic parameters, this study aimed to substantiate differences in protein and energy metabolism between Mycobacterium avium subspecies paratuberculosis (MAP) positive and negative dairy cows tested by faecal culture. A total of 227 MAP-positive and 239 MAP-negative German Holstein cows kept in 13 MAP-positive dairy herds were selected for metabolic testing. The serum concentrations of total protein (TP), bilirubin, cholesterol and betahydroxybutyrate were measured as well as the activities of Glutamate-Dehydrogenase (GLDH) and Aspartate-Aminotransferase. MAP-positive cows were characterised by a decreased mean TP (66.5 g/l) compared to the MAP-negative controls (73.2 g/l). Mean log10 GLDH activities tended to be higher in MAP-positive than MAP-negative cows. Concerning TP, there was a significant interaction between MAP status and farm. Within four farms, the difference between MAP-positive and MAP-negative animals differed significantly, while in the other farms this difference was not significant. It is concluded that a decreased TP and an increased GLDH indicate alterations in protein metabolism. These findings suggest an enhanced liver cell turnover in MAP-positive cows. The results contribute to an understanding of the metabolic alterations in MAP-positive dairy cows. PMID:24578317

Donat, K; Erhardt, G; Soschinka, A; Brandt, H R

2014-04-19

248

Protective effect of serum antibodies against a 110-kilodalton protein of Actinobacillus actinomycetemcomitans following periodontal therapy.  

PubMed

Thirty-four adult patients with untreated periodontitis were randomly assigned to receive full mouth scaling alone or scaling with an adjunctive antimicrobial therapy, both followed by supportive periodontal therapy. At 24 months, specific serum immunoglobulin A (IgA), IgG and IgG subclass antibody reactivities against a 110-kDa protein of Actinobacillus actinomycetemcomitans were assessed by Western blot. In patients harboring A. actinomycetemcomitans intraorally, the IgG4 antibody reactivity against the 110-kDa protein of A. actinomycetemcomitans was associated with significantly increased survival rates of teeth and of sites not exhibiting 2 mm or more of probing attachment loss. The same trend was found for IgG3 and IgG2 antibody reactivities, but it was statistically insignificant. No association with clinical treatment outcome was observed for IgA, IgG and IgG1 antibody reactivities. The results indicated that systemic IgG4 antibody reactivity against the 110-kDa protein of A. actinomycetemcomitans may have a protective effect against periodontal disease progression in patients harboring A. actinomycetemcomitans and receiving periodontal therapy. PMID:10551154

Beikler, T; Karch, H; Ehmke, B; Klaiber, B; Flemmig, T F

1999-10-01

249

Serum protein profiling of adults and children with Crohn’s disease  

PubMed Central

Objectives Crohn’s disease (CD) and ulcerative colitis (UC), known collectively as inflammatory bowel disease (IBD), are chronic immuno-inflammatory pathologies of unknown etiology. Despite the frequent utilization of biomarkers in medical practice, there is a relative lack of information regarding validated paediatric biomarkers for IBD. Further, biomarkers proved to be efficacious in adults are frequently extrapolated to the paediatric clinical setting without considering that the pathogenesis of many diseases is distinctly different in children. In the current study, proteomics technology was employed in order to monitor differences in protein expression among adult and children CD patients, in order to identify a panel of candidate protein biomarkers that might be used to improve prognostic-diagnostic accuracy and to advance paediatric medical care. Methods Male and female serum samples from 12 adults and 12 children with active CD were subjected to two-dimensional gel electrophoresis. Following the relative quantitation of protein spots exhibiting a differential expression between the two groups by densitometry, the spots were further characterized by MALDI-TOF-MS. Results were confirmed by Western blot analysis. Results Clusterin (CLUS) was found to be significantly over-expressed in adults with CD, whereas ceruloplasmin (CERU) and apolipoprotein B-100 (APOB) were found to be significantly over-expressed in children indicating that the expression of these proteins might be implicated in the onset or progression of CD in these two sub-groups of patients. Conclusions Interestingly, we found a differential expression of several proteins in adults versus paediatric CD patients. Undoubtedly, future experiments using a larger cohort of CD patients are needed to evaluate the relevance of our preliminary findings. PMID:25250685

Vaiopoulou, Anna; Gazouli, Maria; Papadopoulou, Aggeliki; Anagnostopoulos, Athanassios K; Karamanolis, George; Theodoropoulos, George E.; M’Koma, Amosy; Tsangaris, George T.

2014-01-01

250

Including food 25-hydroxyvitamin D in intake estimates may reduce the discrepancy between dietary and serum measures of vitamin D status.  

PubMed

The discrepancy between the commonly used vitamin D status measures-intake and serum 25-hydroxyvitamin D [25(OH)D] concentrations--has been perplexing. Sun exposure increases serum 25(OH)D concentrations and is often used as an explanation for the higher population-based serum concentrations in the face of apparently low vitamin D intake. However, sun exposure may not be the total explanation. 25(OH)D, a metabolite of vitamin D, is known to be present in animal-based foods. It has been measured and reported only sporadically and is not currently factored into U.S. estimates of vitamin D intake. Previously unavailable preliminary USDA data specifying the 25(OH)D content of a subset of foods allowed exploration of the potential change in the reported overall vitamin D content of foods when the presence of 25(OH)D was included. The issue of 25(OH)D potency was addressed, and available commodity intake estimates were used to outline trends in projected vitamin D intake when 25(OH)D in foods was taken into account. Given the data available, there were notable increases in the total vitamin D content of a number of animal-based foods when potency-adjusted 25(OH)D was included, and in turn there was a potentially meaningful increase (1.7-2.9 ?g or 15-30% of average requirement) in vitamin D intake estimates. The apparent increase could reduce discrepancies between intake estimates and serum 25(OH)D concentrations. The relevance to dietary interventions is discussed, and the need for continued exploration regarding 25(OH)D measurement is highlighted. PMID:24623845

Taylor, Christine L; Patterson, Kristine Y; Roseland, Janet M; Wise, Stephen A; Merkel, Joyce M; Pehrsson, Pamela R; Yetley, Elizabeth A

2014-05-01

251

L-carnitine infusions may suppress serum C-reactive protein and improve nutritional status in maintenance hemodialysis patients.  

PubMed

Scattered reports indicate that L-carnitine may suppress proinflammatory cytokines in sick individuals without renal disease and may improve protein synthesis or nitrogen balance either in patients without renal disease or in maintenance hemodialysis (MHD) or chronic peritoneal dialysis patients. We conducted an experimental study in MHD patients to evaluate the effects of L-carnitine treatment on inflammatory and protein-energy nutritional status. MHD patients were assigned to receive intravenous injections of L-carnitine 20 mg/kg (n = 48) or placebo (n = 65) thrice weekly at the end of each hemodialysis treatment for 6 months. The carnitine-treated group showed a statistically significant decrease in serum C-reactive protein and increase in serum albumin and transferrin, blood hemoglobin, and body mass index. Conversely, in the placebo-treated group, a significant decrease was reported for serum albumin, serum transferrin, and body mass index, whereas the other considered measures did not change significantly. These preliminary findings suggest that in MHD patients, L-carnitine therapy may suppress inflammation, particularly among those patients with C-reactive protein > or =3 mg/dL, and may improve protein-energy nutritional status. PMID:15827896

Savica, Vincenzo; Santoro, Domenico; Mazzaglia, Giampiero; Ciolino, Franco; Monardo, Paolo; Calvani, Menotti; Bellinghieri, Guido; Kopple, Joel D

2005-04-01

252

Phorbol ester, serum, and rous sarcoma virus transforming gene product induce similar phosphorylations of ribosomal protein S6.  

PubMed Central

The addition of phorbol 12-myristate 13-acetate (PMA), a potent tumor promoter, to serum-starved quiescent chicken embryo fibroblasts (CEF) or C127 murine cells resulted in increased phosphorylation of 40S ribosomal protein S6. The effect of PMA on S6 phosphorylation in quiescent CEF was half-maximal at approximately equal to 100 nM and was readily observed at 16 nM. In addition, S6 phosphorylation was increased in serum-starved CEF incubated with the diacylglycerol derivative, 1-oleoyl-2-acetylglycerol. S6 phosphorylation in PMA-stimulated, serum-stimulated, and serum-starved Rous sarcoma virus-transformed CEF was analyzed by phospho amino acid analysis, two-dimensional polyacrylamide gel electrophoresis, limited proteolysis with V8 protease, and two-dimensional thin-layer electrophoresis of chymotryptic digests. Comparison of S6 phosphorylation by these methods suggests that phosphorylation of S6 stimulated by PMA, serum, or oncogenic transformation with Rous sarcoma virus occurs through common pathways. This is further supported by the observation that the simultaneous addition of PMA and serum to CEF or of either PMA or serum to Rous sarcoma virus-transformed CEF did not significantly further increase the incorporation of phosphate into S6. Images PMID:6093101

Blenis, J; Spivack, J G; Erikson, R L

1984-01-01

253

Investigation of the binding of Cr(III) complexes to bovine and human serum proteins: a proteomic approach.  

PubMed

Chromium (Cr) compounds are widely used in alloys manufacturing and forming processes. One of the main concerns in the use of cobalt-chromium (Co-Cr) alloy-based implants is the long-term fate of Co and Cr ions in the blood, organs, and urine of patients. Our previous studies have shown that Cr(III) forms complexes in different cell culture media, whereas Cr(VI) does not form any detectable structure under the same conditions. Because Cr(VI) is known to be more toxic than Cr(III), we hypothesized that the presence of serum proteins in the molecular structure of Cr(III) may be responsible for the difference in toxicity. We investigated the interaction of the Cr(III) complexes with serum proteins and their internalization by U937 macrophage-like cells. By using a proteomic approach, we showed that in the presence of fetal bovine serum, Cr(III) complexes interacted only with albumin, whereas they interacted mainly with albumin, transferrin, and immunoglobulins (Ig) in the presence of human serum (HS). Cr(III) complexes were more easily engulfed by U937 macrophage-like cells when they were formed with HS. To the best of our knowledge, this is the first report on the formation of Cr(III) complexes in the presence of serum proteins and the interaction of these complexes with U937 macrophage-like cells. (c) 2010 Wiley Periodicals, Inc. J Biomed Mater Res, 2010. PMID:20166223

Tkaczyk, Cathy; Huk, Olga L; Mwale, Fackson; Antoniou, John; Zukor, David J; Petit, Alain; Tabrizian, Maryam

2010-07-01

254

Serum sickness  

MedlinePLUS

... after the first exposure to a medication. Injected proteins such as antithymocyte globulin (used to treat organ transplant rejection) and rituximab (used to treat immune disorders and cancers) can cause serum sickness reactions. Blood products may also cause ...

255

Grafting of bovine serum albumin proteins on plasma-modified polymers for potential application in tissue engineering  

NASA Astrophysics Data System (ADS)

In this work, an influence of bovine serum albumin proteins grafting on the surface properties of plasma-treated polyethylene and poly- l-lactic acid was studied. The interaction of the vascular smooth muscle cells with the modified polymer surface was determined. The surface properties were characterized by X-ray photoelectron spectroscopy, atomic force microscopy, nano-LC-ESI-Q-TOF mass spectrometry, electrokinetic analysis, and goniometry. One of the motivations for this work is the idea that by the interaction of the cell with substrate surface, the proteins will form an interlayer between the cell and the substrate. It was proven that when interacting with the plasma-treated high-density polyethylene and poly- l-lactic acid, the bovine serum albumin protein is grafted on the polymer surface. Since the proteins are bonded to the substrate surface, they can stimulate cell adhesion and proliferation.

Kasálková, Nikola Slepi?ková; Slepi?ka, Petr; Kolská, Zde?ka; Hoda?ová, Petra; Ku?ková, Št?pánka; Švor?ík, Václav

2014-04-01

256

Grafting of bovine serum albumin proteins on plasma-modified polymers for potential application in tissue engineering  

PubMed Central

In this work, an influence of bovine serum albumin proteins grafting on the surface properties of plasma-treated polyethylene and poly-l-lactic acid was studied. The interaction of the vascular smooth muscle cells with the modified polymer surface was determined. The surface properties were characterized by X-ray photoelectron spectroscopy, atomic force microscopy, nano-LC-ESI-Q-TOF mass spectrometry, electrokinetic analysis, and goniometry. One of the motivations for this work is the idea that by the interaction of the cell with substrate surface, the proteins will form an interlayer between the cell and the substrate. It was proven that when interacting with the plasma-treated high-density polyethylene and poly-l-lactic acid, the bovine serum albumin protein is grafted on the polymer surface. Since the proteins are bonded to the substrate surface, they can stimulate cell adhesion and proliferation. PMID:24708858

2014-01-01

257

Examination of the relationship between pathological changes, immunological response and serum protein concentrations in pregnant sheep inoculated with Aspergillus fumigatus  

Microsoft Academic Search

Pregnant sheep inoculated withAspergillus fumigatus conidia developed precipitating and latex agglutinating antibodies to mycelial antigens. The titres of these tended to be higher in those animals developing placental or fetal infection than in those which did not. The concentrations of total serum proteins and of albumin,a, ß and? globulins did not show any consistent changes which could be related to

M. J. Corbel; Carol A. Day; D. J. W. Cole

1980-01-01

258

THE INFLUENCE OF SERUM BINDING PROTEINS AND CLEARANCE ON THE COMPARATIVE RECEPTOR BINDING POTENCY OF ENDOCRINE ACTIVE COMPOUNDS  

EPA Science Inventory

THE INFLUENCE OF SERUM BINDING PROTEINS AND CLEARANCE ON THE COMPARATIVE RECEPTOR BINDING POTENCY OF ENDOCRINE ACTIVE COMPOUNDS. JG Teeguarden1 and HA Barton2. 1ENVIRON International, Ruston LA; 2US EPA, ORD, NHEERL, ETD, Pharmacokinetics Branch, RTP, NC. One measure of th...

259

The level of major proteins and minerals in the blood serum of chickens fed diets with pure cellulose.  

PubMed

The aim of the research was to determine the concentration of total protein and its fractions as well as the concentration of selected mineral components in the blood serum of male broiler chickens Cobb 500 fed diets with different cellulose content. Blood samples were collected for examination from the birds' pterygoid canal veins on their 42 day of age. There was no influence of cellulose preparation on the content of total protein and its fractions: albumins, alfa1-, alfa2-, beta-, gamma-globulins, albumin to globulin ratio, inorganic phosphorus, sodium, potassium, chloride and iron concentrations in blood serum. The highest calcium concentration (P < 0.05) was detected in the blood serum of males fed a diet with the highest cellulose content (0.75-0.95%). Simultaneously, a tendency of increased calcium content was observed along with an increased amount of pure cellulose in diets. The lowest magnesium level (P < 0.05) was observed in the blood of birds fed diets with the lowest amount of cellulose (0.25-0.45%). The magnesium level in the blood of birds fed a diet with higher amounts of cellulose did not considerably differ from the control group. Results from the current study suggest that introduction of a limited amount of pure cellulose into the diet of broiler chickens does not affect total protein concentration and protein fractions but can influence the mineral content in the blood serum. PMID:22428310

Bogus?awska-Tryk, Monika; Szymeczko, Roman; Piotrowska, Anna

2012-01-01

260

Relationship between specific gravity, water content, and serum protein extravasation in various types of vasogenic brain edema  

Microsoft Academic Search

Vasogenic brain edema was induced in cats by cold injury (six animals), brain tumors (five animals), and brain abscesses (six animals). Water and electrolyte content, specific gravity, blood volume, and the amount of extravasated serum proteins were determined in small tissue samples taken from gray and white matter at various distances from the lesion. Edema was strictly confined to the

H.-W. Bothe; W. Bodsch; K.-A. Hossmann

1984-01-01

261

Characterizing the Escherichia coli O157:H7 Proteome Including Protein Associations with Higher Order Assemblies  

PubMed Central

Background The recent outbreak of severe infections with Shiga toxin (Stx) producing Escherichia coli (STEC) serotype O104:H4 highlights the need to understand horizontal gene transfer among E. coli strains, identify novel virulence factors and elucidate their pathogenesis. Quantitative shotgun proteomics can contribute to such objectives, allowing insights into the part of the genome translated into proteins and the connectivity of biochemical pathways and higher order assemblies of proteins at the subcellular level. Methodology/Principal Findings We examined protein profiles in cell lysate fractions of STEC strain 86-24 (serotype O157:H7), following growth in cell culture or bacterial isolation from intestines of infected piglets, in the context of functionally and structurally characterized biochemical pathways of E. coli. Protein solubilization in the presence of Triton X-100, EDTA and high salt was followed by size exclusion chromatography into the approximate Mr ranges greater than 280 kDa, 280-80 kDa and 80-10 kDa. Peptide mixtures resulting from these and the insoluble fraction were analyzed by quantitative 2D-LC-nESI-MS/MS. Of the 2521 proteins identified at a 1% false discovery rate, representing 47% of all predicted E. coli O157:H7 gene products, the majority of integral membrane proteins were enriched in the high Mr fraction. Hundreds of proteins were enriched in a Mr range higher than that predicted for a monomer supporting their participation in protein complexes. The insoluble STEC fraction revealed enrichment of aggregation-prone proteins, including many that are part of large structure/function entities such as the ribosome, cytoskeleton and O-antigen biosynthesis cluster. Significance Nearly all E. coli O157:H7 proteins encoded by prophage regions were expressed at low abundance levels or not detected. Comparative quantitative analyses of proteins from distinct cell lysate fractions allowed us to associate uncharacterized proteins with membrane attachment, potential participation in stable protein complexes, and susceptibility to aggregation as part of larger structural assemblies. PMID:22087229

Pieper, Rembert; Zhang, Quanshun; Clark, David J.; Huang, Shih-Ting; Suh, Moo-Jin; Braisted, John C.; Payne, Samuel H.; Fleischmann, Robert D.; Peterson, Scott N.; Tzipori, Saul

2011-01-01

262

Purification of the precursors to vitelline envelope proteins from serum of Sakhalin taimen, Hucho perryi.  

PubMed

High and low molecular weight vitelline envelope-related proteins (hVERP and lVERP) were purified from serum of vitellogenic female Sakhalin taimen (Hucho perryi) by a combination of ion-exchange, hydroxylapatite and gel filtration chromatography. The molecular weight of hVERP was estimated to be 83 kDa by gel filtration, and 48 kDa and 54 kDa in SDS-PAGE under non-reduced and reduced conditions, respectively. The molecular weight of lVERP was 56 kDa by gel filtration, and 42 kDa and 46 kDa on SDS-PAGE (non-reduced and reduced, respectively). Amino acid composition of hVERP was characterized by high content of proline (15.9%) and glutamic acid (13.8%). The lVERP had high contents of glutamic acid (10.8%) and aspartic acid (10.5%). Specific antibodies against hVERP and against lVERP were prepared by immunizing rabbits. The antiserum to hVERP stained bands corresponding to 98 kDa and 48 kDa of vitelline envelope (VE) in SDS-PAGE without reduction, whereas the antiserum to lVERP immunostained 98 kDa and 42 kDa bands. Both specific antibodies recognized the vitelline envelope of vitellogenic oocytes immunocytochemically. Thus, hVERP and lVERP are precursors to vitelline envelope proteins in this species. PMID:9755486

Shimizu, M; Fujita, T; Hara, A

1998-10-15

263

Serum protein oxidation by diesel exhaust particles: effects on oxidative stress and inflammatory response in vitro.  

PubMed

Considerable evidence shows a key role for protein modification in the adverse effects of chemicals; however, the interaction of diesel exhaust particles (DEP) with proteins and the resulting biological activity remains unclear. DEP and carbon black (CB) suspensions with and without bovine serum albumin (BSA) were used to elucidate the biological effects of air pollutants. The DEP and CB samples were then divided into suspensions and supernatants. Two important goals of the interaction of DEP with BSA were as follows: (1) understanding BSA modification by particles and (2) investigating the effects of particles bound with BSA and the corresponding supernatants on cellular oxidative stress and inflammation. We observed significant free amino groups production was caused by DEP. Using liquid chromatography-mass spectrometry (LC-MS), we observed that BSA was significantly oxidised by DEP in the supernatants and that the peptides ETYGDMADCCEK, MPCTEDYLSLILNR and TVMENFVAFVDK, derived BSA-DEP conjugates, were also oxidised. In A549 cells, DEP-BSA suspensions and the corresponding supernatants reduced 8-hydroxy-2'-deoxyguanosine (8-OHdG) production and increased interleukin-6 (IL-6) levels when compared to DEP solutions without BSA. Our findings suggest that oxidatively modified forms of BSA caused by DEP could lead to oxidative stress and the activation of inflammation. PMID:24161433

Chiang, Ling-Ling; Chen, Hao-Cheng; Lee, Chun-Nin; Chuang, Kai-Jen; Chen, Tzu-Tao; Yeh, Chi-Tai; Wang, Liang-Shun; Lee, Wei-Hua; Lin, Lian-Yu; Tseng, Hsiu-Er; Chuang, Hsiao-Chi

2013-11-25

264

Genome-wide protein QTL mapping identifies human plasma kallikrein as a post-translational regulator of serum uPAR levels  

PubMed Central

The soluble cleaved urokinase plasminogen activator receptor (scuPAR) is a circulating protein detected in multiple diseases, including various cancers, cardiovascular disease, and kidney disease, where elevated levels of scuPAR have been associated with worsening prognosis and increased disease aggressiveness. We aimed to identify novel genetic and biomolecular mechanisms regulating scuPAR levels. Elevated serum scuPAR levels were identified in asthma (n=514) and chronic obstructive pulmonary disease (COPD; n=219) cohorts when compared to controls (n=96). In these cohorts, a genome-wide association study of serum scuPAR levels identified a human plasma kallikrein gene (KLKB1) promoter polymorphism (rs4253238) associated with serum scuPAR levels in a control/asthma population (P=1.17×10?7), which was also observed in a COPD population (combined P=5.04×10?12). Using a fluorescent assay, we demonstrated that serum KLKB1 enzymatic activity was driven by rs4253238 and is inverse to scuPAR levels. Biochemical analysis identified that KLKB1 cleaves scuPAR and negates scuPAR's effects on primary human bronchial epithelial cells (HBECs) in vitro. Chymotrypsin was used as a proproteolytic control, while basal HBECs were used as a control to define scuPAR-driven effects. In summary, we reveal a novel post-translational regulatory mechanism for scuPAR using a hypothesis-free approach with implications for multiple human diseases.—Portelli, M. A., Siedlinski, M., Stewart, C. E., Postma, D. S., Nieuwenhuis, M. A., Vonk, J. M., Nurnberg, P., Altmuller, J., Moffatt, M. F., Wardlaw, A. J., Parker, S. G., Connolly, M. J., Koppelman, G. H., Sayers, I. Genome-wide protein QTL mapping identifies human plasma kallikrein as a post-translational regulator of serum uPAR levels. PMID:24249636

Portelli, Michael A.; Siedlinski, Mateusz; Stewart, Ceri E.; Postma, Dirkje S.; Nieuwenhuis, Maartje A.; Vonk, Judith M.; Nurnberg, Peter; Altmuller, Janine; Moffatt, Miriam F.; Wardlaw, Andrew J.; Parker, Stuart G.; Connolly, Martin J.; Koppelman, Gerard H.; Sayers, Ian

2014-01-01

265

No effect of gender or age on binding characteristics of valproic acid to serum proteins in pediatric patients with epilepsy.  

PubMed

The gender- and age-related binding characteristics of valproic acid to serum proteins were determined in the pediatric population. Serum samples examined in the study were obtained from 61 pediatric patients (28 males, 33 females) with epilepsy on valproic acid monotherapy. Their ages ranged from 1 to 15 years (mean age with [SD]: 7.8 [3.9] years; < 10 years, n = 41; > or = 10 years, n = 20). The in vivo population binding parameters of valproic acid to serum proteins and theoretical minimal unbound serum fraction (fu) of valproic acid were determined in (1) all, (2) male and female subgroups, and (3) prepubescent (< 10 years) and pubescent (> or = 10 years) subgroups. The association constant (K) was approximately 1.4 times higher in male (0.018 L/mumol) than in female (0.013 L/mumol) patients, while the total concentration of binding sites (n(Pt)) was 1.2 times greater in female (1235 mumol/L) than in male (997 mumol/L) patients. The fu was 0.053 and 0.059 for male and female patients, respectively. The value of K was approximately 1.6 times higher in the pubescent (0.019 L/mumol) than in the prepubescent (0.012 L/mumol) patients, while the n(Pt) was 1.2 times higher in the prepubescent (1244 mumol/L) than in the pubescent (1057 mumol/L) patients. The fu was 0.063 for the prepubescent and 0.047 for the pubescent patients. No significant differences were observed in binding characteristics of valproic acid to serum proteins between male and female or younger and older patients. However, the differences in valproic acid binding to serum proteins appear to be relatively larger in binding affinity than in binding capacity between the two groups. Because no significant differences were observed in serum concentrations of total and unbound valproic acid, albumin, or free fatty acids between any subgroups (male and female, younger and older), the results suggest that gender or age may not be factors for the determination of the binding characteristics of valproic acid to serum proteins in pediatric patients. PMID:10516942

Kodama, Y; Kodama, H; Kuranari, M; Tsutsumi, K; Ono, S; Fujimura, A

1999-10-01

266

Correlation of Bone Morphogenetic Protein-2 Levels in Serum and Synovial Fluid with Disease Severity of Knee Osteoarthritis  

PubMed Central

Background This study aimed to investigate the bone morphogenetic protein-2 (BMP-2) levels in serum and synovial fluid (SF) of patients with primary knee osteoarthritis (OA) and to exam its correlation with radiographic and symptomatic severity of the disease. Material/Methods A total of 37 knee OA patients and 20 healthy controls were enrolled in this study. Knee OA radiographic grading was performed according to the Kellgren-Lawrence (KL) grading system by evaluating X-ray changes observed in anteroposterior knee radiography. Symptomatic severity of the disease was evaluated according to the Western Ontario McMaster University Osteoarthritis Index (WOMAC) scores. BMP-2 levels in serum and SF were determined using enzyme-linked immunosorbent assay. Results Serum BMP-2 level in patients with knee OA was higher than that in healthy controls. Knee OA patients with KL grade 4 showed significantly elevated BMP-2 levels in the serum and SF compared with those with KL grade 2 and 3. Knee OA patients with KL grade 3 had significant higher SF levels of BMP-2 than those with KL grade 2. BMP-2 levels in the serum and SF of knee OA patients were both positively correlated with KL grades and WOMAC scores. Conclusions BMP2 levels in serum and SF were closely related to the radiographic and symptomatic severity of knee OA and may serve as an alternative biochemical parameter to determine disease severity of primary knee OA. PMID:25644704

Liu, Yan; Hou, Ruizhi; Yin, Ruofeng; Yin, Weitian

2015-01-01

267

Identification of serum sirtuins as novel noninvasive protein markers for frailty  

PubMed Central

Frailty has emerged as a major health issue among older patients. A consensus on definition and diagnosis is yet to be achieved. Various biochemical abnormalities have been reported in frailty. Activation of sirtuins, a conserved family of NAD-dependent proteins, is one of the many mimics of calorie restriction which improves lifespan and health in experimental animals. In this cross-sectional study, we assessed the circulating sirtuin levels in 119 (59.5%) nonfrail and 81 (40.5%) frail individuals, diagnosed by Fried's criteria. Serum SIRT1, SIRT2, and SIRT3 were estimated by surface plasmon resonance (SPR) and Western blot. Serum sirtuins level in mean+SD; SIRT1 (nonfrail –4.67 ± 0.48 ng/?L; frail – 3.72 ± 0.48 ng/?L; P < 0.0001), SIRT2 (nonfrail – 15.18 ± 2.94 ng/?L; frail – 14.19 ± 2.66 ng/?L; P = 0.016), and SIRT3 (nonfrail-7.72 ± 1.84 ng/?L; frail – 6.12 ± 0.97 ng/?L; P < 0.0001) levels were significantly lower among frail patients compared with the nonfrail. In multivariable regression analysis, lower sirtuins level were significantly associated with frailty after adjusting age, gender, diabetes mellitus, hypertension, cognitive status (Mini Mental State Examination scores) and number of comorbidities. For detecting the optimum diagnostic cutoff value a ROC analysis was carried out. The area under curve for SIRT1 was 0.9037 (cutoff – 4.29 ng/?L; sensitivity – 81.48%; specificity – 79.83%) and SIRT3 was 0.7988 (cutoff – 6.61 ng/?L; sensitivity – 70.37%; specificity – 70.59%). This study shows that lower circulating SIRT1 and SIRT3 levels can be distinctive marker of frailty. PMID:25100619

Kumar, Rahul; Mohan, Navinath; Upadhyay, Ashish Datt; Singh, Amrendra Pratap; Sahu, Vishal; Dwivedi, Sadanand; Dey, Aparajit B; Dey, Sharmistha

2014-01-01

268

Identification of serum sirtuins as novel noninvasive protein markers for frailty.  

PubMed

Frailty has emerged as a major health issue among older patients. A consensus on definition and diagnosis is yet to be achieved. Various biochemical abnormalities have been reported in frailty. Activation of sirtuins, a conserved family of NAD-dependent proteins, is one of the many mimics of calorie restriction which improves lifespan and health in experimental animals. In this cross-sectional study, we assessed the circulating sirtuin levels in 119 (59.5%) nonfrail and 81 (40.5%) frail individuals, diagnosed by Fried's criteria. Serum SIRT1, SIRT2, and SIRT3 were estimated by surface plasmon resonance (SPR) and Western blot. Serum sirtuins level in mean+SD; SIRT1 (nonfrail -4.67 ± 0.48 ng/?L; frail - 3.72 ± 0.48 ng/?L; P < 0.0001), SIRT2 (nonfrail - 15.18 ± 2.94 ng/?L; frail - 14.19 ± 2.66 ng/?L; P = 0.016), and SIRT3 (nonfrail-7.72 ± 1.84 ng/?L; frail - 6.12 ± 0.97 ng/?L; P < 0.0001) levels were significantly lower among frail patients compared with the nonfrail. In multivariable regression analysis, lower sirtuins level were significantly associated with frailty after adjusting age, gender, diabetes mellitus, hypertension, cognitive status (Mini Mental State Examination scores) and number of comorbidities. For detecting the optimum diagnostic cutoff value a ROC analysis was carried out. The area under curve for SIRT1 was 0.9037 (cutoff - 4.29 ng/?L; sensitivity - 81.48%; specificity - 79.83%) and SIRT3 was 0.7988 (cutoff - 6.61 ng/?L; sensitivity - 70.37%; specificity - 70.59%). This study shows that lower circulating SIRT1 and SIRT3 levels can be distinctive marker of frailty. PMID:25100619

Kumar, Rahul; Mohan, Navinath; Upadhyay, Ashish Datt; Singh, Amrendra Pratap; Sahu, Vishal; Dwivedi, Sadanand; Dey, Aparajit B; Dey, Sharmistha

2014-12-01

269

Daytime napping, sleep duration and serum C reactive protein: a population-based cohort study  

PubMed Central

Objectives To explore whether daytime napping and sleep duration are linked to serum C reactive protein (CRP), a pro-inflammatory marker, in an older aged British population. Design Cross-sectional study. Setting European Prospective Investigation into Cancer and Nutrition (EPIC)-Norfolk study. Participants A total of 5018 men and women aged 48–92?years reported their sleep habits and had serum CRP levels measured. Outcome and measures CRP was measured (mg/L) during 2006–2011 in fresh blood samples using high-sensitivity methods. Participants reported napping habits during 2002–2004, and reported sleep quantity during 2006–2007. Multivariable linear regression models were used to examine the association between napping and log-transformed CRP, and geometric mean CRP levels were calculated. Results After adjustment for age and sex, those who reported napping had 10% higher CRP levels compared with those not napping. The association was attenuated but remained borderline significant (?=0.05 (95% CI 0.00 to 0.10)) after further adjustment for social class, education, marital status, body mass index, physical activity, smoking, alcohol intake, self-reported health, pre-existing diseases, systolic blood pressure, hypnotic drug use, depression and in women-only hormone replacement therapy use. The geometric means (95% CI) of CRP levels were 2.38 (2.29 to 2.47) mg/L and 2.26 (2.21 to 2.32) mg/L for those who reported napping and no napping, respectively. A U-shaped association was observed between time spent in bed at night and CRP levels, and nighttime sleep duration was not associated with serum CRP levels. The association between napping and CRP was stronger for older participants, and among extremes of time spent in bed at night. Conclusions Daytime napping was associated with increased CRP levels in an older aged British population. Further studies are needed to determine whether daytime napping is a cause for systemic inflammation, or if it is a symptom or consequence of underlying health problems. PMID:25387759

Leng, Yue; Ahmadi-Abhari, Sara; Wainwright, Nick W J; Cappuccio, Francesco P; Surtees, Paul G; Luben, Robert; Brayne, Carol; Khaw, Kay-Tee

2014-01-01

270

[Serum antioxidative activity].  

PubMed

Model systems used in the determination of serum antioxidative activity (AOA), which differ both in the way of generating free radicals and in the mode of their detection, are clinically analyzed. The specific features and potentialities of the model systems developed at the authors' laboratory are characterized. These included yolk lipoprotein suspensions, liposomal suspensions formed from total phospholipid fraction, the hemoglobin-hydrogen peroxide-luminol system. The investigations show that most model systems for determining serum AOA contribute to the water soluble interceptors of free radicals (ascorbate, urate, plasma proteins, etc.), chelating and oxidative agents of catalytically active Fe2+ (ceruloplasmin, transferrin, albumin, etc.). The serum AOA levels measured with different model systems vary with the body's status. To determine serum AOA and the contribution of major endogenous antioxidants and inhibitors of free radical reactions may be a basis for the goal-oriented use of exogenous antioxidants in the therapy of a great variety of diseases. PMID:10204018

Klebanov, G I; Teselkin, Iu O; Babenkova, I V; Liubitski?, O B; Vladimirov, Iu A

1999-01-01

271

Multiplexed Activity-based Protein Profiling of the Human Pathogen Aspergillus fumigatus Reveals Large Functional Changes upon Exposure to Human Serum*  

PubMed Central

Environmental adaptability is critical for survival of the fungal human pathogen Aspergillus fumigatus in the immunocompromised host lung. We hypothesized that exposure of the fungal pathogen to human serum would lead to significant alterations to the organism's physiology, including metabolic activity and stress response. Shifts in functional pathway and corresponding enzyme reactivity of A. fumigatus upon exposure to the human host may represent much needed prognostic indicators of fungal infection. To address this, we employed a multiplexed activity-based protein profiling (ABPP) approach coupled to quantitative mass spectrometry-based proteomics to measure broad enzyme reactivity of the fungus cultured with and without human serum. ABPP showed a shift from aerobic respiration to ethanol fermentation and utilization over time in the presence of human serum, which was not observed in serum-free culture. Our approach provides direct insight into this pathogen's ability to survive, adapt, and proliferate. Additionally, our multiplexed ABPP approach captured a broad swath of enzyme reactivity and functional pathways and provides a method for rapid assessment of the A. fumigatus response to external stimuli. PMID:22865858

Wiedner, Susan D.; Burnum, Kristin E.; Pederson, LeeAnna M.; Anderson, Lindsey N.; Fortuin, Suereta; Chauvigné-Hines, Lacie M.; Shukla, Anil K.; Ansong, Charles; Panisko, Ellen A.; Smith, Richard D.; Wright, Aaron T.

2012-01-01

272

Multiplexed activity-based protein profiling of the human pathogen Aspergillus fumigatus reveals large functional changes upon exposure to human serum.  

PubMed

Environmental adaptability is critical for survival of the fungal human pathogen Aspergillus fumigatus in the immunocompromised host lung. We hypothesized that exposure of the fungal pathogen to human serum would lead to significant alterations to the organism's physiology, including metabolic activity and stress response. Shifts in functional pathway and corresponding enzyme reactivity of A. fumigatus upon exposure to the human host may represent much needed prognostic indicators of fungal infection. To address this, we employed a multiplexed activity-based protein profiling (ABPP) approach coupled to quantitative mass spectrometry-based proteomics to measure broad enzyme reactivity of the fungus cultured with and without human serum. ABPP showed a shift from aerobic respiration to ethanol fermentation and utilization over time in the presence of human serum, which was not observed in serum-free culture. Our approach provides direct insight into this pathogen's ability to survive, adapt, and proliferate. Additionally, our multiplexed ABPP approach captured a broad swath of enzyme reactivity and functional pathways and provides a method for rapid assessment of the A. fumigatus response to external stimuli. PMID:22865858

Wiedner, Susan D; Burnum, Kristin E; Pederson, LeeAnna M; Anderson, Lindsey N; Fortuin, Suereta; Chauvigné-Hines, Lacie M; Shukla, Anil K; Ansong, Charles; Panisko, Ellen A; Smith, Richard D; Wright, Aaron T

2012-09-28

273

Composition, red blood cell uptake, and serum protein binding of phytoestrogens extracted from commercial kudzu-root and soy preparations.  

PubMed

Kudzu-root and soy phytoestrogens have been associated with anti-cancer and anti-intoxication activities. Sales of capsules containing kudzu-root and soy extracts through health food stores and the Internet are unregulated. To compare efficacy, the amount of phytoestrogens present in commercial preparations and their fate in biological samples must be determined. In this study, the levels and composition of phytoestrogens in kudzu-root and soy extracts were studied using high-performance liquid chromatography with ultraviolet light detection. The bioavailability of phytoestrogens was studied by measuring red blood cell (RBC) uptake and serum protein binding ability. Phytoestrogen levels in acidified kudzu-root samples were 5- to 10-fold greater than those in nonacidified samples. Puerarin accounted for 80% of total phytoestrogens in kudzu-root. In soy extract, puerarin was absent while genistin, glycetein, and daidzin or daidzein were the major phytoestrogens. The RBC uptake depended on the phytoestrogen's polarity and molecular length. When serum was dialyzed with phytoestrogen standards in a buffer, the protein binding of phytoestrogens correlated negatively with their polarity. However, when serum was dialyzed with kudzu-root or soy extract, almost all of the phytoestrogens present in the extract bound to serum protein. Therefore, this study suggests differences in the bioavailability of phytoestrogens when they are ingested as purified compounds compared with crude plant extract. The differential composition of phytoestrogens in kudzu-root and soy may account for the differences in their therapeutic activities. PMID:12495583

Benlhabib, Elhabib; Baker, John I; Keyler, Daniel E; Singh, Ashok K

2002-01-01

274

Immune reactivity of Brucella melitensis–vaccinated rabbit serum with recombinant Omp31 and DnaK proteins  

PubMed Central

Background and objectives Brucella melitensis infection is still a major health problem for human and cattle in developing countries and the Middle East. Materials and Methods In this study, in order to screen immunogenic candidate antigens for the development of a Brucella subunit vaccine, a cytoplasmic protein (DnaK) and an outer membrane protein (Omp31) of B. melitensis were cloned, expressed in E.coli BL21 and then purified using Ni-NTA agarose. Immunized serum was prepared from a rabbit inoculated with attenuated B. melitensis. Results and Conclusion It was proved that immunized serum contains antibodies against recombinant Omp31 (rOmp31) and DnaK (rDnaK) by Western blot and ELISA assays. The results may suggest the importance of these proteins as subunit vaccines against B. melitensis as well as targets for immunotherapy. PMID:23467315

Ghasemi, Amir; Salari, Mohammad Hossein; Zarnani, Amir Hassan; Pourmand, Mohammad Reza; Ahmadi, Hojat; Mirshafiey, Abbas; Jeddi-Tehrani, Mahmood

2013-01-01

275

Polymorphisms in the human surfactant protein-D ( SFTPD ) gene: strong evidence that serum levels of surfactant protein-D (SPD) are genetically influenced  

Microsoft Academic Search

The collectin surfactant protein-D (SP-D) plays a significant role in innate immunity. Epidemiological studies described associations between single nucleotide polymorphisms (SNPs) of the human gene coding surfactant protein-D (SFTPD) and infectious pulmonary diseases. Studies on twins indicated very strong genetic dependence for serum levels of SP-D. The aim of this study was to determine the genetic influence of sequence variations

Kathrin Heidinger; Inke R. König; Anette Bohnert; Anja Kleinsteiber; Anne Hilgendorff; Ludwig Gortner; Andreas Ziegler; Trinad Chakraborty; Gregor Bein

2005-01-01

276

Serum concentrations of insulin-like growth factor-binding protein 5 in Crohn’s disease  

PubMed Central

AIM: To investigate serum insulin-like growth factor-binding protein 5 (IGFBP-5) levels and intestinal IGFBP-5 expression in patients with Crohn’s disease (CD). METHODS: We analyzed the serum concentrations and intestinal expression of IGFBP-5 in 42 patients with CD, of whom 26 had endoscopically or radiologically proven stricture formation. Nine of the 42 patients had active disease, with a Crohn’s disease activity index > 150. Serum IGFBP-5 levels were analyzed in 20 healthy controls matched by sex and age to the CD patients. Serum IGFBP-5 was measured using an enzyme-linked immunosorbent assay. Intestinal tissue was obtained from patients through endoscopic biopsies. IGFBP-5 expression was detected using immunohistochemistry and was scored semiquantitatively. RESULTS: The median serum IGFBP-5 concentrations of CD patients were significantly lower compared with healthy controls [median 7.2 (IQR: 5.5-11.3) ng/mL vs 11.3 (8.0-44.6) ng/mL, P < 0.001]. There was no significant difference between median serum IGFBP-5 levels in CD patients with or without stricture formation [6.9 (5.5-11.3) ng/mL vs 7.8 (5.3-10.1) ng/mL, P = 0.815]. The serum IGFBP-5 levels were not significantly different between patients with active disease and inactive disease [7.2 (6.5-7.6) ng/mL vs 7.2 (5.5-11.3) ng/mL, P = 0.890]. However, a significant correlation was observed between serum IGFBP-5 levels and platelet count (PLT) (r = 0.319, P = 0.0395). No significant correlation was found between tissue IGFBP-5 immunohistochemical staining intensity scores and serum IGFBP-5 levels. No significant difference was found when comparing the serum IGFBP-5 levels among the patients with different tissue IGFBP-5 staining scores (absent/very weak, weak, moderate or strong). There was a significant correlation between tissue IGFBP-5 staining scores and white blood cell count (r = 0.391, P = 0.01) and PLT (r = 0.356, P = 0.021). CONCLUSION: Our results indicate that serum IGFBP-5 concentrations were lower in CD patients compared to healthy controls regardless of disease activity or the presence of stricture formation. PMID:24379630

Adali, Gupse; Yorulmaz, Elif; Ozkanli, Seyma; Ulasoglu, Celal; Bayraktar, Baris; Orhun, Alev; Colak, Yasar; Tuncer, Ilyas

2013-01-01

277

Serum bone Gla protein and carboxyterminal cross-linked telopeptide of type I collagen in patients with Cushing's syndrome.  

PubMed Central

Serum bone Gla protein, a marker of bone formation, and carboxyterminal cross-linked telopeptide of type I collagen levels, an index of bone resorption, were evaluated in eight patients with active Cushing's syndrome and in four with 'preclinical' Cushing's syndrome, before and after surgery. In basal conditions, serum bone Gla protein levels were significantly lower (p < 0.0001) in patients with active Cushing's syndrome (1.0 +/- 0.35 ng/ml) than in controls (5.4 +/- 0.15 ng/ml); two out of four patients with the 'preclinical' form had reduced bone Gla protein levels, while in the other two cases levels were in the normal range. Serum levels of carboxyterminal cross-linked telopeptide of type I collagen (3.0 +/- 0.4 ng/ml), although slightly reduced, were similar to those recorded in controls (4.1 +/- 0.3 ng ml), both in patients with active and with preclinical Cushing's syndrome. After surgery serum levels of both marker proteins significantly increased in seven out of eight patients with active Cushing's syndrome; in one patient, who was not cured after surgery, bone Gla proteins levels remained lower than in normals, while levels of carboxyterminal cross-linked telopeptide of type I collagen had a transient increase after six months. In the two patients with a 'preclinical' Cushing's syndrome who underwent surgery, a significant rise of the levels of both marker proteins was observed, similar to that observed in patients with active Cushing's syndrome. It was concluded that serial determinations of these new markers of bone formation and resorption may be usefully employed to follow-up the clinical course of Cushing's syndrome and provide information on the rate of bone turnover in response to medical and/or surgical therapies. Moreover, the evaluation of these markers in preclinical states of Cushing's syndrome might suggest the need for surgery. PMID:8935602

Sartorio, A.; Conti, A.; Ferrario, S.; Passini, E.; Re, T.; Ambrosi, B.

1996-01-01

278

Proteomics of colorectal cancer: overview of discovery studies and identification of commonly identified cancer-associated proteins and candidate CRC serum markers.  

PubMed

Colorectal cancer (CRC) is a common cause of cancer-related mortality in the developed world. Improved methods for early detection and disease management are urgently needed. Many efforts in the past 5 years have been devoted to protein biomarker discovery for early detection of CRC. Here, we discuss identity-based studies employing tandem mass spectrometry that analyzed clinical material as well as model systems. Through meta-analysis we provide a list of CRC-associated tissue proteins discovered in multiple studies, with the greater majority being 2D gel-based discoveries coupled to MS/MS. So far only a limited number of CRC-associated proteins have been validated in serum for non-invasive testing for CRC. This list includes several intracellular and nuclear proteins that a priori would not have been considered candidate biomarkers based on their predicted subcellular localization. Finally, we highlight promising new directions that combine targeted analyses of subcellular proteomes, like the cell surface, secretome, exosome, and nuclear matrix, with nanoLC-MS/MS-based proteomics. We anticipate that in the near future, these novel mass spectrometry-based in-depth approaches will uncover many novel, specific CRC marker candidates in clinical tissues and that their targeted validation with multi-reaction monitoring MS will speed up development of non-invasive tests in feces and serum/plasma. PMID:20601272

Jimenez, Connie R; Knol, Jaco C; Meijer, Gerrit A; Fijneman, Remond J A

2010-09-10

279

Impact of scaling and root planing on C-reactive protein levels in gingival crevicular fluid and serum in chronic periodontitis patients with or without diabetes mellitus  

PubMed Central

Purpose The present study was conducted to evaluate the impact of scaling and root planing (SRP) on the C-reactive protein (CRP) levels of gingival crevicular fluid (GCF) and serum in chronic periodontitis patients with type 2 diabetes mellitus (T2DM-CP) or without type 2 diabetes mellitus (NDM-CP). Methods Forty-eight human participants were divided into two groups: an experimental (T2DM-CP) group (group I, n=24) comprising chronic periodontitis patients with random blood sugar ?200 mg/dL and type 2 diabetes mellitus, and control (NDM-CP) group (group II, n=24) of those with chronic periodontitis and random blood sugar <200 without T2DM for the study. All subjects underwent nonsurgical periodontal therapy (NSPT) including complete SRP and subgingival debridement. Periodontal health parameters, plaque index (PI), gingival index (GI), probing pocket depth (PPD), clinical attachment level (CAL), GCF volume (GCF vol), GCF-CRP, random blood glucose (RBS), glycated hemoglobin, and systemic inflammatory markers, serum CRP, total leukocyte count (TLC), neutrophil count (Neutr) and lymphocyte count (Lymph), were evaluated at baseline, 1 month, and 3 months after SRP. Results NSPT resulted in statistically significant improvement in periodontal health parameters (PI, GI, PPD, CAL, GCF vol), CRP levels in serum as well as GCF of both groups I and II. The mean improvement in periodontal health parameters (PI, GI, PPD, CAL, GCF vol), CRP levels in serum and GCF was greater in group I than group II after NSPT. There was nonsignificant increase in GCF-CRP, TLC, Lymph, and RBS, and a significant increase in Neutr and Serum CRP in group II at 1 month. The Serum CRP level of 20 out of 24 group II patients had also increased at 1 month. Conclusions The CRP levels in both GCF and serum were higher in T2DM-CP patients than in NDM-CP patients. Although there was a significant improvement in both the groups, greater improvement was observed in both GCF and serum samples of T2DM-CP patients. Graphical Abstract PMID:25177517

2014-01-01

280

Internally calibrated quantification of protein analytes in human serum by fluorescence immunoassays in disposable elastomeric microfluidic devices  

PubMed Central

Herein we report on reliable reproducible quantification of protein analytes in human serum by fluorescence sandwich immunoassays in disposable PDMS microfluidic chips. The system requires 1000 times less sample than typical clinical blood tests and is specifically shown to measure ferritin down to 250 pM in human serum. The in-built calibration method of spiking the serum with known concentrations of commercially available antigen avoids common sources of error and improves the reliability of the test results. The reported microfluidic system is an important new tool for fundamental scientific research, offering sensitive immunoassay measurements in small but complex biosamples. The system is also a further step towards comprehensive affordable “point-of-care” biomedical diagnostics. PMID:19130581

Kartalov, Emil P.; Lin, David H.; Lee, David T.; Anderson, William F.; Taylor, Clive R.; Scherer, Axel

2009-01-01

281

The elevation of serum napsin A in idiopathic pulmonary fibrosis, compared with KL-6, surfactant protein-A and surfactant protein-D  

PubMed Central

Background Napsin A, an aspartic protease, is mainly expressed in alveolar type-II cells and renal proximal tubules and is a putative immunohistochemical marker for pulmonary adenocarcinomas. This study sought to determine whether napsin A could be measured in the serum to evaluate its relationship to idiopathic pulmonary fibrosis (IPF) and determine whether renal dysfunction might affect serum napsin A levels. Methods Serum levels of napsin A were measured in 20 patients with IPF, 34 patients with lung primary adenocarcinoma, 12 patients with kidney diseases, and 20 healthy volunteers. Surfactant protein (SP)-A, SP-D, and Krebs von den Lungen-6 (KL-6) levels in serum and pulmonary function tests were also evaluated in IPF patients. Results Circulating levels of napsin A were increased in patients with IPF, as compared with healthy controls, and they correlated with the severity of disease. Moreover, the serum napsin A levels were not elevated in patients with pulmonary adenocarcinoma or renal dysfunction. The distinguishing point between IPF and the controls was that the area under the receiver operating characteristic curve (ROC) of napsin A was larger than that of KL-6, SP-A, or SP-D. Conclusion These findings suggest that serum napsin A may be a candidate biomarker for IPF. PMID:22963039

2012-01-01

282

Development of a "membrane cloaking" method for amperometric enzyme immunoassay and surface plasmon resonance analysis of proteins in serum samples.  

PubMed

Detection of trace amounts of target proteins in the presence of high concentrations of matrix proteins (e.g., serum samples) without separation steps is of great significance to biomedical research but remains technically challenging. Here we report a "membrane cloaking" method to overcome nonspecific protein adsorption and fouling problems for label-free surface plasmon resonance detection and heterogeneous immunosensing. A thin, hybrid, self-assembled monolayer on gold was formed with 70 mol % mercaptopropanol and 30 mol % cysteamine/propanedithiol to facilitate membrane fusion and covalent attachment of antibodies. After antibody immobilization, the surface was incubated with lipid vesicles, which fused to form a supported membrane. The analyte spiked in serum was introduced for binding, and the membrane and nonspecifically adsorbed proteins on the membrane were subsequently removed using a nonionic surfactant before the final measurement was carried out. Selection of a suitable surfactant can preserve antibody/antigen binding and selectively remove the membrane, allowing accurate measurement of the captured proteins without interference from nonspecifically adsorbed species. Surface plasmon resonance (SPR) quantification of IgG spiked in undiluted serum ( approximately 75 mg/mL protein) was achieved with the membrane cloaking method, whereas direct measurement without membrane removal resulted in a significantly large error. The cloaking method was also used to develop an enzyme amplified amperometric assay using HRP-conjugated IgG. Detection of concentrations as low as 5 fM proteins was obtained. Finally, a membrane cloaking assay combining SPR and in situ electrochemical measurement was demonstrated on a gold substrate. Similar sensitivity was observed using a continuous flow injection measurement. The method opens new avenues to develop direct assay methods with ultrahigh sensitivity for protein samples using SPR and enzyme-linked amplification mechanisms. PMID:17263314

Phillips, K Scott; Han, Jong Ho; Cheng, Quan

2007-02-01

283

In vitro and in vivo interactions of selected nanoparticles with rodent serum proteins and their consequences in biokinetics  

PubMed Central

Summary When particles incorporated within a mammalian organism come into contact with body fluids they will bind to soluble proteins or those within cellular membranes forming what is called a protein corona. This binding process is very complex and highly dynamic due to the plethora of proteins with different affinities and fractions in different body fluids and the large variation of compounds and structures of the particle surface. Interestingly, in the case of nanoparticles (NP) this protein corona is well suited to provide a guiding vehicle of translocation within body fluids and across membranes. This NP translocation may subsequently lead to accumulation in various organs and tissues and their respective cell types that are not expected to accumulate such tiny foreign bodies. Because of this unprecedented NP accumulation, potentially adverse biological responses in tissues and cells cannot be neglected a priori but require thorough investigations. Therefore, we studied the interactions and protein binding kinetics of blood serum proteins with a number of engineered NP as a function of their physicochemical properties. Here we show by in vitro incubation tests that the binding capacity of different engineered NP (polystyrene, elemental carbon) for selected serum proteins depends strongly on the NP size and the properties of engineered surface modifications. In the following attempt, we studied systematically the effect of the size (5, 15, 80 nm) of gold spheres (AuNP), surface-modified with the same ionic ligand; as well as 5 nm AuNP with five different surface modifications on the binding to serum proteins by using proteomics analyses. We found that the binding of numerous serum proteins depended strongly on the physicochemical properties of the AuNP. These in vitro results helped us substantially in the interpretation of our numerous in vivo biokinetics studies performed in rodents using the same NP. These had shown that not only the physicochemical properties determined the AuNP translocation from the organ of intake towards blood circulation and subsequent accumulation in secondary organs and tissues but also the the transport across organ membranes depended on the route of AuNP application. Our in vitro protein binding studies support the notion that the observed differences in in vivo biokinetics are mediated by the NP protein corona and its dynamical change during AuNP translocation in fluids and across membranes within the organism. PMID:25383281

Fertsch-Gapp, Stefanie; Schäffler, Martin; Johnston, Blair D; Haberl, Nadine; Pfeiffer, Christian; Diendorf, Jörg; Schleh, Carsten; Hirn, Stephanie; Semmler-Behnke, Manuela; Epple, Matthias; Parak, Wolfgang J

2014-01-01

284

Changes of acute-phase protein levels in the serum of lung cancer patients following radiotherapy  

PubMed Central

Purpose: the assessment of serum level changes of C-reactive protein (CRP), ferritin (FER), and albumin (ALB) as inflammation markers in Non Small Cell Lung Cancer patients (stages IIIA - inoperable and stage IIIB) treated with radiotherapy. Significant findings: Normal pre-radiotherapy levels of CRP were found in 18 patients, of FER in 17, and of ALB in 22. Higher levels of CRP were found in 9 patients and of FER in 10. Lower ALB was found in 5 patients.Post-radiotherapy CRP levels were significantly higher (compared to the pre-radiotherapy levels) in 25 patients. The same was observed regarding FER in 18 patients whereas 12 patients had lower post-radiotherapy levels. The statistical analysis (non parametrical Wilcoxon test) revealed that these differences were statistically significant (p-value< 0.001). Conclusions: The levels of CRP, FER, and ALB are reliable and useful biomarkers correlated with the acute complication of lung parenchyma damage induced by radiotherapy. PMID:23236558

Maria, Tolia G; Vasileios, Kouloulias E; Panagiotis, Pantelakos S; Kostas, Syrigos N

2013-01-01

285

Serum Cartilage Oligomeric Matrix Protein: is There a Repeated Bout Effect?  

PubMed Central

The primary aim of the present study was to investigate if there is a repeated bout effect for cartilage tissue, evident in the marker serum cartilage oligomeric matrix protein (sCOMP). Ten healthy male subjects (26.4±3.14 years) performed two high impact interventions (100 drop jumps with a 30 second interval) carried out at a 3 week interval. After each intervention, sCOMP and muscle soreness were assessed on 8 and 6 occasions respectively. Muscle soreness was determined via a visual analog scale with a maximum pain score of 10. sComp levels did not show a blunted response after the second bout (Bout 1: 12.2±3.3 U/L?1; Bout 2: 13.1±4.0 U/L?1; P>0.05). Remarkably, sCOMP increased from baseline levels by 16% after bout 1 and 15% after bout 2. Muscle soreness was blunted following the second intervention (Bout 1: 5.0±1.8; Bout 2: 1.6±0.8). Unlike the known repeated bout effect for muscle damage markers, sCOMP levels do not show a blunted response after two similar loading interventions. This information on biomarker behavior is essential to clinicians attempting to use this marker as an indicator of cartilage damage associated with the development or progression of osteoarthritis. PMID:25317315

Montag, Johannes; Kilian, Yvonne; McCourt, Molly; Liphardt, Anna-Maria; Mester, Joachim

2014-01-01

286

Chemical structure of the arabinogalactan protein from gum ghatti and its interaction with bovine serum albumin.  

PubMed

Exudate gums, because of their beneficial properties, have been significant items of international trade in various industries for centuries. This manuscript sets out to gain insight into the fine structural details of an arabinogalactan protein (AGP) of gum ghatti (Anogeissus latifolia gum). The presence of a highly branched 554 kDa AGP having 1,6-linked Galp, 1,2-linked Manp, 1,3-linked Araf and 1,4-linked GlcpA main chain, substituted at O-4,6 of 1,2-linked Manp, and O-3/O-3,4 of 1,6-linked Galp residues by Araf, Arap and Galp units was revealed by chemical, chromatographic, ESMS, and NMR analyses. In particular, ESMS analysis of per acetylated oligomeric fragments derived from AGP by Smith degradation followed by acetylation was described as a commanding tool for providing critical structural information on a spectrum of glycerol tagged oligosaccharides. In addition, formation of an electrostatically driven complex between the isolated AGP and bovine serum albumin resulting in changes in the microenvironment around the tryptophan residues of BSA was established. A moderate radical scavenging activity comparable with those of standard antioxidants was observed from the AGP fraction (?94% at 1 mg/mL) that could be valuable in foods or pharmaceutical products as alternatives to synthetic antioxidants. PMID:25498648

Ghosh, Kanika; Ray, Sayani; Ghosh, Debjani; Ray, Bimalendu

2015-03-01

287

Factors Influencing the Measurement of Plasma/Serum Surfactant Protein D Levels by ELISA  

PubMed Central

Background Extensive variations in human surfactant protein D (SP-D) levels in circulation as measured by ELISA exist in the published literature. In order to determine the source of these variations, factors influencing the measurement by ELISA were explored. Materials and Methods Peripheral blood from healthy individuals was collected into various vacutainers during the same blood draw. Recombinant SP-D was diluted into different matrices and used for a standard curve. Samples were analyzed by capture ELISA using one of two distinct detection antibodies. Results The type of matrix had some effects on detection of recombinant SP-D. The type of anticoagulant used and dilution factor had very little effect, except for in plasma collected in EDTA vacutainers. The extent of variation in published values seemed to be due to the ELISA configuration employed, and, in agreement with this, we found that by switching the detection antibody, there was a 50% decrease in the extrapolated SP-D value of serum and plasma samples. Storage of samples resulted in slight changes in measured SP-D levels. Conclusions The ELISA configuration employed to measure circulating levels of SP-D has a significant effect on the extrapolated values. In both configurations tested, the use of EDTA as a coagulant resulted in inconsistent values, and we, therefore, suggest the avoidance of this anticoagulant when assaying for SP-D by ELISA. While the demonstrated effects of several factors on measurement of SP-D may not account for all the disparities amongst the previous studies, they stress that variations in methodologies for measuring the same protein can result in very inconsistent results. PMID:25365324

Bratcher, Preston E.; Gaggar, Amit

2014-01-01

288

Protective Molecules–C-Reactive Protein (CRP), Serum Amyloid P (SAP), Pentraxin3 (PTX3), Mannose-Binding Lectin (MBL), and Apolipoprotein A1 (Apo A1), and Their Autoantibodies: Prevalence and Clinical Significance in Autoimmunity  

Microsoft Academic Search

Apoptotic defects and impaired clearance of cellular debris are considered key events in the development of autoimmunity,\\u000a as they can contribute to autoantigen overload, and may initiate an autoimmune response. The pentraxins are a group of highly\\u000a conserved proteins including the short pentraxins, C-reactive protein (CRP) and serum amyloid-P (SAP), and the long pentraxin-3\\u000a (PTX3), which are all involved in

Martine Szyper Kravitz; Milena Pitashny; Yehuda Shoenfeld

2005-01-01

289

Direct detection of C-reactive proteins in human serum using nanoparticle-enhanced surface plasmon resonance biosensing  

NASA Astrophysics Data System (ADS)

C-reactive protein (CRP) produced by the liver is one of the most characteristic acute-phase proteins. It has been suggested that the level of CRP in human serum may be a significant tool of detecting risks of developing cardiovascular disease and atherosclerosis. Here we propose an advanced plasmonic surface plasmon resonance (SPR) bioassay with Au nanoparticles embedded in the dielectric film that demonstrates a 10X improvement in resolution compared to the conventional SPR biosensor. The co-sputtered film was modified with (3-Aminopropyl)triethoxysilane to sequentially immobilize protein G, monoclonal anti-CRP antibody (C8), and human serum albumins (HSA). After blocked by ethanolamine, the sensor was used to detect CRP. Using this extremely sensitive biochip, the lowest reliable concentration of CRP without any exterior labeling is simplified to human physiological level. The novel assay has the latent capability of not only eliminating the disturbances coming from serum proteins resulting in false signals, but is also able to be applied in rapid and label-free clinical detections of CRP with large improved sensitivity.

Lin, H.-Y.; Tsang, K. Y.; Hu, W. P.; Hsu, H.-Y.; Chiou, A.; Chang, G.-L.; Chen, S.-J.

2006-08-01

290

Correlation between Serum Levels of High Mobility Group Box-1 Protein and Pancreatitis: A Meta-Analysis  

PubMed Central

Background. Aberrant expression of high mobility group box-1 protein (HMGB1) contributes to the progression of various inflammatory diseases. This meta-analysis focused on the clinical significance of serum HMGB1 levels in pancreatitis patients, with the goal of building a novel diagnostic score model. Method. We conducted a meta-analysis by searching in the PubMed, Embase, Web of Science, Cochrane Library, CISCOM, CINAHL, Google Scholar, China BioMedicine (CBM), and China National Knowledge Infrastructure (CNKI) databases without any language restrictions. Studies were pooled and standard mean difference (SMD) and its corresponding 95% confidence intervals (95% CIs) were calculated. Version 12.0 STATA software was used for statistical analysis. Results. We performed a final analysis of 841 subjects from 12 clinical case-control studies. The meta-analysis results showed a positive association between serum HMGB1 levels and the progression of pancreatitis. In the subgroup analysis by country, high serum level of HMGB1 may be related to pancreatitis progression in China, Korea, Hungary, and Japan populations (all P < 0.05). Conclusion. The present meta-analysis indicated that serum HMGB1 level was statistically elevated in patients with pancreatitis, and thus serum levels of HMGB1 could be determined to be a useful biomarker for pancreatitis patients. PMID:25695079

Lin, Yan; Lin, Lian-Jie; Jin, Yu; Cao, Yong; Zhang, Ying; Zheng, Chang-Qing; Liu, Jia-Li; Yang, Sheng-Li

2015-01-01

291

Serum cleaved Tau protein and neurobehavioral battery of tests as markers of brain injury in experimental bacterial meningitis.  

PubMed

Brain injury due to bacterial meningitis affects multiple areas of the brain with a heterogeneous distribution generating a challenge to assess severity. Tau proteins are microtubular binding proteins localized in the axonal compartment of neurons. Brain injury releases cleaved Tau proteins (C-tau) into the extracellular space where they are transported to the cerebral spinal fluid. We hypothesized that C-tau crosses the blood-brain barrier during inflammation and that it can be detected in serum. The correlation between serum C-tau levels and the extent of the meningitic insult was examined. Furthermore, we studied whether the use of a subset of neurobehavioral tasks can assess the extent of brain injury after meningitis. The tests were chosen primarily for their ability to detect deficits in the acoustic system, low brain, reflexive responding, as well as for impaired motor coordination and the higher brain functions of learning and memory. A rat model of group B streptococcal meningitis with variable severity was utilized. At five days after bacterial inoculation followed by antibiotic therapy neurobehavioral tests were performed and serum C-tau and histologic samples of the brain were obtained. Our study shows that during meningitis C-tau appears in serum and reflects the extent of neurologic damage. Neurobehavioral performance was altered after bacterial meningitis and could be correlated with histologic and biochemical markers of neurologic sequelae. We conclude that serum C-tau and a composite of neurobehavioral tests could become useful markers for assessing the severity of neurological damage in experimental bacterial meningitis. PMID:11532253

Irazuzta, J E; de Courten-Myers, G; Zemlan, F P; Bekkedal, M Y; Rossi, J

2001-09-14

292

X-ray absorption spectroscopy studies of the adducts formed between cytotoxic gold compounds and two major serum proteins.  

PubMed

Gold metallodrugs form a class of promising antiproliferative agents showing a high propensity to react with proteins. We exploit here X-ray absorption spectroscopy (XAS) methods [both X-ray absorption near-edge spectroscopy (XANES) and extended X-ray absorption fine structure (EXAFS)] to gain insight into the nature of the adducts formed between three representative gold(I, III) metallodrugs (i.e., auranofin, [Au(2,2'-bipyridine)(OH)(2)](PF(6)), Aubipy, and dinuclear [Au(2)(6,6'-dimethyl-2,2'-bipyridine)(2)(?-O)(2)](PF(6))(2), Auoxo6) and two major plasma proteins, namely, bovine serum albumin (BSA) and human serum apotransferrin (apoTf). The following metallodrug-protein systems were investigated in depth: auranofin/apoTf, Aubipy/BSA, and Auoxo6/apoTf. XANES spectra revealed that auranofin, upon protein binding, conserves its gold(I) oxidation state. Protein binding most probably takes place through release of the thiosugar ligand and its subsequent replacement by a thiol (or a thioether) from the protein. This hypothesis is independently supported by EXAFS results. In contrast, the reactions of Aubipy with serum albumin and of Auoxo6 with serum apoTf invariantly result in gold(III) to gold(I) reduction. Gold(III) reduction, clearly documented by XANES, is accompanied, in both cases, by release of the bipyridyl ligands; for Auoxo6 cleavage of the gold-gold dioxo bridge is also observed. Gold(III) reduction leads to formation of protein-bound gold(I) species, with deeply modified metal coordination environments, as evidenced by EXAFS. In these adducts, the gold(I) centers are probably anchored to the protein through nitrogen donors. In general, these two XAS methods, i.e., XANES and EXAFS, used here jointly, allowed us to gain independent structural information on metallodrug/protein systems; detailed insight into the gold oxidation state and the local environment of protein-bound metal atoms was achieved in the various cases. PMID:21181484

Messori, L; Balerna, A; Ascone, I; Castellano, C; Gabbiani, C; Casini, A; Marchioni, C; Jaouen, G; Congiu Castellano, A

2011-03-01

293

Reduction of protein concentration range difference followed by multicolumn fractionation prior to 2-DE and LC-MS/MS profiling of serum proteins.  

PubMed

This article is concerned with the reduction of protein concentration range differences by the peptide beads library technology (ProteoMiner™ or "equalizer" technology), which in principle allows the enrichment of proteins to the same concentration level (i.e. protein equalizer) regardless of the original protein abundance in a given biological fluid such as human serum, which is the subject of our investigation. After the equalization step, the captured proteins from human serum were fractionated on a series of tandem monolithic columns with surface-bound iminodiacetic acid ligands to which three different metal ions, namely, Zn²+, Ni²+ and Cu²+ were immobilized to yield the so-called immobilized metal affinity chromatography columns. These three monolithic columns were connected to a reversed-phase column packed with polystyrene divinyl benzene beads. Aliquots taken from the four collected fractions from the four tandem columns were subsequently fractionated by 2-DE. Also, aliquots from the four collected fractions were tryptically digested and analyzed by LC-MS/MS. The strategy of subsequent fractionation on the four tandem columns after equalization allowed the identification of more proteins than simply using the equalization by ProteoMiner™ . The equalizer technology was compared to the immuno-subtraction approach. While the ProteoMiner™ technology is superior in terms of the overall number of captured proteins, it only complements the immuno-subtraction approach since the latter can capture the proteins that were not captured by the former. PMID:21365658

Selvaraju, Subhashini; El Rassi, Ziad

2011-03-01

294

The influence of naturally occurring heterophilic anti-immunoglobulin antibodies on direct measurement of serum proteins using sandwich ELISAs.  

PubMed

Sandwich ELISAs have become a widely used method for the quantitative detection of serum proteins. However, they can be biased by a variety of interfering substances. As reported recently, we observed false-positive levels of interferon (IFN)-alpha and -beta in up to 27% of sera from healthy blood donors using commercial ELISAs. We now demonstrate that two different groups of naturally occurring heterophilic antibodies (IgG-type) are responsible for these titers. Group I (representing 85% of positive samples) binds to the Fab region of IgG from goat, mouse, rat, horse, and bovidae (but not rabbit). Group II (15%) recognizes an epitope in the Fc region of mouse, horse, bovine, and rabbit (but not goat or rat) immunoglobulins. The antibodies did not crossreact with human IgG subclasses but contributed to false-positive IgG rheumatoid factor levels obtained using a commercially available ELISA. To investigate the susceptibility of assays to these artifacts, various combinations of capture and detection antibodies have been tested. On this basis, we defined the relative risks that standard ELISAs might be influenced by heterophilic anti-immunoglobulin antibodies. In general, assays that use monoclonal antibodies for both capture and detection are less susceptible than others which include at least one polyclonal antiserum. However, only systems utilizing rabbit F(ab')(2) fragments have been found to be immune to this interference. PMID:10675759

Hennig, C; Rink, L; Fagin, U; Jabs, W J; Kirchner, H

2000-02-21

295

Proteomic analysis of serum proteins in triple transgenic Alzheimer's disease mice: implications for identifying biomarkers for use to screen potential candidate therapeutic drugs for early Alzheimer's disease.  

PubMed

Alzheimer's disease (AD) is the most common fatal neurodegenerative disease affecting the elderly worldwide. There is an urgent need to identify novel biomarkers of early AD. This study aims to search for potential early protein biomarkers in serum from a triple transgenic (PS1M146V/APPSwe/TauP301L) mouse model. Proteomic analysis via two-dimensional fluorescence difference gel electrophoresis was performed on serum samples from wild-type (WT) and triple transgenic mice that were treated with or without coenzyme Q10 (CoQ10) (800 mg/kg body weight/day), a powerful endogenous antioxidant displaying therapeutic benefits against AD pathology and cognitive impairment in multiple AD mouse models, for a period of three months beginning at two months of age. A total of 15 differentially expressed serum proteins were identified between the WT and AD transgenic mice. The administration of CoQ10 was found to alter the changes in the differentially expressed serum proteins by upregulating 10 proteins and down-regulating 10 proteins. Among the proteins modulated by CoQ10, clusterin and ?-2-macroglobulin were validated via ELISA assay. These findings revealed significant changes in serum proteins in the AD mouse model at an early pathological stage and demonstrated that administration of CoQ10 could modulate these changes in serum proteins. Our study suggested that these differentially expressed serum proteins could serve as potential protein biomarkers of early AD and that screening for potential candidate AD therapeutic drugs and monitoring of therapeutic effects could be performed via measurement of the changes in these differentially expressed serum proteins. PMID:24496070

Sui, Xiaojing; Ren, Xiaohu; Huang, Peiwu; Li, Shuiming; Ma, Quan; Ying, Ming; Ni, Jiazuan; Liu, Jianjun; Yang, Xifei

2014-01-01

296

The renal handling of low molecular weight proteins. II. Disorders of serum protein catabolism in patients with tubular proteinuria, the nephrotic syndrome, or uremia.  

PubMed

The present study was directed toward determining the role of the kidney in the metabolism of various classes of serum proteins and to define the urinary protein excretion patterns and the pathogenesis of disorders of protein metabolism in patients with proteinuria. To this end, the metabolic fates of a small protein, lambda-L chain (mol wt 44,000), and a protein of intermediate size, IgG (mol wt 160,000), were studied in controls and patients with renal disease. Controls metabolized 0.28%/hr of circulating IgG and 22.3%/hr of circulating lambda-L chain. All the IgG and 99% of the lambda-L chain was catabolized with the remaining lambda-L chain lost intact into the urine. The kidney was shown to be the major site of catabolism for small serum proteins. Three distinct disorders of protein metabolism were noted in patients with renal tubular disease and tubular proteinuria, glomerular disease (the nephrotic syndrome), and disease involving the entire nephrons (uremia), respectively. Patients with renal tubular disease had a 50-fold increase in the daily urinary excretion of 15-40,000 molecular weight proteins such as lysozyme and lambda-L chains. Serum IgG and lambda-L chain survivals were normal; however, the fraction of the over-all lambda-L chain metabolism accounted for by proteinuria was increased 40-fold whereas endogenous catabolism was correspondingly decreased. Thus, tubular proteinuria results from a failure of proximal tubular uptake and catabolism of small proteins that are normally filtered through the glomerulus. Patients with the nephrotic syndrome had a slight increase in lambda-L chain survival whereas IgG survival was decreased and the fraction of IgG lost in the urine was markedly increased. Here, abnormal glomerular permeability to proteins of intermediate size is the basic abnormality. Patients with uremia had a normal IgG survival but a four to 10-fold prolongation of lambda-L chain survival due to loss of entire nephrons, the major site of metabolism of these proteins. This results in an increase (up to 10-fold) in the serum concentration of lambda-L chain, lysozyme, and other small biologically active proteins, a phenomenon that may be of importance in causing some of the manifestations of the uremic syndrome. PMID:5054468

Waldmann, T A; Strober, W; Mogielnicki, R P

1972-08-01

297

Fatty acid-binding site environments of serum vitamin D-binding protein and albumin are different  

PubMed Central

Vitamin D-binding protein (DBP) and albumin (ALB) are abundant serum proteins and both possess high-affinity binding for saturated and unsaturated fatty acids. However, certain differences exist. We surmised that in cases where serum albumin level is low, DBP presumably can act as a transporter of fatty acids. To explore this possibility we synthesized several alkylating derivatives of 14C-palmitic acid to probe the fatty acid binding pockets of DBP and ALB. We observed that N-ethyl-5-phenylisooxazolium-3?-sulfonate-ester (WRK ester) of 14C-palmitic acid specifically labeled DBP; but p-nitrophenyl- and N-hydroxysuccinimidyl-esters failed to do so. However, p-nitrophenyl ester of 14C-palmitic acid specifically labeled bovine ALB, indicating that the micro-environment of the fatty acid-binding domains of DBP and ALB may be different; and DBP may not replace ALB as a transporter of fatty acids. PMID:18374965

Swamy, Narasimha; Ray, Rahul

2008-01-01

298

Simple and rapid solid-phase radioimmunoassay for serum progesterone, using the protein A of Staphylococcus aureus as immunoadsorbent  

Microsoft Academic Search

A simple, rapid, and inexpensive radioimmunoassay method for serum progesterone is described, which uses a solid-phase technique for separation of antibody-bound from antibody-free progesterone. Rabbit antiprogesterone immunoglobulins are adsorbed on the protein A of formaldehyde- and heat-treated Staphylococcus aureus cells (Pansorbin; Calbiochem-Behring Corp., La Jolla, California). The suspension of antibody-coated Pansorbin retains all its binding activity of 1-2-H(N)-progesterone when kept

J. Jungers; J. Delogne-Desnoeck; C. Robyn

1981-01-01

299

Selection of symptomatic patients with Crohn's disease for abdomino-pelvic CT: Role of serum C-reactive protein  

Microsoft Academic Search

BackgroundPrevious studies have shown that repeated abdomino-pelvic computed tomography (CT) examinations can lead to substantial cumulative diagnostic radiation exposure in patients with Crohn's disease (CD). Improved selection of patients referred for CT will reduce unnecessary radiation exposure. This study determines if serum C-reactive protein (CRP) concentration predicts which symptomatic patients with CD are likely to have significant disease activity or

Alan N. Desmond; Kevin O'Regan; Neera Malik; Sebastian McWilliams; Siobhan O'Neill; Eamonn M. Quigley; Fergus Shanahan; Michael M. Maher

300

Serial Changes in Surfactant-associated Proteins in Lung and Serum before and after Onset of ARDS  

Microsoft Academic Search

The goal of this study was to determine the changes that occur in surfactant-associated proteins in bronchoalveolar lavage fluid (BAL) and serum of patients at risk for ARDS and during the course of ARDS. We found that the concentrations of SP-A and SP-B were low in the BAL of patients at risk for ARDS before the onset of clinically defined

KELLY E. GREENE; JO RAE WRIGHT; KENNETH P. STEINBERG; JOHN T. RUZINSKI; ELLEN CALDWELL; WES B. WONG; WILLIAM HULL; JEFFREY A. WHITSETT; TOYOAKI AKINO; YOSHIO KUROKI; HISATO NAGAE; LEONARD D. HUDSON; THOMAS R. MARTIN

1999-01-01

301

Label-Free LC-MSe in Tissue and Serum Reveals Protein Networks Underlying Differences between Benign and Malignant Serous Ovarian Tumors  

PubMed Central

Purpose To identify proteins and (molecular/biological) pathways associated with differences between benign and malignant epithelial ovarian tumors. Experimental Procedures Serum of six patients with a serous adenocarcinoma of the ovary was collected before treatment, with a control group consisting of six matched patients with a serous cystadenoma. In addition to the serum, homogeneous regions of cells exhibiting uniform histology were isolated from benign and cancerous tissue by laser microdissection. We subsequently employed label-free liquid chromatography tandem mass spectrometry (LC-MSe) to identify proteins in these serum and tissues samples. Analyses of differential expression between samples were performed using Bioconductor packages and in-house scripts in the statistical software package R. Hierarchical clustering and pathway enrichment analyses were performed, as well as network enrichment and interactome analysis using MetaCore. Results In total, we identified 20 and 71 proteins that were significantly differentially expressed between benign and malignant serum and tissue samples, respectively. The differentially expressed protein sets in serum and tissue largely differed with only 2 proteins in common. MetaCore network analysis, however inferred GCR-alpha and Sp1 as common transcriptional regulators. Interactome analysis highlighted 14-3-3 zeta/delta, 14-3-3 beta/alpha, Alpha-actinin 4, HSP60, and PCBP1 as critical proteins in the tumor proteome signature based on their relative overconnectivity. The data have been deposited to the ProteomeXchange with identifier PXD001084. Discussion Our analysis identified proteins with both novel and previously known associations to ovarian cancer biology. Despite the small overlap between differentially expressed protein sets in serum and tissue, APOA1 and Serotransferrin were significantly lower expressed in both serum and cancer tissue samples, suggesting a tissue-derived effect in serum. Pathway and subsequent interactome analysis also highlighted common regulators in serum and tissue samples, suggesting a yet unknown role for PCBP1 in ovarian cancer pathophysiology. PMID:25265318

Wegdam, Wouter; Argmann, Carmen A.; Kramer, Gertjan; Vissers, Johannes P.; Buist, Marrije R.; Kenter, Gemma G.; Aerts, Johannes M. F. G.; Meijer, Danielle; Moerland, Perry D.

2014-01-01

302

The Effect of Simulated Microgravity Environment of RWV Bioreactors on Surface Reactions and Adsorption of Serum Proteins on Bone-bioactive Microcarriers  

NASA Technical Reports Server (NTRS)

Biomimetically modified bioactive materials with bone-like surface properties are attractive candidates for use as microcarriers for 3-D bone-like tissue engineering under simulated microgravity conditions of NASA designed rotating wall vessel (RWV) bioreactors. The simulated microgravity environment is attainable under suitable parametric conditions of the RWV bioreactors. Ca-P containing bioactive glass (BG), whose stimulatory effect on bone cell function had been previously demonstrated, was used in the present study. BG surface modification via reactions in solution, resulting formation of bone-like minerals at the surface and adsorption of serum proteins is critical for obtaining the stimulatory effect. In this paper, we report on the major effects of simulated microgravity conditions of the RWV on the BG reactions surface reactions and protein adsorption in physiological solutions. Control tests at normal gravity were conducted at static and dynamic conditions. The study revealed that simulated microgravity remarkably enhanced reactions involved in the BG surface modification, including BG dissolution, formation of bone-like minerals at the surface and adsorption of serum proteins. Simultaneously, numerical models were developed to simulate the mass transport of chemical species to and from the BG surface under normal gravity and simulated microgravity conditions. The numerical results showed an excellent agreement with the experimental data at both testing conditions.

Radin, Shula; Ducheyne, P.; Ayyaswamy, P. S.

2003-01-01

303

Integrative Proteomics and Tissue Microarray Profiling Indicate the Association between Overexpressed Serum Proteins and Non-Small Cell Lung Cancer  

PubMed Central

Lung cancer is the leading cause of cancer deaths worldwide. Clinically, the treatment of non-small cell lung cancer (NSCLC) can be improved by the early detection and risk screening among population. To meet this need, here we describe the application of extensive peptide level fractionation coupled with label free quantitative proteomics for the discovery of potential serum biomarkers for lung cancer, and the usage of Tissue microarray analysis (TMA) and Multiple reaction monitoring (MRM) assays for the following up validations in the verification phase. Using these state-of-art, currently available clinical proteomic approaches, in the discovery phase we confidently identified 647 serum proteins, and 101 proteins showed a statistically significant association with NSCLC in our 18 discovery samples. This serum proteomic dataset allowed us to discern the differential patterns and abnormal biological processes in the lung cancer blood. Of these proteins, Alpha-1B-glycoprotein (A1BG) and Leucine-rich alpha-2-glycoprotein (LRG1), two plasma glycoproteins with previously unknown function were selected as examples for which TMA and MRM verification were performed in a large sample set consisting about 100 patients. We revealed that A1BG and LRG1 were overexpressed in both the blood level and tumor sections, which can be referred to separate lung cancer patients from healthy cases. PMID:23284758

Hu, Haichuan; Wang, Rui; Sun, Yihua; Zeng, Rong; Chen, Haiquan

2012-01-01

304

Jun is phosphorylated by several protein kinases at the same sites that are modified in serum-stimulated fibroblasts.  

PubMed Central

c-jun is a member of the family of immediate-early genes whose expression is induced by factors such as serum stimulation, phorbol ester, and differentiation signals. Here we show that increased Jun synthesis after serum stimulation is accompanied by a concomitant increase in phosphorylation. Several serine-threonine kinases were evaluated for their ability to phosphorylate Jun in vitro. p34cdc2, protein kinase C, casein kinase II, and pp44mapk phosphorylated Jun efficiently, whereas cyclic AMP-dependent protein kinase and glycogen synthase kinase III did not. The sites phosphorylated by p34cdc2 were similar to those phosphorylated in vivo after serum induction. The major sites of phosphorylation were mapped to serines 63, 73, and 246. Phosphorylation of full-length Jun with several kinases did not affect the DNA-binding activity of Jun homodimers or Fos-Jun heterodimers. Comparison of the DNA binding and in vitro transcription properties of wild-type and mutated proteins containing either alanine or aspartic acid residues in place of Ser-63, -73, and -246 revealed only minor differences among homodimeric complexes and no differences among Fos-Jun heterodimers. Thus, phosphorylation of Jun did not produce a significant change in dimerization, DNA-binding, or in vitro transcription activity. The regulatory role of phosphorylation in the modulation of Jun function is likely to be considerably more complex than previously suggested. Images PMID:1328860

Baker, S J; Kerppola, T K; Luk, D; Vandenberg, M T; Marshak, D R; Curran, T; Abate, C

1992-01-01

305

Hierarchical role of fetuin-A and acidic serum proteins in the formation and stabilization of calcium phosphate particles.  

PubMed

The serum protein fetuin-A is a potent systemic inhibitor of soft tissue calcification. Fetuin-A is highly effective in the formation and stabilization of protein-mineral colloids, referred to as calciprotein particles (CPPs). These particles ripen in vitro in a two-step process, indicated by a morphological conversion from spheres to larger prolate ellipsoids. Using a combined light scattering and electron microscopic imaging approach we determined that the second-stage particles resulted from a highly anisotropic outgrowth of the first-stage particles. Electron microscopy of ascites fluid from a patient with calcifying peritonitis revealed particles reminiscent of secondary CPPs. Thus, CPPs form in the body and undergo the two-step ripening at least in pathological conditions. Unlike in vitro generated CPPs, ascites-derived CPPs contained little fetuin-A but large amounts of albumin. This prompted us to study the role of fetuin-A combined with other serum proteins in CPP formation. Fetuin-A was indispensable for primary CPP formation. Albumin and acidic proteins in general greatly enhanced the fetuin-A triggered formation of secondary CPPs and, thus, substituted substantial amounts of fetuin-A without loss of inhibition of calcium phosphate precipitation. Thus, direct mineral deposition from solute in the body is unlikely even at low fetuin-A serum levels as long as sufficient bulk acidic protein is available. Collectively fetuin-A and other acidic bulk plasma proteins may be considered as mineral chaperones mediating the stabilization, safe transport, and clearance in the body of calcium and phosphate as colloidal complexes, thus, preventing ectopic calcification. PMID:18364352

Heiss, Alexander; Eckert, Thomas; Aretz, Anke; Richtering, Walter; van Dorp, Wim; Schäfer, Cora; Jahnen-Dechent, Willi

2008-05-23

306

Implant debris particle size affects serum protein adsorption which may contribute to particle size-based bioreactivity differences  

PubMed Central

Biologic reactivity to orthopedic implant debris mediates long-term clinical performance of total joint arthroplasty implants. However, why some facets of implant debris are more pro-inflammatory remains controversial such as particle size, shape, base material etc. This precludes accurate prediction and optimal design of modern total joint replacements. We hypothesized that debris particle size can influence adsorbed protein film composition and affect subsequent bioreactivity. We measured size-dependent protein film-adsorption, and adsorbed protein film-dependent cytokine release using equal surface areas of different sized cobalt-chromium-alloy (CoCr-alloy) particle and in vitro challenge of human macrophages (THP-1 and human primary). Smaller 5?m vs 70?m sized particles preferentially adsorbed more serum protein in general (p<0.03), where higher molecular weight serum proteins consistent with IgG were identified. Additionally, 5?m CoCr-alloy particles pre-coated with different protein biofilms (IgG vs albumin) resulted in differential cytokine expression where albumin-coated particles induced more TNF-? and IgG-coated particles induced more IL-1? release from human monocyte/macrophages. In these preliminary in vitro studies we demonstrated the capability of equal surface areas of different particle sizes to influence adsorbed protein composition and that adsorbed protein differences on identical particles can translate into complex differences in bioreactivity. Together this suggests adsorbed protein differences on different sized particles of the same material may be a contributing mechanism by which different sized particles induce differences in reactivity. PMID:24941408

Reddy, Anand; Caicedo, Marco; Samelko, Lauryn; Jacobs, Joshua J; Hallab, Nadim James

2014-01-01

307

Zeptomole Detection of C-Reactive Protein in Serum by a Nanoparticle Amplified Surface Plasmon Resonance Imaging Aptasensor  

NASA Astrophysics Data System (ADS)

Diagnostic biomarkers (i.e. proteins) are often in low abundance in bodily fluids presenting many challenges for their detection. In order to extend the application of SPRi systems in detecting biomarkers at ultralow levels, we combine the advantage of aptamer technology with nanomaterials and microwave-assisted surface functionalization. By implementing a sandwich assay through the introduction of aptamer-modified quantum dots (QDs), it was possible to measure 7 zeptomole (at 5 fg/mL) of C-reactive protein (CRP) selectively in spiked human serum. It is expected that the proposed platform will provide new direction in designing ultrasensitive SPRi biosensors with multiplexing capabilities.

Vance, Stephen A.; Sandros, Marinella G.

2014-05-01

308

Zeptomole Detection of C-Reactive Protein in Serum by a Nanoparticle Amplified Surface Plasmon Resonance Imaging Aptasensor  

PubMed Central

Diagnostic biomarkers (i.e. proteins) are often in low abundance in bodily fluids presenting many challenges for their detection. In order to extend the application of SPRi systems in detecting biomarkers at ultralow levels, we combine the advantage of aptamer technology with nanomaterials and microwave-assisted surface functionalization. By implementing a sandwich assay through the introduction of aptamer-modified quantum dots (QDs), it was possible to measure 7 zeptomole (at 5?fg/mL) of C-reactive protein (CRP) selectively in spiked human serum. It is expected that the proposed platform will provide new direction in designing ultrasensitive SPRi biosensors with multiplexing capabilities. PMID:24875139

Vance, Stephen A.; Sandros, Marinella G.

2014-01-01

309

Sandwich enzyme-linked immunosorbent assay for quantitative measurement of serum amyloid A protein in horses.  

PubMed

To measure the concentration of serum amyloid A (sAA) protein in horses, a sensitive and highly reproducible sandwich (ELISA) was established, using affinity purified SAA antibody. Results of the ELISA were found to have a high correlation (r = 0.95) with those of the single radial immunodiffusion test. Equine SAA concentration was measured by use of this ELISA. In clinically normal horses, the concentration of SAA was high immediately after birth to 2 weeks of age. After that, SAA concentration had periodic fluctuations in the range of approximately 1.0 to 30 micrograms/ml. Mean (+/- SD)) concentrations of SAA in foals (< or = 12 months old) and adult horses (> or = 18 months old) were 21.23 +/- 12.20 and 14.93 +/- 9.07 micrograms/ml, respectively. In mares during the perinatal period, SAA concentration remained stable within the reference range before parturition. It increased quickly after delivery, and reached a peak value of 101.29 +/- 98.82 micrograms/ml on postpartum day 3, then began to decrease, at postpartum week 2, to the reference range by the end of postpartum month 1. In horses with experimentally induced inflammation, SAA concentration increased quickly and reached approximately four- to 40-fold increase over the pretreatment value on day 1 and remained high on days 2 to 6 after treatment. It then returned to the baseline value by 2 to 4 weeks in association with disappearance of local signs of inflammation. The SAA concentration was high in most horses with clinical signs of inflammation. It was concluded from these data that this ELISA is sensitive and reliable for measuring SAA in horses. PMID:8928944

Satoh, M; Fujinaga, T; Okumura, M; Hagio, M

1995-10-01

310

Association Between Dietary Pattern and Serum C-Reactive Protein in Japanese Men and Women  

PubMed Central

Background Dietary pattern may influence the risks of cardiovascular disease, atherosclerosis, type 2 diabetes, and metabolic syndrome through its effects on inflammation. We evaluated the association between dietary pattern and serum high-sensitivity C-reactive protein (hs-CRP) in a Japanese population. Methods In this cross-sectional analysis, we used baseline data from 3905 men and 5640 women (age 40–69 years) who participated in a population-based cohort study between November 2005 and December 2007. Participants with possible inflammation-related diseases, current analgesic use, high hs-CRP levels (?3000 ng/mL) or extreme dietary energy intake were excluded. We used 46 items from a validated short food frequency questionnaire and examined major dietary patterns by factor analysis. Results We identified 5 dietary patterns: healthy (high in vegetables and fruit), Western (high in meat and fried foods), seafood (high in shellfish, squid, fish, etc.), bread (high in bread and low in rice), and dessert (high in confections and fruit). After adjustment for age, alcohol use, smoking, physical activity, and body mass index, hs-CRP levels in men were inversely associated with the healthy, bread, and dessert patterns (P-trend: 0.01, 0.06, and <0.01, respectively) and positively associated with the seafood pattern (P-trend = 0.02). In women, hs-CRP levels were inversely associated with the healthy pattern (P-trend = 0.06) and positively associated with the Western pattern (P-trend = 0.06). Conclusions The healthy dietary pattern may be associated with suppressed inflammation in Japanese men and women, independently of body mass index and other factors. The sex-specific associations of hs-CRP with other dietary patterns (eg, the seafood pattern) require further study. PMID:21325731

Nanri, Hinako; Nakamura, Kazuyo; Hara, Megumi; Higaki, Yasuki; Imaizumi, Takeshi; Taguchi, Naoto; Sakamoto, Tatsuhiko; Horita, Mikako; Shinchi, Koichi; Tanaka, Keitaro

2011-01-01

311

Lowering of Serum Cholesteryl Ester Transfer Protein—But Not Lecithin:Cholesterol Acyltransferase—Activity Levels by Hypocholesterolemic Drugs in the Rabbit  

Microsoft Academic Search

Summary. Cholesteryl ester transfer protein (CETP) and lecithin:cholesterol acyltransferase (LCAT) are important factors in the regulation of serum lipoprotein metabolism. Rabbits were fed hypocholesterolemic drugs to investigate the effect on serum CETP and LCAT activity levels. The activities were assayed using exogenous substrate assays and are an estimate of CETP and LCAT mass. Groups of eight rabbits were fed a

G. W. Meijer; J. E. M. Groener; A. C. Beynen; A. Van Tol

1998-01-01

312

Serum C-reactive protein predicts poor prognosis in patients with locoregionally advanced nasopharyngeal carcinoma treated with chemoradiotherapy  

PubMed Central

Background We aimed to evaluate the association of serum C-reactive protein (crp) with prognosis in patients with locoregionally advanced nasopharyngeal carcinoma treated with chemoradiotherapy. Methods We retrospectively reviewed 79 patients with locoregionally advanced nasopharyngeal carcinoma (cT3–4N0–3M0) treated with chemoradiotherapy. Chemoradiotherapy consisted of external-beam radiotherapy to the nasopharynx (70–80 Gy), the lymph node–positive area (60–70 Gy), and the lymph node–negative area (50–60 Gy) combined with 3 cycles of various platinum-based regimens delivered at 3-week intervals. Elevated crp was defined as more than 8 mg/L. The survival rate was calculated using the Kaplan–Meier method, and univariate and multivariate analyses (Cox proportional hazards model) were used to identify factors significantly associated with prognosis. Results During the median follow-up of 3.9 years (range: 1–5.5 years), 23 patients died from nasopharyngeal cancer. The 5-year cancer-specific survival (css) rate was 62.90%. Before chemoradiotherapy, 18 patients had high serum crp; the css rate in that subgroup was significantly worse than the rate in the remaining patients (p = 0.0002). Multivariate analysis showed that crp was an independent prognostic indicator of css, with a hazard ratio of 3.04 (95% confidence interval: 1.22 to 7.55; p = 0.017). Among the 18 patients with elevated serum crp, 9 achieved normal serum crp after chemoradiotherapy, of whom 5 remained living with no evidence of recurrence or metastasis during follow-up. By contrast, the remaining 9 patients in whom serum crp did not normalize after chemoradiotherapy died within 4.2 years. Conclusions Elevated serum crp before treatment predicts poor prognosis in patients with locoregionally advanced nasopharyngeal carcinoma treated with chemoradiotherapy.

Zeng, Y.C.; Wu, R.; Xiao, Y.P.; Chi, F.; Xue, M.; Zhang, Z.Y.; Xing, R.; Zhong, W.Z.; Wang, S.L.; Tian, X.; Chen, W.; Chen, J.J.; Wu, L.N.

2015-01-01

313

Combined Inflammatory and Metabolic Defects Reflected by Reduced Serum Protein Levels in Patients with Buruli Ulcer Disease  

PubMed Central

Buruli ulcer is a skin disease caused by Mycobacterium ulcerans that is spreading in tropical countries, with major public health and economic implications in West Africa. Multi-analyte profiling of serum proteins in patients and endemic controls revealed that Buruli ulcer disease down-regulates the circulating levels of a large array of inflammatory mediators, without impacting on the leukocyte composition of peripheral blood. Notably, several proteins contributing to acute phase reaction, lipid metabolism, coagulation and tissue remodelling were also impacted. Their down-regulation was selective and persisted after the elimination of bacteria with antibiotic therapy. It involved proteins with various functions and origins, suggesting that M. ulcerans infection causes global and chronic defects in the host's protein metabolism. Accordingly, patients had reduced levels of total serum proteins and blood urea, in the absence of signs of malnutrition, or functional failure of liver or kidney. Interestingly, slow healers had deeper metabolic and coagulation defects at the start of antibiotic therapy. In addition to providing novel insight into Buruli ulcer pathogenesis, our study therefore identifies a unique proteomic signature for this disease. PMID:24722524

Landier, Jordi; Oldenburg, Reid; Frimpong, Michael; Wansbrough-Jones, Mark; Abass, Kabiru; Thompson, William; Forson, Mark; Fontanet, Arnaud; Niang, Fatoumata; Demangel, Caroline

2014-01-01

314

EFFECT OF WHOLE-BODY X-IRRADIATION ON SERUM PROTEIN OF RABBITS (EFFECT OF WHOLE BODY X-IRRADIATION AT ONE TIME)  

Microsoft Academic Search

The various fractions of the serum proteins of xirradiated rabbits were ; measured by the paper-electrophoretic method. After 400 r whole-body ; irradiation, total protein and albumin were decreased, while alpha â-, ; alpha â and gamma globulins were increased. After 1,000 r whole-body ; irradiation, total protein, albumin, and gamma -globulin were decreased with an ; increase of alpha

Yoshino

1960-01-01

315

Associations of leisure time physical activity, self-rated physical fitness, and estimated aerobic fitness with serum C-reactive protein among 3803 adults  

Microsoft Academic Search

ObjectiveSerum C-reactive protein (CRP), a marker of systemic inflammation, is a risk factor for cardiovascular disease. Obesity and physical activity are associated with CRP, though population studies are sparse.

Borodulin Katja; Tiina Laatikainen; Veikko Salomaa; Pekka Jousilahti

2006-01-01

316

Larval serum proteins of the gypsy moth, Lymantria dispar: Allometric changes during development suggest several functions for arylphorin and lipophorin  

SciTech Connect

Storage proteins are the major nutritive intermediates in insects and although the serum storage proteins are relatively well studied, definitive roles for many of them have yet to be established. To further characterize their roles in development and to establish quantitative baselines for future studies, two serum proteins, arylphorin (Ap) and lipophorin (Lp), of the gypsy moth, Lymantria dispar, were studied. Ap and Lp, isolated from larval hemolymph, were partially characterized biochemically and immunologically. Hemolymph concentrations throughout larval development were determined using quantitative immunoelectrophoresis and absolute hemolymph amounts of protein were determined by measuring hemolymph volume. Cyclic fluctuations in hemolymph concentrations of Ap in particular correlated with each molting cycle and an increase in Lp levels just prior to pupation suggest a metamorphic change in the role or demand for the protein. Sexual dimorphism in protein concentrations are explained in part by the sexual dimorphism in the number of larval instars. In fact, an additional instar of Ap accumulation in the female gypsy moth is suggested to compensate for the lack of a female-specific storage protein in this species. The last two days of each instar were found to be the optimum time to sample protein concentration with minimum variance. Allometric relationships among Ap accumulation, Lp accumulation and weight gain were uncovered. Ap labelled with ({sup 14}C)-N-ethylmaleimide was shown to be incorporated into newly synthesized cuticle and setae during a larval-larval molt. The antiserum developed against L. dispar Ap was used to identify the Ap of Trichoplusia in and study Ap titers in parasitized T. in larvae. The antiserum was also used to determine the immunological relatedness of 5 species of Lepidoptera.

Karpells, S.T.

1989-01-01

317

Serum immunoglobulin A and antibody to M-associated protein in patients with acute glomerulonephritis or rheumatic fever.  

PubMed Central

Serum immunoglobulin A (IgA) was markedly increased in 80% of 50 patients with acute rheumatic fever in Trinidad in contrast to 20% of 63 patients with acute glomerulonephritis, whereas serum IgG was increased in nearly all of both groups. Since total antibody to M-associated protein (MAP) (assessed by complement fixation) is the only antibody as yet consistently found to be of higher titer in patients with acute rheumatic fever than in patients with acute glomerulonephritis in Trinidad, it was measured and the titers were related to serum levels of IgA. The titers of total antibody to MAP were greater than or equal to 40 in 58% of the rheumatic fever patients, 43% of the nephritis patients, and 8% of well school-children. However, its presence in rheumatic fever patients did not correlate directly with amounts of serum IgA present (r = 0.0507). Moreover, titers of total antibody to MAP related equally well to enzyme-linked immunosorbent assay indicated amounts of IgG antibody (r = 0.3939) and IgA antibody (r = 0.3054) to MAP in rheumatic fever patients but not in nephritis patients (r = 0.0301 for IgA antibody and 0.6909 for IgG antibody to MAP), whereas they related best to IgM antibody to MAP in the school-children (r = 0.4204). PMID:7107003

Potter, E V; Shaughnessy, M A; Poon-King, T; Earle, D P

1982-01-01

318

Serum response factor and protein-mediated DNA bending contribute to transcription of the dystrophin muscle-specific promoter.  

PubMed Central

The minimal muscle-specific dystrophin promoter contains the consensus sequence CC(A/T)6GG, or the CArG element, which can be found in serum-inducible or muscle-specific promoters. The serum response factor (SRF), which mediates the transcriptional activation of the c-fos gene in response to serum stimulation, can bind to different CArG box elements, suggesting that it could be involved in muscle-constitutive transcription. Here we show that SRF binds to the dystrophin promoter and regulates its muscle-specific transcription. In transient transfections, an altered-binding-specificity SRF mutant restores the muscle-constitutive transcription of a dystrophin promoter with a mutation in its CArG box element. The muscle-constitutive transcription of the dystrophin promoter also requires the sequence GAAACC immediately downstream of the CArG box. This sequence is recognized by a novel DNA bending factor which was named dystrophin promoter-bending factor (DPBF). Mutations of the CArG flanking sequence abolish both DPBF binding and the promoter activity in muscle cells. Its replacement with a p62/ternary complex factor binding site changes the promoter specificity from muscle constitutive to serum responsive. These results show that, on the dystrophin promoter, the transcriptional activation induced by SRF requires the DNA bending induced by DPBF. The bending, next to the CArG box, could promote interactions between SRF and other proteins in the transcriptional complex. PMID:9032300

Galvagni, F; Lestingi, M; Cartocci, E; Oliviero, S

1997-01-01

319

Isolation of Protein-Associated Circular DNA from Healthy Cattle Serum  

PubMed Central

Three replication-competent single-stranded DNA molecules sharing nucleotide similarity to transmissible spongiform encephalopathy (TSE)-associated isolate Sphinx 2.36 were isolated from healthy bovine serum. PMID:25169856

Funk, Mathis; Gunst, Karin; Lucansky, Vincent; Müller, Hermann; zur Hausen, Harald

2014-01-01

320

Isolation of protein-associated circular DNA from healthy cattle serum.  

PubMed

Three replication-competent single-stranded DNA molecules sharing nucleotide similarity to transmissible spongiform encephalopathy (TSE)-associated isolate Sphinx 2.36 were isolated from healthy bovine serum. PMID:25169856

Funk, Mathis; Gunst, Karin; Lucansky, Vincent; Müller, Hermann; Zur Hausen, Harald; de Villiers, Ethel-Michele

2014-01-01

321

Quantification of Selected Serum Proteins of Milk by Immunological Procedures1  

Microsoft Academic Search

Evaluation was of these immunological procedures for the quantification of the bovine serum albumin and\\/3-1actoglobulin content of milk: Oudin's single diffusion, Mancini's radial immunodiffusion, and Laurell's electroimmunodiffusion tech- niques. All three methods were repro- ducible and did not require preliminary treatment of the milk. The bovine serum albumin content (mg\\/ml) of 25 different herd and bulk milk samples ranged from

Maria Babajimopoulos; E. M. Mikolajcik

1977-01-01

322

Comparison of the Serum Protein Fractions of the Newly Hatched Chick with those of Adult Birds using Starch-gel Electrophoresis  

Microsoft Academic Search

IT has been established by several investigators that serum protein components of chick embryo undergo qualitative as well as quantitative changes throughout the developmental period until the time of hatching1-4. The serum protein composition of the newly hatched chick, however, resembles that of the adult bird, as revealed by moving-boundary and paper electro-phoresis. The technique of starch-gel electrophoresis suggested by

A. Amin

1961-01-01

323

A novel V(IV)O-pyrimidinone complex: synthesis, solution speciation and human serum protein binding.  

PubMed

The pyrimidinones mhcpe, 2-methyl-3H-5-hydroxy-6-carboxy-4-pyrimidinone ethyl ester (mhcpe, 1), 2,3-dimethyl-5-benzyloxy-6-carboxy-4-pyrimidinone ethyl ester (dbcpe, 2) and N-methyl-2,3-dimethyl-5-hydroxy-6-carboxyamido-4-pyrimidinone (N-MeHOPY, 3), are synthesized and their structures determined by single crystal X-ray diffraction. The acid-base properties of 1 are studied by potentiometric and spectrophotometric methods, the pK(a) values being 1.14 and 6.35. DFT calculations were carried out to determine the most stable structure for each of the H2L(+), HL and L(-) forms (HL = mhcpe) and assign the groups involved in the protonation-deprotonation processes. The mhcpe(-) ligand forms stable complexes with V(IV)O(2+) in the pH range 2 to 10, and potentiometry, EPR and UV-Vis techniques are used to identify and characterize the V(IV)O-mhcpe species formed. The results are consistent with the formation of V(IV)O, (V(IV)O)L, (V(IV)O)L2, (V(IV)O)2L2H(-2), (V(IV)O)L2H(-1), (V(IV)O)2L2H(-3), (V(IV)O)LH(-2) species and V(IV)O-hydrolysis products. Calculations indicate that the global binding ability of mhcpe towards V(IV)O(2+) is similar to that of maltol (Hmaltol = 3-hydroxy-2-methyl-4H-pyran-4-one) and lower than that of 1,2-dimethyl-3-hydroxy-4-pyridinone (Hdhp). The interaction of V(IV)O-complexes with human plasma proteins (transferrin and albumin) is studied by circular dichroism (CD), EPR and (51)V NMR spectroscopy. V(IV)O-mhcpe-protein ternary complexes are formed in both cases. The binding of V(IV)O(2+) to transferrin (hTF) in the presence of mhcpe involves mainly (V(IV)O)1(hTF)(mhcpe)1, (V(IV)O)2(hTF)(mhcpe)1 and (V(IV)O)2(hTF)(mhcpe)2 species, bound at the Fe(III) binding sites, and the corresponding conditional formation constants are determined. Under the conditions expected to prevail in human blood serum, CD data indicate that the V(IV)O-mhcpe complexes mainly bind to hTF; the formation of V(IV)O-hTF-mhcpe complexes occurs in the presence of Fe(III) as well, distinct EPR signals being clearly obtained for Fe(III)-hTF and to V(IV)O-hTF-mhcpe species. Thus this study indicates that transferrin plays the major role in the transport of V(IV)O-mhcpe complexes under blood plasma conditions in the form of ternary V(IV)-ligand-protein complexes. PMID:23677414

Gonçalves, Gisela; Tomaz, Isabel; Correia, Isabel; Veiros, Luís F; Castro, M Margarida C A; Avecilla, Fernando; Palacio, Lorena; Maestro, Miguel; Kiss, Tamás; Jakusch, Tamás; Garcia, M Helena V; Pessoa, João Costa

2013-09-01

324

Phosphorylation of Heat Shock Protein 27 is Increased by Cast Immobilization and by Serum-free Starvation in Skeletal Muscles.  

PubMed

[Purpose] Cast immobilization- and cell starvation-induced loss of muscle mass are closely associated with a dramatic reduction in the structural muscle proteins. Heat shock proteins are molecular chaperones that are constitutively expressed in several eukaryotic cells and have been shown to protect against various stressors. However, the changes in the phosphorylation of atrophy-related heat shock protein 27 (HSP27) are still poorly understood in skeletal muscles. In this study, we examine whether or not phosphorylation of HSP27 is changed in the skeletal muscles after cast immobilization and serum-free starvation with low glucose in a time-dependent manner. [Methods] We undertook a HSP27 expression and high-resolution differential proteomic analysis in skeletal muscles. Furthermore, we used western blotting to examine protein expression and phosphorylation of HSP27 in atrophied gastrocnemius muscle strips and L6 myoblasts. [Results] Cast immobilization and starvation significantly upregulated the phosphorylation of HSP27 in a time-dependent manner, respectively. [Conclusion] Our results suggest that cast immobilization- and serum-free starvation-induced atrophy may be in part related to changes in the phosphorylation of HSP27 in rat skeletal muscles. PMID:25540511

Kim, Mee-Young; Lee, Jeong-Uk; Kim, Ju-Hyun; Lee, Lim-Kyu; Park, Byoung-Sun; Yang, Seung-Min; Jeon, Hye-Joo; Lee, Won-Deok; Noh, Ji-Woong; Kwak, Taek-Yong; Jang, Sung-Ho; Lee, Tae-Hyun; Kim, Ju-Young; Kim, Bokyung; Kim, Junghwan

2014-12-01

325

Phosphorylation of Heat Shock Protein 27 is Increased by Cast Immobilization and by Serum-free Starvation in Skeletal Muscles  

PubMed Central

[Purpose] Cast immobilization- and cell starvation-induced loss of muscle mass are closely associated with a dramatic reduction in the structural muscle proteins. Heat shock proteins are molecular chaperones that are constitutively expressed in several eukaryotic cells and have been shown to protect against various stressors. However, the changes in the phosphorylation of atrophy-related heat shock protein 27 (HSP27) are still poorly understood in skeletal muscles. In this study, we examine whether or not phosphorylation of HSP27 is changed in the skeletal muscles after cast immobilization and serum-free starvation with low glucose in a time-dependent manner. [Methods] We undertook a HSP27 expression and high-resolution differential proteomic analysis in skeletal muscles. Furthermore, we used western blotting to examine protein expression and phosphorylation of HSP27 in atrophied gastrocnemius muscle strips and L6 myoblasts. [Results] Cast immobilization and starvation significantly upregulated the phosphorylation of HSP27 in a time-dependent manner, respectively. [Conclusion] Our results suggest that cast immobilization- and serum-free starvation-induced atrophy may be in part related to changes in the phosphorylation of HSP27 in rat skeletal muscles.

Kim, Mee-Young; Lee, Jeong-Uk; Kim, Ju-Hyun; Lee, Lim-Kyu; Park, Byoung-Sun; Yang, Seung-Min; Jeon, Hye-Joo; Lee, Won-Deok; Noh, Ji-Woong; Kwak, Taek-Yong; Jang, Sung-Ho; Lee, Tae-Hyun; Kim, Ju-Young; Kim, Bokyung; Kim, Junghwan

2014-01-01

326

Protein signature-based estimation of metagenomic abundances including all domains of life and viruses  

PubMed Central

Motivation: Metagenome analysis requires tools that can estimate the taxonomic abundances in anonymous sequence data over the whole range of biological entities. Because there is usually no prior knowledge about the data composition, not only all domains of life but also viruses have to be included in taxonomic profiling. Such a full-range approach, however, is difficult to realize owing to the limited coverage of available reference data. In particular, archaea and viruses are generally not well represented by current genome databases. Results: We introduce a novel approach to taxonomic profiling of metagenomes that is based on mixture model analysis of protein signatures. Our results on simulated and real data reveal the difficulties of the existing methods when measuring achaeal or viral abundances and show the overall good profiling performance of the protein-based mixture model. As an application example, we provide a large-scale analysis of data from the Human Microbiome Project. This demonstrates the utility of our method as a first instance profiling tool for a fast estimate of the community structure. Availability: http://gobics.de/TaxyPro. Contact: pmeinic@gwdg.de Supplementary information: Supplementary Material is available at Bioinformatics online. PMID:23418187

Klingenberg, Heiner; Aßhauer, Kathrin Petra; Lingner, Thomas; Meinicke, Peter

2013-01-01

327

A novel multiplex-protein array for serum diagnostics of colon cancer: a case–control study  

PubMed Central

Background More than 1.2 million new cases of colorectal cancer are reported each year worldwide. Despite actual screening programs, about 50% of the patients are diagnosed at advanced tumor stages presenting poor prognosis. Innovative screening tools could aid the detection at early stages and allow curative treatment interventions. Methods A nine target multiplex serum protein biochip was generated and evaluated using a training- and validation-set of 317 highly standardized, liquid nitrogen preserved serum samples comprising controls, adenomas, and colon cancers. Results Serum levels of CEA, IL-8, VEGF, S100A11, MCSF, C3adesArg, CD26, and CRP showed significant differences between cases and controls. The largest areas under the receiver operating characteristics curve were observed for CEA, IL-8, and CRP. At threshold levels yielding 90% specificity, sensitivities for CEA, IL-8 and CRP were 26%, 22%, and 17%, respectively. The most promising marker combinations were CEA?+?IL-8 reaching 37% sensitivity at 83% specificity and CEA?+?CRP with 35% sensitivity at 81% specificity. In an independent validation set CEA?+?IL-8 reached 47% sensitivity at 86% specificity while CEA?+?CRP obtained 39% sensitivity at 86% specificity. Early carcinomas were detected with 33% sensitivity for CEA?+?IL-8 and 28% for CEA?+?CRP. Conclusions Apart from CEA, IL-8, and CRP, the screening value of additional blood markers and the potential advantage of combining serum biochip testing with fecal occult blood testing needs to be studied. Multiplex biochip array technology utilizing serum samples offers an innovative approach to colorectal cancer screening. PMID:22954206

2012-01-01

328

Dietary total antioxidant capacity from different assays in relation to serum C-reactive protein among young Japanese women  

PubMed Central

Background The association between dietary total antioxidant capacity (TAC) from different assays and serum C-reactive protein (CRP) has not been assessed in non-Western populations. We examined the association between dietary TAC and serum CRP concentration in young Japanese women using different four TAC assays. Methods The subjects were 443 young Japanese women aged 18–22?years. Dietary TAC was assessed with a self-administered diet history questionnaire and the TAC value of each food using the following four assays: ferric reducing ability of plasma (FRAP); oxygen radical absorbance capacity (ORAC); Trolox equivalent antioxidant capacity (TEAC); and total radical-trapping antioxidant parameter (TRAP). Serum CRP concentrations were measured by highly sensitive nephelometry. Results The major contributor to dietary TAC was green, barley, and oolong tea (FRAP: 53%, ORAC: 45%, TEAC: 36%, and TRAP: 44%). The prevalence of elevated CRP concentrations (? 1?mg/L) was 5.6%. TAC from FRAP was inversely associated with serum CRP concentrations (adjusted odds ratio [OR] for elevated CRP concentration in high [compared with low] dietary TAC group: 0.39 [95% confidence interval (CI): 0.16-0.98]; P?=?0.04). TAC from ORAC was inversely associated with CRP, although the association was not significant (OR: 0.48 [95% CI: 0.20-1.14]; P?=?0.10). TAC from TEAC was inversely associated with CRP (OR: 0.32 [95% CI: 0.12-0.82]; P?=?0.02), as was TAC from TRAP (OR: 0.31 [95% CI: 0.12-0.81]; P?=?0.02). Conclusions Dietary TAC was inversely associated with serum CRP concentration in young Japanese women regardless of assay. Further studies are needed in other populations to confirm these results. PMID:23110638

2012-01-01

329

Uncoupling protein 2 prevents neuronal death including that occurring during seizures: a mechanism for preconditioning.  

PubMed

The mitochondrial uncoupling protein (UCP2) is expressed in selected regions of the brain. Here we demonstrate that up-regulation of UCP2 is part of a neuroprotective set of responses to various cellular stresses in vitro and in vivo. PC12 cells, when transfected with UCP2, were protected against free radical-induced cell death. Seizure activity was associated with elevated UCP2 levels and mitochondrial uncoupling activity. In transgenic mice that expressed UCP2 constitutively in the hippocampus before seizure induction, a robust reduction in cell death was seen. Because UCP2 increased mitochondrial number and ATP levels with a parallel decrease in free radical-induced damage, it is reasonable to suggest that mitochondrial UCPs precondition neurons by dissociating cellular energy production from that of free radicals to withstand the harmful effects of cellular stress occurring in a variety of neurodegenerative disorders, including epilepsy. PMID:12960023

Diano, Sabrina; Matthews, Russell T; Patrylo, Peter; Yang, Lichuan; Beal, M Flint; Barnstable, Colin J; Horvath, Tamas L

2003-11-01

330

Rapid development of sensitive, high-throughput, quantitative and highly selective mass spectrometric targeted immunoassays for clinically important proteins in human plasma and serum  

PubMed Central

Objectives The aim of this study was to develop high-throughput, quantitative and highly selective mass spectrometric, targeted immunoassays for clinically important proteins in human plasma or serum. Design and methods The described method coupled mass spectrometric immunoassay (MSIA), a previously developed technique for immunoenrichment on a monolithic microcolumn activated with an anti-protein antibody and fixed in a pipette tip, to selected reaction monitoring (SRM) detection and accurate quantification of targeted peptides, including clinically relevant sequence or truncated variants. Results In this report, we demonstrate the rapid development of MSIA-SRM assays for sixteen different target proteins spanning seven different clinically important areas (including neurological, Alzheimer's, cardiovascular, endocrine function, cancer and other diseases) and ranging in concentration from pg/mL to mg/mL. The reported MSIA-SRM assays demonstrated high sensitivity (within published clinical ranges), precision, robustness and high-throughput as well as specific detection of clinically relevant isoforms for many of the target proteins. Most of the assays were tested with bona-fide clinical samples. In addition, positive correlations, (R2 0.67–0.87, depending on the target peptide), were demonstrated for MSIA-SRM assay data with clinical analyzer measurements of parathyroid hormone (PTH) and insulin growth factor 1 (IGF1) in clinical sample cohorts. Conclusions We have presented a practical and scalable method for rapid development and deployment of MS-based SRM assays for clinically relevant proteins and measured levels of the target analytes in bona fide clinical samples. The method permits the specific quantification of individual protein isoforms and addresses the difficult problem of protein heterogeneity in clinical proteomics applications. PMID:23313081

Krastins, Bryan; Prakash, Amol; Sarracino, David A.; Nedelkov, Dobrin; Niederkofler, Eric E.; Kiernan, Urban A.; Nelson, Randall; Vogelsang, Maryann S.; Vadali, Gouri; Garces, Alejandra; Sutton, Jennifer N.; Peterman, Scott; Byram, Gregory; Darbouret, Bruno; Pérusse, Joëlle R.; Seidah, Nabil G.; Coulombe, Benoit; Gobom, Johan; Portelius, Erik; Pannee, Josef; Blennow, Kaj; Kulasingam, Vathany; Couchman, Lewis; Moniz, Caje; Lopez, Mary F.

2013-01-01

331

Interdialytic weight gain, systolic blood pressure, serum albumin, and C-reactive protein levels change in chronic dialysis patients prior to death.  

PubMed

Reports from a United States cohort of chronic hemodialysis patients suggested that weight loss, a decline in pre-dialysis systolic blood pressure, and decreased serum albumin may precede death. However, no comparative studies have been reported in such patients from other countries. Here we analyzed dynamic changes in these parameters in hemodialysis patients and included 3593 individuals from 5 Asian countries; 35,146 from 18 European countries; 8649 from Argentina; and 4742 from the United States. In surviving prevalent patients, these variables appeared to have notably different dynamics than in patients who died. While in all populations the interdialytic weight gain, systolic blood pressure, and serum albumin levels were stable in surviving patients, these indicators declined starting more than a year ahead in those who died with the dynamics similar irrespective of gender and geographic region. In European patients, C-reactive protein levels were available on a routine basis and indicated that levels of this acute-phase protein were low and stable in surviving patients but rose sharply before death. Thus, relevant fundamental biological processes start many months before death in the majority of chronic hemodialysis patients. Longitudinal monitoring of these dynamics may help to identify patients at risk and aid the development of an alert system to initiate timely interventions to improve outcomes. PMID:23515055

Usvyat, Len A; Barth, Claudia; Bayh, Inga; Etter, Michael; von Gersdorff, Gero D; Grassmann, Aileen; Guinsburg, Adrian M; Lam, Maggie; Marcelli, Daniele; Marelli, Cristina; Scatizzi, Laura; Schaller, Mathias; Tashman, Adam; Toffelmire, Ted; Thijssen, Stephan; Kooman, Jeroen P; van der Sande, Frank M; Levin, Nathan W; Wang, Yuedong; Kotanko, Peter

2013-07-01

332

Subchronic toxicity study in vivo and allergenicity study in vitro for genetically modified rice that expresses pharmaceutical protein (human serum albumin).  

PubMed

Genetically modified (GM) crops that express pharmaceutical proteins have become an important focus of recent genetic engineering research. Food safety assessment is necessary for the commercial development of these crops. Subchronic toxicity study in vivo and allergenicity study in vitro were designed to evaluate the food safety of the rice variety expressing human serum albumin (HSA). Animals were fed rodent diets containing 12.5%, 25.0% and 50.0% GM or non-GM rice for 90 days. The composition analysis of the GM rice demonstrated several significant differences. However, most of the differences remained within the ranges reported in the literature. In the animal study, a range of indexes including clinical observation, feed efficiency, hematology, serum chemistry, organ weights and histopathology were examined. Random changes unrelated to the GM rice exposure, within the range of historical control values and not associated with any signs of illness were observed. The results of heat stability and in vitro digestion of HSA indicated no evidence of potential allergenicity of the protein. Overall, the results of these studies suggest that the GM rice appears to be safe as a dietary ingredient when it is used at up to 50% in the diet on a subchronic basis. PMID:25086369

Sheng, Yao; Qi, Xiaozhe; Liu, Yifei; Guo, Mingzhang; Chen, Siyuan; He, Xiaoyun; Huang, Kunlun; Xu, Wentao

2014-10-01

333

Prolonged Prophylactic Protection from Botulism with a Single Adenovirus Treatment Promoting Serum Expression of a VHH-Based Antitoxin Protein  

PubMed Central

Current therapies for most acute toxin exposures are limited to administration of polyclonal antitoxin serum. We have shown that VHH-based neutralizing agents (VNAs) consisting of two or more linked, toxin-neutralizing heavy-chain-only VH domains (VHHs), each binding distinct epitopes, can potently protect animals from lethality in several intoxication models including Botulinum neurotoxin serotype A1 (BoNT/A1). Appending a 14 amino acid albumin binding peptide (ABP) to an anti-BoNT/A1 heterodimeric VNA (H7/B5) substantially improved serum stability and resulted in an effective VNA serum half-life of 1 to 2 days. A recombinant, replication-incompetent, adenoviral vector (Ad/VNA-BoNTA) was engineered that induces secretion of biologically active VNA, H7/B5/ABP (VNA-BoNTA), from transduced cells. Mice administered a single dose of Ad/VNA-BoNTA, or a different Ad/VNA, via different administration routes led to a wide range of VNA serum levels measured four days later; generally intravenous > intraperitoneal > intramuscular > subcutaneous. Ad/VNA-BoNTA treated mice were 100% protected from 10 LD50 of BoNT/A1 for more than six weeks and protection positively correlated with serum levels of VNA-BoNTA exceeding about 5 ng/ml. Some mice developed antibodies that inhibited VNA binding to target but these mice displayed no evidence of kidney damage due to deposition of immune complexes. Mice were also successfully protected from 10 LD50 BoNT/A1 when Ad/VNA-BoNTA was administered up to 1.5 hours post-intoxication, demonstrating rapid appearance of the protective VNA in serum following treatment. Genetic delivery of VNAs promises to be an effective method of providing prophylactic protection and/or acute treatments for many toxin-mediated diseases. PMID:25170904

Debatis, Michelle; Tremblay, Jacqueline M.; Beamer, Gillian; Kashentseva, Elena A.; Curiel, David T.; Shoemaker, Charles B.

2014-01-01

334

Embryo culture in teratological surveillance and serum proteins in development. Progress report, 1980-1981  

SciTech Connect

The hypothesis that the responses of cultured rat embryos to a medium consisting of serum were indicative of the teratogenic activity of the serum as well as of the teratological risk to the organism donating the serum has been investigated. This hypothesis appeared reasonable because, first, only a whole embryo undergoing rapid growth and development might possess all those events that have the potential of sensitivity to a teratogen. Second, maternal serum has generally been considered a close representation of those substances which might reach the developing embryo. The procedures developed by New (1978) and his associates were used for the isolation of head fold stage (9 1/2 days) rat embryos (with Reichert's membrane removed but yolk sac and ectoplacental cone left intact) and for the preparation of serum as a culture medium (immediate centrifugation and heat inactivation). In addition, we used their recommended 30 rpm culture bottle rotation and their changes of O/sub 2/, CO/sub 2/ and N/sub 2/ levels. All experiments have lasted 48 hours as embryo viability was reduced after this period. The general objective has been to evaluate and to demonstrate possible applications of the cultured rat embryo teratogenic activity test.

Klein, N.W.

1981-09-01

335

The detection of hemorrhagic proteins in snake venoms using monoclonal antibodies against Virginia opossum (Didelphis virginiana) serum.  

PubMed

Most snakes and a few warm-blooded animals have a resistance to snake venoms because of naturally occurring antihemorrhagins found in their sera. The antihemorrhagins in serum of Virginia opossum (Didelphis virginiana) neutralize hemorrhagic activity by binding to hemorrhagins in snake venoms. The binding characteristic of antihemorrhagins in D. virginiana serum was used to develop a five-step western blot. The detection of hemorrhagic proteins were measured indirectly with antihemorrhagins in Virginia opossum serum and with DV-2LD#2, a monoclonal antibody specific for Virginia opossum antihemorrhagins. Snake venoms were separated by native-PAGE, transferred to a Millipore Immobilon-P membrane and then incubated with crude Virginia opossum serum. The hemorrhagins in snake venom bind to antihemorrhagins in Virginia opossum serum which react with DV-2LD#2 a monoclonal antibody that is specific for Virginia opossum antihemorrhagins. DV-2LD#2 monoclonal antibody inhibits antihemorrhagic activity in Virginia opossum serum when mixed in equal amounts. The inhibition of antihemorrhagins by DV-2LD#2 monoclonal antibody suggests specificity. DV-2LD#2 monoclonal antibody does not recognize antihemorrhagins in gray woodrat (Neotoma micropus) serum. The five-step western blot reveals two well-defined bands which represent hemorrhagins found in Western diamondback rattlesnake (Crotalus atrox) venom. Venoms from 15 different snake species were examined to determine the usefulness of the five-step western blot. Other hemorrhagic venoms (Great Basin rattlesnake (C. viridis lutosus), Prairie rattlesnake (C. viridis viridis), Tancitaran dusky rattlesnake (C. pusillus), Northern Mojave rattlesnake (C. scutulatus scutulatus type B) and Northern Pacific rattlesnake (C. v. oreganus)) had one single band in the five-step western blot. DV-2LD#2 did not bind to the non-hemorrhagic venoms and reacted with 50% of the hemorrhagic venoms used in this study. The monoclonal antibody, CAH, reacted with all the hemorrhagic venoms except for the venom of the King cobra (Ophiophagus hannah) and did not react with the non-hemorrhagic venoms. The hemorrhagic binding site of CAH monoclonal antibody and the antihemorrhagin in Virginia opossum are different binding sites. The five-step western blot will be a very useful assay for determining hemorrhagic activity without using live animals. PMID:9723843

Sánchez, E E; García, C; Pérez, J C; De La Zerda, S J

1998-10-01

336

The zinc finger protein A20 protects endothelial cells from burns serum injury.  

PubMed

Burn injuries as well as skin damages are often associated with immune suppression and often cause multiple organ failures. The monolayer endothelium is vulnerable to injuries from circulating factors resulting from remote wounds. Endothelial cell activation and apoptosis can alter microvascular permeability and intensify organ damage. A20, as a physiological cytoprotective gene is essential for preventing spontaneous innate immune cell-mediated inflammation and tissue destruction. It is not known whether A20 has the function to protect endothelial cells from the effect of burns serum challenge on endothelial function in vitro. This study shows that A20 can express in endothelial cells after burns serum stimulation and inhibit endothelial cell activation and apoptosis induced by burns serum. These results suggest that A20 may be beneficial in limiting the response to burn injuries. PMID:15019119

Zhu, Chu-Hong; Ying, Da-Jun; Mi, Jian-Hong; Zhang, Wei; Dong, Shi-Wu; Sun, Jian-Sen; Zhang, Jia-Ping

2004-03-01

337

Gravin, an autoantigen recognized by serum from myasthenia gravis patients, is a kinase scaffold protein  

Microsoft Academic Search

Background: Subcellular targeting of protein kinases and phosphatases provides a mechanism for co-localizing these enzymes with their preferred substrates. A recently identified mammalian scaffold protein, AKAP79, controls the location of two broad-specificity kinases and a phosphatase.Results: We have identified and characterized another mammalian scaffold protein which coordinates the location of protein kinase A and protein kinase C. We isolated a

J. Brian Nauert; Theresa M. Klauck; Lorene K. Langeberg; John D. Scott

1997-01-01

338

The complement control protein homolog of herpesvirus saimiri regulates serum complement by inhibiting C3 convertase activity.  

PubMed Central

The herpesvirus saimiri genome encodes a complement control protein homolog (CCPH). Stable mammalian cell transfectants expressing a recombinant transmembrane form of CCPH (mCCPH) or a 5'FLAG epitope-tagged mCCPH (5'FLAGmCCPH) conferred resistance to complement-mediated cell damage by inhibiting the lytic activity of human serum complement. The function of CCPH was further defined by showing that the mCCPH and the 5'FLAGmCCPH transfectants inhibited C3 convertase activity and effectively reduced cell surface deposition of the activated complement component, C3d. PMID:7745740

Fodor, W L; Rollins, S A; Bianco-Caron, S; Rother, R P; Guilmette, E R; Burton, W V; Albrecht, J C; Fleckenstein, B; Squinto, S P

1995-01-01

339

Electrophoretic Analysis of Blood Serum Proteins in Three Species of Water Snakes (Genus Nerodia)  

Microsoft Academic Search

Serum from three species of water snakes (Nerodia rhombifera, N. erythrogaster and N. fasciata) from one geographic region was analyzed electrophoretically on cellulose acetate, and anodic mobility andrelative concentration of the fractions were determined by a recording densitometer with an automatic integrator. Classification of fractions was based on mobility (R, values), and for identification purposes, bands were labeled in order

PHYLLIS J. GARNETT

340

Concentrations of placental proteins (HPL and SP1) in maternal serum throughout normal pregnancy  

Microsoft Academic Search

Summary Throughout 65 normal singleton pregnancies 332 blood samples were obtained and analyzed for pregnancy-specific ?1 glycoprotein (SP1) and human placental lactogen (HPL). The measurement of SP1 in maternal serum was made using radial immunodiffusion, that of HPL by using radioimmunoassay. There was wide variation in the number and timing of blood samples obtained from patients, and therefore the results

M. P. Baur; O. Bellmann; J. Tebbe; N. Lang

1982-01-01

341

Serum protein layers on parylene-C and silicon oxide: effect on cell adhesion  

E-print Network

and reinforce of the Fn and albumin structural state on these two ls and methods als serum albumin, HPLC grade water (resistiv- cm), phosphate buffered saline (PBS) and sodium lphate (SDS) were purchased from Sigma (Sigma & d used as received. Bovine plasma...

Delivopoulos, Evangelos; Ouberai, Myriam M.; Coffey, Paul D.; Swann, Marcus J.; Shakesheff, Kevin M.; Welland, Mark E.

2014-12-16

342

A Rapid Lateral Flow Immunoassay for the Detection of Tyrosine Phosphatase-Like Protein IA-2 Autoantibodies in Human Serum  

PubMed Central

Type 1 diabetes (T1D) results from the destruction of pancreatic insulin-producing beta cells and is strongly associated with the presence of islet autoantibodies. Autoantibodies to tyrosine phosphatase-like protein IA-2 (IA-2As) are considered to be highly predictive markers of T1D. We developed a novel lateral flow immunoassay (LFIA) based on a bridging format for the rapid detection of IA-2As in human serum samples. In this assay, one site of the IA-2As is bound to HA-tagged-IA-2, which is subsequently captured on the anti-HA-Tag antibody-coated test line on the strip. The other site of the IA-2As is bound to biotinylated IA-2, allowing the complex to be visualized using colloidal gold nanoparticle-conjugated streptavidin. For this study, 35 serum samples from T1D patients and 44 control sera from non-diabetic individuals were analyzed with our novel assay and the results were correlated with two IA-2A ELISAs. Among the 35 serum samples from T1D patients, the IA-2A LFIA, the in-house IA-2A ELISA and the commercial IA-2A ELISA identified as positive 21, 29 and 30 IA-2A-positive sera, respectively. The major advantages of the IA-2A LFIA are its rapidity and simplicity. PMID:25047039

Kikkas, Ingrid; Mallone, Roberto; Larger, Etienne; Volland, Hervé; Morel, Nathalie

2014-01-01

343

A probiotic mixture including galactooligosaccharides decreases fecal ?-glucosidase activity but does not affect serum enterolactone concentration in men during a two-week intervention.  

PubMed

A high serum concentration of enterolactone, an enterolignan produced by colonic microbiota from precursors in cereals, vegetables, and fruits, is associated with reduced risk of acute coronary events. Probiotics and prebiotics modify colonic metabolism and may affect the serum enterolactone concentration. The effects of a probiotic mixture alone and with galactooligosaccharides (GOS) on serum enterolactone concentration and fecal metabolism were investigated in 18 healthy men. Participants received 3 interventions, each for 2 wk: 1) probiotics [Lactobacillus rhamnosus strains GG (LGG) and LC705, Propionibacterium freudenreichii ssp. shermanii JS, and Bifidobacterium breve Bb99, for a total amount of 2 × 10(10) CFU/d]; 2) probiotics and GOS 3.8 g/d; 3) probiotics, GOS, and rye bread (minimum 120 g/d). Serum enterolactone and fecal dry weight, enzyme activities, pH, SCFA, lactic acid bacteria, bifidobacteria, propionibacteria, and the strains LGG and LC705 were determined. The serum enterolactone concentration (nmol/L) tended to be decreased from baseline [mean (95% CI) 18.6 (10.8-26.4)] by probiotics alone [15.2 (7.8-22.7); P = 0.095], was not significantly affected by probiotics with GOS [21.5 (13.2-29.8)], and was increased by probiotics with GOS and rye bread [24.6 (15.4-33.7); P < 0.05]. Probiotics alone did not affect fecal ?-glucosidase activity and bifidobacteria, but probiotics with GOS decreased ?-glucosidase activity and increased bifidobacteria compared with baseline (P < 0.05) and with probiotics alone (P < 0.01). In conclusion, this probiotic mixture with or without GOS does not significantly affect serum enterolactone concentration. Because probiotics with GOS decreased fecal ?-glucosidase activity but not serum enterolactone, the reduced fecal ?-glucosidase, within the range of activities measured, does not seem to limit the formation of enterolactone. PMID:21411613

Kekkonen, Riina A; Holma, Reetta; Hatakka, Katja; Suomalainen, Tarja; Poussa, Tuija; Adlercreutz, Herman; Korpela, Riitta

2011-05-01

344

Osmotically unresponsive water fraction on proteins: Non-ideal osmotic pressure of bovine serum albumin as a function of pH and salt concentration  

Microsoft Academic Search

How much does protein-associated water differ in colligative properties (freezing point, boiling point, vapor pressure and osmotic behavior) from pure bulk water? This question was approached by studying the globular protein bovine serum albumin (BSA), using changes in pH and salt concentration to alter its native structural conformation and state of aggregation. BSA osmotic pressure was investigated experimentally and analyzed

Gary D. Fullerton; Kalpana M. Kanal; Ivan L. Cameron

2006-01-01

345

Placental Lactogen Administration Reverses the Effect of Low-Protein Diet on Maternal and Fetal Serum Somatomedin Levels in the Pregnant Rat  

Microsoft Academic Search

Female rats were studied on day 20 of pregnancy after being fed either a 5% lactalbumin (low protein) diet or a 20% lactalbumin (adequate) diet for the last 2 weeks of pregnancy. Rats on the lower intake of protein showed decreased serum levels of rat placental lactogen and reduced numbers of lactogenic receptors in the maternal liver. These changes were

S. J. Pilistine; A. C. Moses; H. N. Munro

1984-01-01

346

The diagnostic role of human epididymis protein 4 and serum amyloid-A in early-stage endometrial cancer patients.  

PubMed

The aim of this study was to evaluate the prognostic and predictive efficacy of the human epididymis secretory protein 4 (HE4) and serum amyloid-A (S-AA) together with the other tumor markers (CA 125, CA 15-3, CEA, and CA 19-9) in endometrial cancer patients. The study group consisted of 64 patients with defined stage and grade of endometrial cancer and 60 women with benign uterine diseases. Thirty-four healthy women were defined as the control group. Fasting blood samples were collected prior to surgery and tumor marker levels were determined in blood samples by E170 autoanalyzer. S-AA concentrations were measured by particle-enhanced immunonephelometry. Preoperative serum HE4 and S-AA levels were significantly higher in endometrial cancer patients than in controls, whereas the other measured parameters were not significantly different. Serum levels of HE4 were related to both the stage and grade of tumor. The best cutoff point for HE4 was determined to be 59.7 pmol/L; with 75 % sensitivity and 65.5 % specificity. For S-AA, the cutoff point was 8.8 U/mL, with 68.7 % sensitivity and 58.6 % specificity. The combination of HE4, CA 125, CEA, and S-AA raised the sensitivity to 84 %. Preoperative measurement of serum HE4 and S-AA may be of help in early detection of endometrial cancer. Preoperative screening with these markers may provide important information about the patient's outcome and prognosis. PMID:23640061

Omer, Beyhan; Genc, Sema; Takmaz, Ozguc; Dirican, Ahmet; Kusku-Kiraz, Zeynep; Berkman, Sinan; Gurdol, Figen

2013-10-01

347

Selective binding of naphthoquinone derivatives to serum albumin proteins and their effects on cytotoxicity.  

PubMed

Naphthoquinone derivatives such as lapachol, plumbagin, dichloroallyl lawsone show anticancer activity and generally cytotoxicity measurements are carried out in presence of bovine serum albumin; so understanding on the ability of serum albumin binding with such derivatives are essential. We have investigated cytotoxicity and serum albumin binding of a series of structurally related naphthoquinone derivatives. Substrate dependency and high selectivity in binding of naphthoquinone tethered carboxylic acids or pyridines with bovine serum albumin (BSA) and human serum albumin (HSA) are observed. For example, the binding constant of BSA with 3-(1,4-dihydro-2-methyl-1,4-dioxonaphthalen-3yl-thio)propanoic acid is ?594 times higher than 3-(1,4-dioxo-1,4-dihydronaphthalen-2-yl-amino)benzoic acid; whereas 4-(1,4-dioxo-1,4-dihydronaphthalen-2-yl-amino)benzoic acid shows ?367 times higher binding constant than the latter compound. The BSA weakly bind to pyridine tethered naphthoquinones, whereas HSA does not binds with them. The binding constant of HSA with 2-(1,4-dihydro-2-methyl-1,4-dioxonaphthalene-3-ylthio)benzoic acid is 134 times higher than the HSA binding constant with 2,2'-(1,4-dihydro-1,4-dioxo-naphthalen-2,3-diylthio)dipropanoic acid. Among the naphthoquinone carboxylic acids, the 3-(1,4-dioxo-1,4-dihydronaphthalen-2-yl-amino)benzoic acid binds selectively to BSA, but it does not bind to HSA. The 2-hydroxybenzoic acid or 4-mercaptobenzoic acid strongly binds to BSA. The binding of BSA with 4-hydroxybenzoic acid or 2-mercaptobenzoic acid are insignificant. We have not observed clear relationships of structure of naphthoquinone derivatives versus serum albumin binding, but could identify the compound having the best IC50 values of cytotoxicity among the twelve naphthoquinone compounds. The compound 3-(1,2-dihydro-1,2-dioxonaphthalen-4-yl-thio)propanoic acid in four cancer cell lines has IC50 values in the range 2.7-7.6?M. This compound also has optimum binding constant with BSA (35.042×10(3)Lmol(-1)) or HSA (21.427×10(3)Lmol(-1)). The cytotoxicity values of the compounds were influenced by concentration of BSA. PMID:24560625

Jali, Bigyan R; Kuang, Yuting; Neamati, Nouri; Baruah, Jubaraj B

2014-05-01

348

Exploring the interaction of a micelle entrapped biologically important proton transfer probe with the model transport protein bovine serum albumin.  

PubMed

This article describes the interaction of a micelle entrapped pharmaceutically important isoindole fused imidazole derivative, namely, 1-(2-hydroxy-5-methyl-phenyl)-3,5-dioxo-1H-imidazo-[3,4-b] isoindole (ADII), with the model transport protein bovine serum albumin (BSA). Different spectroscopic techniques such as steady state absorption, emission, circular dichroism, dynamic light scattering, etc., have been employed to explore preferential interaction of this drug template with micelles and protein BSA. Binding of ADII with BSA is found to be enormously modified when it is released from the micellar environment. The binding constant of the ADII-BSA complex is reduced when the probe is released from anionic SDS micelle, whereas the binding is observed to be strengthened in cationic CTAB micellar medium due to the formation of a 1:2 complex (ADII-BSA). Time-resolved studies also support our steady state findings that the released drug from the micellar environment is found to be strongly bound with the protein BSA. Circular dichroism (CD) and dynamic light scattering (DLS) study reveals that the secondary structure of BSA gets some stabilization in SDS medium after binding of drug template to protein. The probable binding location of the probe within the protein cavity (hydrophilic subdomain IA) has been explored from an AutoDock-based blind docking simulation study. PMID:25068392

Ray, Debarati; Kundu, Ashis; Pramanik, Animesh; Guchhait, Nikhil

2015-02-12

349

A novel protein from the serum of Python sebae, structurally homologous with type-? phospholipase A(2) inhibitor, displays antitumour activity.  

PubMed

Cytotoxic and antitumour factors have been documented in the venom of snakes, although little information is available on the identification of cytotoxic products in snake serum. In the present study, we purified and characterized a new cytotoxic factor from serum of the non-venomous African rock python (Python sebae), endowed with antitumour activity. PSS (P. sebae serum) exerted a cytotoxic activity and reduced dose-dependently the viability of several different tumour cell lines. In a model of human squamous cell carcinoma xenograft (A431), subcutaneous injection of PSS in proximity of the tumour mass reduced the tumour volume by 20%. Fractionation of PSS by ion-exchange chromatography yielded an active protein fraction, F5, which significantly reduced tumour cell viability in vitro and, strikingly, tumour growth in vivo. F5 is composed of P1 (peak 1) and P2 subunits interacting in a 1:1 stoichiometric ratio to form a heterotetramer in equilibrium with a hexameric form, which retained biological activity only when assembled. The two peptides share sequence similarity with PIP {PLI-? [type-? PLA(2) (phospholipase A(2)) inhibitor] from Python reticulatus}, existing as a homohexamer. More importantly, although PIP inhibits the hydrolytic activity of PLA(2), the anti-PLA(2) function of F5 is negligible. Using high-resolution MS, we covered 87 and 97% of the sequences of P1 and P2 respectively. In conclusion, in the present study we have identified and thoroughly characterized a novel protein displaying high sequence similarity to PLI-? and possessing remarkable cytotoxic and antitumour effects that can be exploited for potential pharmacological applications. PMID:21834793

Donnini, Sandra; Finetti, Federica; Francese, Simona; Boscaro, Francesca; Dani, Francesca R; Maset, Fabio; Frasson, Roberta; Palmieri, Michele; Pazzagli, Mario; De Filippis, Vincenzo; Garaci, Enrico; Ziche, Marina

2011-12-01

350

Contractile protein gene expression in serum-free cultured adult rat cardiac myocytes.  

PubMed

The effects of two adhesion substrates (serum and laminin) and time in culture on the expression of genes encoding myosin heavy chain (MHC) isoforms and alpha-skeletal actin were analysed in myocytes isolated from adult rat heart and maintained in serum-free culture. Relative messenger ribonucleic acid (mRNA) abundances were quantitated by dot-blot analysis. Gene expression was not influenced by the substrate used. Time in culture induced a decrease in total mRNA abundance and an up-regulation of beta-MHC and alpha-skeletal actin genes. It is proposed that atrophy of adult myocytes is associated with a pattern of gene expression similar to the fetal program. PMID:8351198

Dubus, I; Rappaport, L; Barrieux, A; Lompré, A M; Schwartz, K; Samuel, J L

1993-06-01

351

Population genetic studies of the Philippine Negritos. I. A pilot survey of red cell enzyme and serum protein groups.  

PubMed Central

Electrophoretic surveys of red cell enzyme and serum protein systems representing 21 genetic loci were carried out on 129 blood samples of the Negritos of Pampanga, Central Luzon, the Philippines. Nine (out of 16) red cell enzyme loci and four (out of five) serum protein loci showed polymorphic variation. Low frequencies of ACP 1A, GPTs1, ESD2, and Hp1, and a markedly high frequency of PGM12 were contrasted to those in non-Negrito Filipinos. Variant ESD phenotypes with a slowly migrating isozyme occurred in high frequency. The new allele designated as ESD3Negrito (ESD3N) had a frequency of .10 +/- .019. In AK, a variant phenotype indistinguishable from AK 2-1 was observed in 14% of the sample. In the Gc system, a fast migrating variant was discovered in high frequency which was distinct from Gc Ab and Gc J. The variant allele, denoted GcNegrito (GcN), had a frequency of .21 +/- .025. A relatively high degree of allelic diversity in the Negrito sample was also suggested by the average heterozygosity for 21 loci screened (.165), which is compared to that of the Japanese population (.140). Images Fig. 2 Fig. 3 PMID:655166

Omoto, K; Misawa, S; Harada, S; Sumpaico, J S; Medado, P M; Ogonuki, H

1978-01-01

352

Myeloperoxidase and eosinophil cationic protein in serum and sputum during antibiotic treatment in cystic fibrosis patients with Pseudomonas aeruginosa infection  

PubMed Central

I order to study the time-course of myeloperoxidase (MPO) and eosinophil cationic protein (ECP) as parameters for monitoring inflammation in cystic fibrosis (CF), we investigated ten patients during both a 14-day intravenous antibiotic treatment and a corresponding self control. Modified Shwachman-Kulczycki score improved significantly (p < 0.008), C-reactive protein (CRP) levels decreased significantly (p < 0.05) during antibiotic treatment, while in the control phase there were no significant differences. Lung function parameters did not change significantly during antibiotic treatment or control phase. Serum MPO concentration (p < 0.006) and peripheral blood neutrophil granulocyte counts (p < 0.04) decreased significantly during antibiotic treatment, but not during the control phase. Sentm ECP concentration showed a tendency to decrease during antibiotic treatment, but this failed to reach significance. In general, sputum concentrations of MPO and ECP Were 500- to 1000-fold higher than in serum. However, neither MPO nor ECP in sputum showed a significan variation over time during antibiotic treatment or control phase. From our data we conclude that: (1) measurements of MPO, neutrophils and CRP in peripheral blood do correlate with clinical parameters such as the modified Shwachman-Kulczycki score; (2) neutrophils and MPO seem to reflect inflammatory changes induced by antibiotic treatment; (3) eosinophils may play a role in CF by an enhanced ‘releasability’ and (4) Sputum measurements of mediators of inflammation cannot be recommended. PMID:18475652

Stiller, T.; Magdorf, K.; Wahn, U.

1995-01-01

353

Acute-phase protein serum amyloid A3 is a novel paracrine coupling factor that controls bone homeostasis.  

PubMed

Serum amyloid A (A-SAA/Saa3) was shown before to affect osteoblastic metabolism. Here, using RT-quantitative PCR and/or immunoblotting, we show that expression of mouse Saa3 and human SAA1 and SAA2 positively correlates with increased cellular maturation toward the osteocyte phenotype. Expression is not detected in C3H10T1/2 embryonic fibroblasts but is successively higher in preosteoblastic MC3T3-E1 cells, late osteoblastic MLO-A5 cells, and MLO-Y4 osteocytes, consistent with findings using primary bone cells from newborn mouse calvaria. Recombinant Saa3 protein functionally inhibits osteoblast differentiation as reflected by reductions in the expression of osteoblast markers and decreased mineralization in newborn mouse calvaria. Yet, Saa3 protein enhances osteoclastogenesis in mouse macrophages/monocytes based on the number of multinucleated and tartrate-resistant alkaline phosphatase-positive cells and Calcr mRNA expression. Depletion of Saa3 in MLO osteocytes results in the loss of the mature osteocyte phenotype. Recombinant osteocalcin, which is reciprocally regulated with Saa3 at the osteoblast/osteocyte transition, attenuates Saa3 expression in MLO-Y4 osteocytes. Mechanistically, Saa3 produced by MLO-Y4 osteocytes is integrated into the extracellular matrix of MC3T3-E1 osteoblasts, where it associates with the P2 purinergic receptor P2rx7 to stimulate Mmp13 expression via the P2rx7/MAPK/ERK/activator protein 1 axis. Our data suggest that Saa3 may function as an important coupling factor in bone development and homeostasis.-Thaler, R., Sturmlechner, I., Spitzer, S., Riester, S. M., Rumpler, M., Zwerina, J., Klaushofer, K., van Wijnen, A. J., and Varga, F. Acute-phase protein serum amyloid A3 is a novel paracrine coupling factor that controls bone homeostasis. PMID:25491310

Thaler, Roman; Sturmlechner, Ines; Spitzer, Silvia; Riester, Scott M; Rumpler, Monika; Zwerina, Jochen; Klaushofer, Klaus; van Wijnen, Andre J; Varga, Franz

2014-12-01

354

Blood–brain barrier damage and brain penetration of antiepileptic drugs: Role of serum proteins and brain edema  

PubMed Central

SUMMARY Purpose: Increased blood–brain barrier (BBB) permeability is radiologically detectable in regions affected by drug-resistant epileptogenic lesions. Brain penetration of antiepileptic drugs (AEDs) may be affected by BBB damage. We studied the effects of BBB damage on brain distribution of hydrophilic [deoxy-glucose (DOG) and sucrose] and lipophilic (phenytoin and diazepam) molecules. We tested the hypothesis that lipophilic and hydrophilic drug distribution is differentially affected by BBB damage. Methods: In vivo BBB disruption (BBBD) was performed in rats by intracarotid injection of hyperosmotic mannitol. Drugs (H3-sucrose, 3H-deoxy-glucose, 14C-phenytoin, and C14-diazepam) or unlabeled phenytoin was measured and correlated to brain water content and protein extravasation. In vitro hippocampal slices were exposed to different osmolarities; drug penetration and water content were assessed by analytic and densitometric methods, respectively. Results: BBBD resulted in extravasation of serum protein and radiolabeled drugs, but was associated with no significant change in brain water. Large shifts in water content in brain slices in vitro caused a small effect on drug penetration. In both cases, total drug permeability increase was greater for lipophilic than hydrophilic compounds. BBBD reduced the amount of free phenytoin in the brain. Discussion: After BBBD, drug binding to protein is the main controller of total brain drug accumulation. Osmotic BBBD increased serum protein extravasation and reduced free phenytoin brain levels. These results underlie the importance of brain environment and BBB integrity in determining drug distribution to the brain. If confirmed in drug-resistant models, these mechanisms could contribute to drug brain distribution in refractory epilepsies. PMID:19175391

Marchi, Nicola; Betto, Giulia; Fazio, Vincent; Fan, Quinyuan; Ghosh, Chaitali; Machado, Andre; Janigro, Damir

2009-01-01

355

Embryo culture in teratological surveillance and serum proteins in development. Progress report, 1978-1979  

Microsoft Academic Search

Two initial studies on the use of cultured rat embryos for teratological surveillance have been completed using cadmium and cyclophosphamide teratogens. It was found that rat embryos could be cultured successfully on human serum supplemented with glucose, allowing testing to extend to human subjects. Tests using energy-related fuel substances (petroleum crude; shale oil; two coal-derived oils) implied possible long-term growth-inhibition

1979-01-01

356

Tubule urate and PAH transport: sensitivity and specificity of serum protein inhibition  

Microsoft Academic Search

Macromolecules in rabbit serum inhibit the cellular uptake and transepithelial secretion of (¹⁴C)urate and p-(³H)aminohippurate ((³H)PAH) in rabbit Sâ proximal tubule segments. To understand better the potential role these inhibitors may have in the regulation of renal organic anion excretion, the authors examined the specificity and relative inhibitory effects on tubule urate and PAH transport of albumin and ..gamma..-globulin, the

J. J. Grantham; J. Kennedy; B. Cowley

1987-01-01

357

Determination of C?reactive protein in serum and plasma from healthy dogs and dogs with pneumonia by ELISA and slide reversed passive latex agglutination test  

Microsoft Academic Search

The concentrations of C?reactive protein (CRP) in serum from dogs diagnosed as normal by clinical, haematological, and biochemical examination were determined by enzyme?linked immunosorbent assay (ELISA) and slide reversed passive latex agglutination (RPLA), using IgG antibody isolated from rabbit anti?canine CRP serum. The mean value of CRP in 66 normal dogs kept in private households was 8.4± 4.9 ?g\\/ml by

S. Yamamoto; T. Shida; T. Okimura; K. Otabe; M. Honda; Y. Ashida; E. Furukawa; M. Sarikaputi; M. Naiki

1994-01-01

358

Total internal reflected resonance light scattering determination of protein in human blood serum at water\\/tetrachloromethane interface with Arsenazo-TB and Cetyltrimethylammonium bromide  

Microsoft Academic Search

A direct quantification of protein in human blood serum samples is proposed based on the measurement of total internal reflected resonance light scattering (TIR-RLS) at water\\/tetrachloromethane (H2O\\/CCl4) interfaces. At pH 4.10 and in the presence of Cetyltrimethylammonium bromide (CTAB), the interaction of Arsenazo-TB (ATB) with human serum albumin (HSA) results in strong enhanced TIR-RLS signals with the maximum spectra peak

Lijun Dong; Xingguo Chen; Zhide Hu

2007-01-01

359

Alternative pathway fof bovine complement Immunochemical studies on factor B-like serum protein and its conversion product B gamma 2.  

PubMed Central

Rabbits produced antibodies to a factor B-like serum protein (factor Bbov), its conversion product B gamma 2 and some other bovine serum proteins after repeated immunization with zymosan which previously had been incubated with fresh bovine serum. Such antisera were used to monitor purification of B gamma 2 from fresh bovine sera incubated with zxymosan. Subsequently, antisera specific for factor Bbov and B gamma 2 were produced. Antiserum produced against B gamma 2 cross-reacted with factor Bbov. Functional assays for factor Bbov were carried out in a hemolytic system with guinea pig erythrocytes in EGTA buffer. Heat inactivation (56 degrees C/5 min) of bovine serum destroyed the antigenicity of factor Bbov but not that of B gamma 2. Factor Bbov had an apparent molecular weight of 95,000 and B gamma 2 a molecular weight of 40,000 daltons. Conversion of factor Bbov to B gamma 2 was determined qualitatively by immunoelectrophoresis and quantitatively by radial immunodiffusion. Conversion of factor Bbov to B gamma 2 in bovine serum, in the presence of zymosan or cobra venom factor (CoVF) required Mg++ but not Ca++, did not occur in heat inactivated (56 degrees C/5 min) serum and was maximal, but not complete, when fresh bovine serum was incubated with zymosan (20 mg/mL) at 37 for two hours. Images Fig. 1. Fig. 2 Fig. 3. Fig. 4. Fig. 6. Fig. 7. PMID:6176300

Tabel, H

1981-01-01

360

Investigation of the cumulative tissue doses of naphthoquinones in human serum using protein adducts as biomarker of exposure.  

PubMed

Both 1,2-naphthoquinone (1,2-NPQ) and 1,4-naphthoquinone (1,4-NPQ) are reactive metabolites of naphthalene that are thought to be responsible for the naphthalene-induced cytotoxicity and genotoxicity. The aim of this study was to investigate the cumulative tissue dose of 1,2-NPQ and 1,4-NPQ in human serum derived from blood donors in Taiwan via measurements of albumin adducts by a methodology, which employs trifluoroacetic acid anhydride and methanesulfonic acid to selectively cleave cysteinyl adducts on proteins. Both 1,2-NPQ and 1,4-NPQ adducts were detected in all male and female subjects (n=22). The median levels of 1,2-NPQ adduct in human subjects were estimated to be 268 (range 139-857) and 203 (range 128-1352) (pmol/g) in male (n=11) and female (n=11) subjects, respectively. In contrast, the median levels of 1,4-NPQ adduct were estimated to be 45.0 (range 22.0-117) and 38.9 (range 21.5-172) (pmol/g) in male and female subjects, respectively. We noticed that levels of 1,2-NPQ adduct were significantly correlated with those of 1,4-NPQ adduct (correlation coefficient r=0.643, p<0.01). Results from in vitro experiments confirmed that the production of naphthoquinones-derived adducts on serum albumin increased with increased concentration of naphthoquinones (0-100 microM). Linear relationships were observed over the range of concentration. Time-course experiments suggested that both 1,2-NPQ and 1,4-NPQ-derived adducts rapidly reached maximum values at 10 min mark and remained constant thereafter. The reaction rate constant analyses indicated that the second-order rate constants, representing in vitro reactions between naphthoquinones and cysteine residues of serum albumin, were estimated to be 0.0044/0.0002L(gprotein)(-1)h(-1), respectively. Overall, the cumulative tissue doses of 1,4-NPQ (217-316 nM h) in male and female subjects were approximately 3-fold greater than those of 1,2-NPQ (76-98 nM h) in the study population. The initial concentrations of serum 1,2-NPQ and 1,4-NPQ in the study population were estimated to be between 145-188 and 807-1175 nM, respectively. We conclude that the relatively large amounts of naphthoquinones present in human serum may point to toxicological consequences. PMID:19505452

Lin, Po-Hsiung; Chen, Dar-Ren; Wang, Tzu-Wen; Lin, Chia-Hua; Chuang, Ming-Chieh

2009-09-14

361

Maternal rat serum concentrations of dimethadione do not explain intra-litter differences in the incidence of dimethadione-induced birth defects, including novel findings in foetal lung.  

PubMed

To investigate mechanisms of chemical-induced congenital heart defects (CHD) we have developed a rat model using dimethadione (DMO), the N-demethylated metabolite of the anticonvulsant, trimethadione (TMD). Dosing pregnant rats with 300mg/kg DMO every 12h from the evening of gestational day (GD) 8 until the morning of GD 11 (six total doses) produces a mean 74% incidence of CHD with inter litter variability ranging from 40 to 100%. The goal of this study was to determine if the variability in maternal serum concentrations of DMO on GD 14, a surrogate marker for total exposure, was related to the inter-litter differences in teratogenic outcomes. To test this hypothesis, pregnant rats were dosed as described above and serum levels of DMO assessed on GD 14. On GD 21, foetuses were collected by caesarean section, assessed for a number endpoints and the outcomes were correlated with the GD 14 serum concentrations of DMO. DMO exposure was associated with decreased foetal body weight, increased incidence of sternal defects and CHD, but these endpoints were not meaningfully correlated with maternal concentrations of DMO. Novel findings were decreased viability as measured one-hour following caesarean section, and delayed alveolar maturation. The major conclusions from these studies were first, that serum DMO concentrations on GD 14 did not predict teratogenicity, and second, delayed lung development may contribute to the decreased survival of foetuses at the time of caesarean section. PMID:25446330

Rodger, Ian; Lam, Isabel; Purssell, Elizabeth; Thompson, Mesha; Rutter, Allison; Ozolinš, Terence Rs

2014-12-01

362

Structural analysis of sulphated glycoprotein 2 from amino acid sequence. Relationship to clusterin and serum protein 40,40.  

PubMed Central

Sulphated glycoprotein 2 (SGP-2) is the major secreted protein product of rat Sertoli cells; likewise, clusterin is a major constituent of ram rete testis fluid. Isolation and sequencing of the intact subunits and peptides derived from clusterin show that it is the ram homologue of rat SGP-2. Human serum protein 40,40 (SP-40,40), a component of the SC5b-9 complex of complement, has recently been reported to be the human homologue of rat SGP-2. Analysis of the amino acid sequences of rat SGP-2 and human SP-40,40 show that both of these proteins have a significant relationship to the heavy chain of myosin. The regions of highest sequence similarity correspond to the major amphipathic domains in SGP-2/SP-40,40 and the long alpha-helical-tail domain of myosin, which forms a rod-like structure. SGP-2 has anomalous sedimentation behaviour which indicates that it probably exists in an extended conformation. A putative dinucleotide-binding structure has been identified in the longest stretch of identity between SGP-2 and SP-40,40. Elucidation of these features of SGP-2 and SP-40,40 may help to direct future studies into the role of these proteins in the reproductive and complement systems. Images Fig. 4. PMID:2363694

Tsuruta, J K; Wong, K; Fritz, I B; Griswold, M D

1990-01-01

363

In-vitro Galactation of Human Serum Albumin: Analysis of the Protein’s Galactation Sites by Mass Spectrometry  

PubMed Central

The post-translational modification of proteins by sugars has been demonstrated in diabetes and classical galactosemia. In diabetes, the glycation process occurs as a result of D-glucose nonenzymatically reacting with proteins such as albumin and hemoglobin, used today as important tools to monitor the efficiency of dietary control and therapy during treatment of diabetes. In classical galactosemia, D-galactose contributes to the formation of glycated proteins also, suggesting that akin to diabetes with glucated proteins, the monitoring of galactated proteins may facilitate management of patients with galactosemia. The objectives of this project were: 1) galactate HSA, in vitro; 2) determine, by a sodium borohydride dependent mass peptide mapping method, the galactation sites in HSA; and 3) compare HSA’s galactation sites with the protein’s reported glucation sites. Treatment of galactated HSA with sodium borohydride stabilized the condensed sugars on the protein and yielded discrete fragmentation patterns by tandem MS, allowing reliable identification of HSA’s galactation sites. LC/ESI/MS, in combination with tandem MS, revealed the principal sites of galactation in HSA were the amino groups of lysine 12, 233, 281/276, 414 and 525. Lysyl residues 12, 233, 276, and 525 were previously reported as privileged sites for the nonenzymatic binding of D-glucose with HSA. PMID:21112314

Leslie, Frost; Muhammad, Chaudhry; Tiffany, Bell; Menashi, Cohenford

2010-01-01

364

Constraint-based Local Move Definitions for Lattice Protein Models Including Side Chains  

Microsoft Academic Search

The simulation of a protein's folding process is often done via stochastic\\u000alocal search, which requires a procedure to apply structural changes onto a\\u000agiven conformation. Here, we introduce a constraint-based approach to enumerate\\u000alattice protein structures according to k-local moves in arbitrary lattices.\\u000aOur declarative description is much more flexible for extensions than standard\\u000aoperational formulations. It enables a

Martin Mann; Mohamed Abou Hamra; Kathleen Steinhöfel; Rolf Backofen

2009-01-01

365

The Importance of Protein-Protein Interactions on the pH-Induced Conformational Changes of Bovine Serum Albumin: A Small-Angle X-Ray Scattering Study  

PubMed Central

Abstract The combined effects of concentration and pH on the conformational states of bovine serum albumin (BSA) are investigated by small-angle x-ray scattering. Serum albumins, at physiological conditions, are found at concentrations of ?35–45 mg/mL (42 mg/mL in the case of humans). In this work, BSA at three different concentrations (10, 25, and 50 mg/mL) and pH values (2.0–9.0) have been studied. Data were analyzed by means of the Global Fitting procedure, with the protein form factor calculated from human serum albumin (HSA) crystallographic structure and the interference function described, considering repulsive and attractive interaction potentials within a random phase approximation. Small-angle x-ray scattering data show that BSA maintains its native state from pH 4.0 up to 9.0 at all investigated concentrations. A pH-dependence of the absolute net protein charge is shown and the charge number per BSA is quantified to 10(2), 8(1), 13(2), 20(2), and 26(2) for pH values 4.0, 5.4, 7.0, 8.0, and 9.0, respectively. The attractive potential diminishes as BSA concentration increases. The coexistence of monomers and dimers is observed at 50 mg/mL and pH 5.4, near the BSA isoelectric point. Samples at pH 2.0 show a different behavior, because BSA overall shape changes as a function of concentration. At 10 mg/mL, BSA is partially unfolded and a strong repulsive protein-protein interaction occurs due to the high amount of exposed charge. At 25 and 50 mg/mL, BSA undergoes some re-folding, which likely results in a molten-globule state. This work concludes by confirming that the protein concentration plays an important role on the pH-unfolded BSA state, due to a delicate compromise between interaction forces and crowding effects. PMID:20085727

Barbosa, Leandro R.S.; Ortore, Maria Grazia; Spinozzi, Francesco; Mariani, Paolo; Bernstorff, Sigrid; Itri, Rosangela

2010-01-01

366

A highly sensitive "turn-on" fluorescent sensor for the detection of human serum proteins based on the size exclusion of the polyacrylamide gel.  

PubMed

A highly sensitive "turn-on" fluorescent sensor based on the size exclusion of the polyacrylamide gel was developed for the on-gels detection of human serum proteins after PAGE. The possible mechanism of this fluorescence sensor was illustrated and validated by utilizing five kinds of colloidal silver nanoparticles with different particle size distribution and six kinds of polyacrylamide gels with different pore size. It was attributed to that silver nanoparticles (<5 nm in diameter) had been selectively absorbed into the gel and formed the small silver nanoclusters, resulting in the red fluorescence. Using this new technique for the detection of human serum proteins after PAGE, a satisfactory sensitivity was achieved and some relatively low-abundance proteins (e.g. zinc-alpha-2-glycoprotein), which are the significant proteinic markers of certain diseases can be easily detected, but not with traditional methods. Furthermore, it was also successfully applied to distinguish between serums from hepatoma patient and healthy people. As a new protein detection technique, the colloidal silver nanoparticles based "turn-on" fluorescent sensor offers a rapid, economic, low background, and sensitive way for direct detection of human serum proteins, showing available potential and significance in the development of nanobiotechnology and proteome research. PMID:24150987

Xu, Shenghao; Liu, Pingping; Lu, Xin; Zhang, Jing; Huang, Lingyun; Hua, Wenhao; He, Dacheng; Ouyang, Jin

2014-02-01

367

Small serum protein-1 changes the susceptibility of an apoptosis-inducing metalloproteinase HV1 to a metalloproteinase inhibitor in habu snake (Trimeresurus flavoviridis)  

PubMed Central

Viperidae snakes containing various venomous proteins also have several anti-toxic proteins in their sera. However, the physiological function of serum protein has been elucidated incompletely. Small serum protein (SSP)-1 is a major component of the SSPs isolated from the serum of a Japanese viper, the habu snake (Trimeresurus flavoviridis). It exists in the blood as a binary complex with habu serum factor (HSF), a snake venom metalloproteinase inhibitor. Affinity chromatography of the venom on an SSP-1-immobilized column identified HV1, an apoptosis-inducing metalloproteinase, as the target protein of SSP-1. Biacore measurements revealed that SSP-1 was bound to HV1 with a dissociation constant of 8.2 × 10?8 M. However, SSP-1 did not inhibit the peptidase activity of HV1. Although HSF alone showed no inhibitory activity or binding affinity to HV1, the SSP-1–HSF binary complex bound to HV1 formed a ternary complex that non-competitively inhibited the peptidase activity of HV1 with a inhibition constant of 5.1 ± 1.3 × 10?9 M. The SSP-1–HSF complex also effectively suppressed the apoptosis of vascular endothelial cells and caspase 3 activation induced by HV1. Thus, SSP-1 is a unique protein that non-covalently attaches to HV1 and changes its susceptibility to HSF. PMID:23100271

Shioi, Narumi; Ogawa, Eiki; Mizukami, Yuki; Abe, Shuhei; Hayashi, Rieko; Terada, Shigeyuki

2013-01-01

368

Increased Serum and Musculotendinous Fibrogenic Proteins following Persistent Low-Grade Inflammation in a Rat Model of Long-Term Upper Extremity Overuse  

PubMed Central

We examined the relationship between grip strength declines and muscle-tendon responses induced by long-term performance of a high-repetition, low-force (HRLF) reaching task in rats. We hypothesized that grip strength declines would correlate with inflammation, fibrosis and degradation in flexor digitorum muscles and tendons. Grip strength declined after training, and further in weeks 18 and 24, in reach limbs of HRLF rats. Flexor digitorum tissues of reach limbs showed low-grade increases in inflammatory cytokines: IL-1? after training and in week 18, IL-1? in week 18, TNF-? and IL-6 after training and in week 24, and IL-10 in week 24, with greater increases in tendons than muscles. Similar cytokine increases were detected in serum with HRLF: IL-1? and IL-10 in week 18, and TNF-? and IL-6 in week 24. Grip strength correlated inversely with IL-6 in muscles, tendons and serum, and TNF-? in muscles and serum. Four fibrogenic proteins, TGFB1, CTGF, PDGFab and PDGFbb, and hydroxyproline, a marker of collagen synthesis, increased in serum in HRLF weeks 18 or 24, concomitant with epitendon thickening, increased muscle and tendon TGFB1 and CTGF. A collagenolytic gelatinase, MMP2, increased by week 18 in serum, tendons and muscles of HRLF rats. Grip strength correlated inversely with TGFB1 in muscles, tendons and serum; with CTGF-immunoreactive fibroblasts in tendons; and with MMP2 in tendons and serum. Thus, motor declines correlated with low-grade systemic and musculotendinous inflammation throughout task performance, and increased fibrogenic and degradative proteins with prolonged task performance. Serum TNF-?, IL-6, TGFB1, CTGF and MMP2 may serve as serum biomarkers of work-related musculoskeletal disorders, although further studies in humans are needed. PMID:24015193

Gao, Helen G. L.; Fisher, Paul W.; Lambi, Alex G.; Wade, Christine K.; Barr-Gillespie, Ann E.; Popoff, Steven N.; Barbe, Mary F.

2013-01-01

369

C-reactive protein, haptoglobin, serum amyloid A and pig major acute phase protein response in pigs simultaneously infected with H1N1 swine influenza virus and Pasteurella multocida  

PubMed Central

Background Swine influenza (SI) is an acute respiratory disease caused by swine influenza virus (SIV). Swine influenza is generally characterized by acute onset of fever and respiratory symptoms. The most frequent complications of influenza are secondary bacterial pneumonia. The objective of this work was to study the acute phase proteins (APP) responses after coinfection of piglets with H1N1 swine influenza virus (SwH1N1) and Pasteurella multocida (Pm) in order to identify whether the individual APP response correlate with disease severity and whether APP could be used as markers of the health status of coinfected pigs. Results In all coinfected pigs clinical sings, including fever, coughing and dyspnea, were seen. Viral shedding was observed from 2 to 7 dpi. The mean level of antibodies against Pm dermonecrotoxin in infected piglets increase significantly from 7 dpi. Anti-SwH1N1 antibodies in the serum were detected from 7 dpi. The concentration of C-reactive protein (CRP) increased significantly at 1 dpi as compared to control pigs, and remained significantly higher to 3 dpi. Level of serum amyloid A (SAA) was significantly higher from 2 to 3 dpi. Haptoglobin (Hp) was significantly elevated from 3 dpi to the end of study, while pig major acute phase protein (Pig-MAP) from 3 to 7 dpi. The concentrations of CRP, Hp and SAA significantly increased before specific antibodies were detected. Positive correlations were found between serum concentration of Hp and SAA and lung scores, and between clinical score and concentrations of Pig-MAP and SAA. Conclusions The results of current study confirmed that monitoring of APP may revealed ongoing infection, and in this way may be useful in selecting clinically healthy pigs (i.e. before integration into an uninfected herd). Present results corroborated our previous findings that SAA could be a potentially useful indicator in experimental infection studies (e.g. vaccine efficiency investigations) or as a marker for disease severity, because of correlation observed between its concentration in serum and disease severity (lung scores, clinical scores). PMID:23332090

2013-01-01

370

Maternal Serum Disintegrin and Metalloprotease Protein-12 in Early Pregnancy as a Potential Marker of Adverse Pregnancy Outcomes  

PubMed Central

Objectives The aim of this study was to determine whether the concentration of disintegrin and metalloprotease protein12 (ADAM12) in first trimester maternal serum can be used as a marker for first-trimester complete spontaneous abortions, missed abortions, ectopic pregnancies and hydatidiform moles. Methods The maternal serum concentrations of ADAM12 were measured in the range of 5–9+6 weeks of gestation using an automated AutoDelfia immunoassay platform in 9 cases of complete spontaneous abortion, 27 cases of missed abortions, 56 cases of ectopic pregnancies, 12 cases of hydatidiform moles, and 100 controls. Logistic regression analysis was used to determine significant factors for predicting adverse pregnancy outcomes in early pregnancy. Screening performance was assessed using receiver operating characteristic curves. Results Two hundred and four women were enrolled in the study. In the control group, the level of ADAM12 increased with gestational age. The median ADAM12 levels in the spontaneous abortion (0.430 MoM), ectopic pregnancy (0.460 MoM) and hydatidiform mole (0.037 MoM) groups were lower than that in the control group, while the median ADAM12 level in the missed abortion group (1.062 MoM) was not significant from the controls (1.002 MoM). Logistic regression analysis demonstrated that the level of ADAM12 in maternal serum facilitated the detection of ectopic pregnancies (OR?=?0.909; 95% CI?=?0.841?0.982) and complete spontaneous abortion (OR?=?0.863; 95% CI?=?0.787?0.946). Conclusions In complete spontaneous abortion and ectopic pregnancy, ADAM12 maintained at low levels in early pregnancies, and there were significant differences compared to normal pregnancies. ADAM12 is a promising marker for the diagnosis of complete spontaneous abortion and ectopic pregnancy in symptomatic women, and under certain conditions, ADAM12 can diagnose ectopic pregnancy and spontaneous abortion before an ultrasonographic detection of the conditions. PMID:24830297

Yang, Jiexia; Wu, Jing; Guo, Fangfang; Wang, Dongmei; Chen, Keyi; Li, Jie; Du, Li; Yin, Aihua

2014-01-01

371

Constraint-based Local Move Definitions for Lattice Protein Models Including Side Chains  

E-print Network

The simulation of a protein's folding process is often done via stochastic local search, which requires a procedure to apply structural changes onto a given conformation. Here, we introduce a constraint-based approach to enumerate lattice protein structures according to k-local moves in arbitrary lattices. Our declarative description is much more flexible for extensions than standard operational formulations. It enables a generic calculation of k-local neighbors in backbone-only and side chain models. We exemplify the procedure using a simple hierarchical folding scheme.

Mann, Martin; Steinhöfel, Kathleen; Backofen, Rolf

2009-01-01

372

Multidimensional separation of tryptic peptides from human serum proteins using reversed-phase, strong cation exchange, weak anion exchange, and fused-core fluorinated stationary phases.  

PubMed

Proteome profiling of crude serum is a challenging task due to the wide dynamic range of protein concentrations and the presence of high-abundance proteins, which cover >90% of the total protein mass in serum. Peptide fractionation on strong cation exchange, weak anion exchange in the electrostatic repulsion hydrophilic interaction chromatography (ERLIC) mode, RP C18 at pH 2.5 (low pH), fused-core fluorinated at pH 2.5, and RP C18 at pH 9.7 (high pH) stationary phases resulted in two to three times more identified proteins and three to four times more identified peptides in comparison with 1D nanoChip-LC-MS/MS quadrupole TOF analysis (45 proteins, 185 peptides). The largest number of peptides and proteins was identified after prefractionation in the ERLIC mode due to the more uniform distribution of peptides among the collected fractions and on the RP column at high pH due to the high efficiency of RP separations and the complementary selectivity of both techniques to low-pH RP chromatography. A 3D separation scheme combining ERLIC, high-pH RP, and low-pH nanoChip-LC-MS/MS for crude serum proteome profiling resulted in the identification of 208 proteins and 1088 peptides with the lowest reported concentration of 11 ng/mL for heat shock protein 74. PMID:24039020

Boichenko, Alexander P; Govorukhina, Natalia; van der Zee, Ate G J; Bischoff, Rainer

2013-11-01

373

Detection of the Inflammation Biomarker C-Reactive Protein in Serum Samples: Towards an Optimal Biosensor Formula  

PubMed Central

The development of an electrochemical immunosensor for the biomarker, C-reactive protein (CRP), is reported in this work. CRP has been used to assess inflammation and is also used in a multi-biomarker system as a predictive biomarker for cardiovascular disease risk. A gold-based working electrode sensor was developed, and the types of electrode printing inks and ink curing techniques were then optimized. The electrodes with the best performance parameters were then employed for the construction of an immunosensor for CRP by immobilizing anti-human CRP antibody on the working electrode surface. A sandwich enzyme-linked immunosorbent assay (ELISA) was then constructed after sample addition by using anti-human CRP antibody labelled with horseradish peroxidase (HRP). The signal was generated by the addition of a mediator/substrate system comprised of 3,3,5',5'-Tetramethylbenzidine dihydrochloride (TMB) and hydrogen peroxide (H2O2). Measurements were conducted using chronoamperometry at ?200 mV against an integrated Ag/AgCl reference electrode. A CRP limit of detection (LOD) of 2.2 ng·mL?1 was achieved in spiked serum samples, and performance agreement was obtained with reference to a commercial ELISA kit. The developed CRP immunosensor was able to detect a diagnostically relevant range of the biomarker in serum without the need for signal amplification using nanoparticles, paving the way for future development on a cardiac panel electrochemical point-of-care diagnostic device. PMID:25587427

Fakanya, Wellington M.; Tothill, Ibtisam E.

2014-01-01

374

A serum “sweet-doughnut” protein facilitates fibrosis evaluation and therapy assessment in patients with viral hepatitis  

PubMed Central

Although liver fibrosis reflects disease severity in chronic hepatitis patients, there has been no simple and accurate system to evaluate the therapeutic effect based on fibrosis. We developed a glycan-based immunoassay, FastLec-Hepa, to fill this unmet need. FastLec-Hepa automatically detects unique fibrosis-related glyco-alteration in serum hyperglycosylated Mac-2 binding protein within 20?min. The serum FastLec-Hepa counts increased with advancing fibrosis and illustrated significant differences in medians between all fibrosis stages. FastLec-Hepa is sufficiently sensitive and quantitative to evaluate the effects of PEG-interferon-?/ribavirin therapy in a short post-therapeutic interval. The obtained fibrosis progression is equivalent to -0.30 stages/year in patients with sustained virological response, and 0.01 stages/year in relapse/nonresponders. Furthermore, long-term follow-up of the severely affected patients found hepatocellular carcinoma developed in patients after therapy whose FastLec-Hepa counts remained above a designated cutoff value. FastLec-Hepa is the only assay currently available for clinically beneficial therapy evaluation through quantitation of disease severity. PMID:23323209

Kuno, Atsushi; Ikehara, Yuzuru; Tanaka, Yasuhito; Ito, Kiyoaki; Matsuda, Atsushi; Sekiya, Satoru; Hige, Shuhei; Sakamoto, Michiie; Kage, Masayoshi; Mizokami, Masashi; Narimatsu, Hisashi

2013-01-01

375

Interaction of the anticancer gallium(III) complexes of 8-hydroxyquinoline and maltol with human serum proteins.  

PubMed

Tris(8-quinolinolato)gallium(III) (KP46) and tris(maltolato)gallium(III) (GaM) are promising orally active antitumor metallodrugs currently undergoing clinical trials. Their interaction with human serum albumin (HSA) and transferrin (Tf) was studied in detail in aqueous solution by the combination of various methods such as spectrofluorometry, UV-vis spectrophotometry, (1)H and saturation transfer difference NMR spectroscopy, and ultrafiltration-UV-vis spectrophotometry. Binding data were evaluated quantitatively. Tf was found to replace the original ligand much less efficiently in KP46 than in GaM, whereas a significant noncovalent binding of KP46 with HSA (log K' = 4.04) retaining the coordination environment around gallium(III) was found. The interaction between HSA and KP46 was also confirmed by protein-complex modeling calculations. On the basis of the conditional stability constants, the distribution of gallium(III) in serum was computed and compared for these metallodrugs under physiological conditions, and revealed the prominent role of HSA in the case of KP46 and that of Tf for GaM. PMID:25398250

Enyedy, Éva A; Dömötör, Orsolya; Bali, Krisztina; Hetényi, Anasztázia; Tuccinardi, Tiziano; Keppler, Bernhard K

2015-01-01

376

Detection of the inflammation biomarker C-reactive protein in serum samples: towards an optimal biosensor formula.  

PubMed

The development of an electrochemical immunosensor for the biomarker, C-reactive protein (CRP), is reported in this work. CRP has been used to assess inflammation and is also used in a multi-biomarker system as a predictive biomarker for cardiovascular disease risk. A gold-based working electrode sensor was developed, and the types of electrode printing inks and ink curing techniques were then optimized. The electrodes with the best performance parameters were then employed for the construction of an immunosensor for CRP by immobilizing anti-human CRP antibody on the working electrode surface. A sandwich enzyme-linked immunosorbent assay (ELISA) was then constructed after sample addition by using anti-human CRP antibody labelled with horseradish peroxidase (HRP). The signal was generated by the addition of a mediator/substrate system comprised of 3,3,5',5'-Tetramethylbenzidine dihydrochloride (TMB) and hydrogen peroxide (H2O2). Measurements were conducted using chronoamperometry at -200 mV against an integrated Ag/AgCl reference electrode. A CRP limit of detection (LOD) of 2.2 ng·mL(-1) was achieved in spiked serum samples, and performance agreement was obtained with reference to a commercial ELISA kit. The developed CRP immunosensor was able to detect a diagnostically relevant range of the biomarker in serum without the need for signal amplification using nanoparticles, paving the way for future development on a cardiac panel electrochemical point-of-care diagnostic device. PMID:25587427

Fakanya, Wellington M; Tothill, Ibtisam E

2014-12-01

377

Heterogeneity of human serum amyloid A protein. Five different variants from one individual demonstrated by cDNA sequence analysis.  

PubMed Central

Serum amyloid A (SAA), a chemically polymorphic protein, is the most sensitive marker protein of the acute phase and the precursor of reactive amyloidosis, which is characterized by deposits of amyloid A protein (AA). We investigated the variability of the SAA gene family in one individual by sequencing 11 SAA-specific clones from an acute-phase-liver cDNA library. At least five different SAA variants were deduced from six different cDNAs. The 3' untranslated gene segments fall into two groups, based on nucleotide sequence and variability in length. Various nucleotide and amino acid substitutions were found predominantly in the 3' portion. Some of these substitutions are unique and increase the number of SAA variants in one individual to at least five. Moreover, genomic DNA of four individuals was examined by analysis of restriction-fragment length polymorphism. Besides two conserved strongly labelled bands, additional polymorphic bands were observed, indicating isotypic and/or allotypic SAA variations. Finally, three different mRNA species were detected by Northern-blot analysis, a finding that might be of relevance for the stability of SAA transcripts. Images Fig. 5. Fig. 6. PMID:1971508

Steinkasserer, A; Weiss, E H; Schwaeble, W; Linke, R P

1990-01-01

378

Analyses of cardiac blood cells and serum proteins with regard to cause of death in forensic autopsy cases.  

PubMed

To investigate hematological and serum protein profiles of cadaveric heart blood with regard to the cause of death, serial forensic autopsy cases (n=308, >18 years of age, within 48 h postmortem) were examined. Red blood cells (Rbc), hemoglobin (Hb), platelets (Plt), white blood cells (Wbc), total protein (TP) and albumin (Alb) were examined in bilateral cardiac blood. Blood cell counts, collected after turning the bodies at autopsy, approximated to the clinical values. Postmortem changes were not significant for these markers. In non-head blunt injury cases, Rbc counts, Hb, TP and Alb levels in bilateral cardiac blood were lower in subacute deaths (survival time, 1-12 h) than in acute deaths (survival time <1 h). Wbc counts of left cardiac blood were significantly higher for non-head injury than for head injury in subacute deaths. In fire fatality cases, Plt count was markedly higher with an automated hematology analyzer than by using a blood smear test, suggesting Rbc fragmentation caused by deep burns, while increases in Wbc count and decreases in Alb levels were seen for subacute deaths. For asphyxiation, Rbc count, Hb, TP and Alb levels in bilateral cardiac blood were higher than other groups, and TP and Alb levels in the right cardiac blood were higher for hanging than for strangulation. These findings suggest that analyses of blood cells and proteins are useful for investigating the cause of death. PMID:19261512

Quan, Li; Ishikawa, Takaki; Michiue, Tomomi; Li, Dong-Ri; Zhao, Dong; Yoshida, Chiemi; Chen, Jian-Hua; Komatsu, Ayumi; Azuma, Yoko; Sakoda, Shigeki; Zhu, Bao-Li; Maeda, Hitoshi

2009-04-01

379

Air filter devices including nonwoven meshes of electrospun recombinant spider silk proteins.  

PubMed

Based on the natural sequence of Araneus diadematus Fibroin 4 (ADF4), the recombinant spider silk protein eADF4(C16) has been engineered. This highly repetitive protein has a molecular weight of 48kDa and is soluble in different solvents (hexafluoroisopropanol (HFIP), formic acid and aqueous buffers). eADF4(C16) provides a high potential for various technical applications when processed into morphologies such as films, capsules, particles, hydrogels, coatings, fibers and nonwoven meshes. Due to their chemical stability and controlled morphology, the latter can be used to improve filter materials. In this protocol, we present a procedure to enhance the efficiency of different air filter devices, by deposition of nonwoven meshes of electrospun recombinant spider silk proteins. Electrospinning of eADF4(C16) dissolved in HFIP results in smooth fibers. Variation of the protein concentration (5-25% w/v) results in different fiber diameters (80-1,100 nm) and thus pore sizes of the nonwoven mesh. Post-treatment of eADF4(C16) electrospun from HFIP is necessary since the protein displays a predominantly ?-helical secondary structure in freshly spun fibers, and therefore the fibers are water soluble. Subsequent treatment with ethanol vapor induces formation of water resistant, stable ?-sheet structures, preserving the morphology of the silk fibers and meshes. Secondary structure analysis was performed using Fourier transform infrared spectroscopy (FTIR) and subsequent Fourier self-deconvolution (FSD). The primary goal was to improve the filter efficiency of existing filter substrates by adding silk nonwoven layers on top. To evaluate the influence of electrospinning duration and thus nonwoven layer thickness on the filter efficiency, we performed air permeability tests in combination with particle deposition measurements. The experiments were carried out according to standard protocols. PMID:23685883

Lang, Gregor; Jokisch, Stephan; Scheibel, Thomas

2013-01-01

380

Air Filter Devices Including Nonwoven Meshes of Electrospun Recombinant Spider Silk Proteins  

PubMed Central

Based on the natural sequence of Araneus diadematus Fibroin 4 (ADF4), the recombinant spider silk protein eADF4(C16) has been engineered. This highly repetitive protein has a molecular weight of 48kDa and is soluble in different solvents (hexafluoroisopropanol (HFIP), formic acid and aqueous buffers). eADF4(C16) provides a high potential for various technical applications when processed into morphologies such as films, capsules, particles, hydrogels, coatings, fibers and nonwoven meshes. Due to their chemical stability and controlled morphology, the latter can be used to improve filter materials. In this protocol, we present a procedure to enhance the efficiency of different air filter devices, by deposition of nonwoven meshes of electrospun recombinant spider silk proteins. Electrospinning of eADF4(C16) dissolved in HFIP results in smooth fibers. Variation of the protein concentration (5-25% w/v) results in different fiber diameters (80-1,100 nm) and thus pore sizes of the nonwoven mesh. Post-treatment of eADF4(C16) electrospun from HFIP is necessary since the protein displays a predominantly ?-helical secondary structure in freshly spun fibers, and therefore the fibers are water soluble. Subsequent treatment with ethanol vapor induces formation of water resistant, stable ?-sheet structures, preserving the morphology of the silk fibers and meshes. Secondary structure analysis was performed using Fourier transform infrared spectroscopy (FTIR) and subsequent Fourier self-deconvolution (FSD). The primary goal was to improve the filter efficiency of existing filter substrates by adding silk nonwoven layers on top. To evaluate the influence of electrospinning duration and thus nonwoven layer thickness on the filter efficiency, we performed air permeability tests in combination with particle deposition measurements. The experiments were carried out according to standard protocols. PMID:23685883

Lang, Gregor; Jokisch, Stephan; Scheibel, Thomas

2013-01-01

381

Inhibition of the binding of low density lipoproteins to liver membrane receptors by rat serum phosphorylcholine binding protein.  

PubMed

Rat serum phosphorylcholine binding protein (PCBP) is characterized by its Ca2+ dependent property to bind phosphorylcholine ligand. PCBP immobilized on sepharose has been shown to selectively bind human plasma apo B and E containing lipoproteins. The present report describes an inhibitory effect of PCBP on the binding of human 125I-LDL to LDL receptors on estradiol treated rat liver membranes. Pre-incubation of liver membranes with PCBP did not affect the binding of 125I-LDL to the membranes. Gel filtration analysis of the incubation products from the LDL-receptor assay showed a concentration dependent binding of 125I-PCBP to LDL. The inhibitory effect of PCBP is likely due to the formation of LDL-PCBP complex and not due to the binding of PCBP to the LDL receptor site. PMID:3800992

Saxena, U; Nagpurkar, A; Mookerjea, S

1986-11-26

382

Fluorescent probing of protein bovine serum albumin stability and denaturation using polarity sensitive spectral response of a charge transfer probe.  

PubMed

The polarity sensitive photo-induced intra-molecular charge transfer (ICT) fluorescence probe (E)-3-(4-methylamino-phenyl)-acrylic acid ethyl ester (MAPAEE) has been used to study the model protein Bovine Serum Albumin (BSA) in its native and thermal and urea induced denatured states. The interaction between BSA and the regular surfactant Sodium Dodecyl Sulphate (SDS) as well as the biologically relevant steroid-based amphiphile Sodium Deoxycholate (NaDC) has also been very keenly followed using this ICT probe. The variation of micellar properties of both SDS and NaDC with increasing ionic strengths and in presence of the chaotrope urea has also been well documemted by the same probe. Steady-state spectroscopy, FRET, and fluorescence anisotropy measurements have been used to gain better insight into these processes and the molecule MAPAEE to be a full-bodied fluorescent probe for studying such intricate biological systems, their properties and interactions. PMID:20922468

Ghosh, Shalini; Jana, Sankar; Nath, Debnarayan; Guchhait, Nikhil

2011-01-01

383

High prevalence of serum antibodies reacting with simian virus 40 capsid protein mimotopes in patients affected by malignant pleural mesothelioma.  

PubMed

Human malignant pleural mesothelioma (MPM) is considered a rare tumor, but recent estimations indicate that one-quarter million people will die of this neoplasm in Europe in the next three decades. The mineral asbestos is considered the main causative agent of this neoplasm. MPM is largely unresponsive to conventional chemotherapy/radiotherapy. In addition to asbestos exposure, genetic predisposition to asbestos carcinogenesis and to simian virus (SV)40 infection has also been suggested. SV40 is a DNA tumor virus found in some studies to be associated at high prevalence with MPM. SV40 sequences have also been detected, although at a lower prevalence than in MPM, in blood specimens from healthy donors. However, some studies have failed to reveal SV40 footprints in MPM and its association with this neoplasm. These conflicting results indicate the need for further investigations with new approaches. We report on the presence of antibodies in serum samples from patients affected by MPM that specifically react with two different SV40 mimotopes. The two SV40 peptides used in indirect ELISAs correspond to viral capsid proteins. ELISA with the two SV40 mimotopes gave overlapping results. Our data indicate that in serum samples from MPM-affected patients (n = 97), the prevalence of antibodies against SV40 viral capsid protein antigens is significantly higher (26%, P = 0.043) than in the control group (15%) represented by healthy subjects (n = 168) with the same median age (66 y) and sex. Our results suggest that SV40 is associated with a subset of MPM and circulates in humans. PMID:23071320

Mazzoni, Elisa; Corallini, Alfredo; Cristaudo, Alfonso; Taronna, Angelo; Tassi, Gianfranco; Manfrini, Marco; Comar, Manola; Bovenzi, Massimo; Guaschino, Roberto; Vaniglia, Francesca; Magnani, Corrado; Casali, Ferruccio; Rezza, Giovanni; Barbanti-Brodano, Giuseppe; Martini, Fernanda; Tognon, Mauro G

2012-10-30

384

Human Cornea Proteome: Identification and Quantitation of the Proteins of the Three Main Layers Including Epithelium, Stroma, and Endothelium  

PubMed Central

Diseases of the cornea are common and refer to conditions like infections, injuries and genetic defects. Morphologically, many corneal diseases affect only certain layers of the cornea and separate analysis of the individual layers is therefore of interest to explore the basic molecular mechanisms involved in corneal health and disease. In this study, the three main layers including, the epithelium, stroma and endothelium of healthy human corneas were isolated. Prior to analysis by LC–MS/MS the proteins from the different layers were either (i) separated by SDS-PAGE followed by in-gel trypsinization, (ii) in-solution digested without prior protein separation or, (iii) in-solution digested followed by cation exchange chromatography. A total of 3250 unique Swiss-Prot annotated proteins were identified in human corneas, 2737 in the epithelium, 1679 in the stroma, and 880 in the endothelial layer. Of these, 1787 proteins have not previously been identified in the human cornea by mass spectrometry. In total, 771 proteins were quantified, 157 based on in-solution digestion and 770 based on SDS-PAGE separation followed by in-gel digestion of excised gel pieces. Protein analysis showed that many of the identified proteins are plasma proteins involved in defense responses. PMID:22698189

2012-01-01

385

Antibody responses against non-structural protein 3 of bovine viral diarrhoea virus in milk and serum samples from animals immunised with an inactivated vaccine.  

PubMed

Antibodies against non-structural protein 3 (NS3, p80) of bovine viral diarrhoea virus (BVDV) were determined in milk from cows vaccinated with an inactivated BVDV vaccine and compared to serum antibody levels. Animals in one herd were vaccinated with an inactivated BVDV vaccine according to the standard protocol and animals from a second herd with an intensive schedule. Serum and milk samples were tested for BVDV NS3 antibodies using five commercial ELISAs. With a few exceptions, vaccination according to the standard schedule did not induce BVDV NS3-specific antibodies in serum or milk. However, after vaccination according to the intensive schedule, anti-NS3 antibodies were detected for a short time in serum and, to a lesser extent, in milk. Bulk milk was a suitable substrate for BVDV monitoring of herds vaccinated with the inactivated BVD vaccine. PMID:21482158

Alvarez, Marcelino; Donate, Jorge; Makoschey, Birgit

2012-03-01

386

Evidence for IgG autoantibodies to galectin-3, a beta-galactoside-binding lectin (Mac-2, epsilon binding protein, or carbohydrate binding protein 35) in human serum.  

PubMed

Galectin-3 is a beta-galactoside-binding animal lectin formerly called epsilon protein, Mac-2, carbohydrate binding protein 35, CBH 30, L-29, or L34. The possible occurrence of autoantibodies to galectin-3 was investigated because crosslinking of galectins bound to IgE or Fc epsilon RI might produce mediator release from mast cells or basophils. Unexpectedly, a control serum from an individual free of current allergic symptoms was found to have a significantly elevated level of IgG anti-galectin-3 by ELISA employing galectin-3-coated wells incubated with test serum followed by HRPO-conjugated goat anti-human IgG. The reaction was not inhibitable by lactose, suggesting that it is not a result of binding of IgG by galectin-3 through lectin-carbohydrate interactions. The antibody activity was specifically adsorbed by galectin-3 and protein A-conjugated Sepharose and was associated primarily with subclass IgG1. The presence of the antibodies was confirmed by immunoblotting showing binding of IgG to the 30-kD galectin-3 band. The relevant epitopes were in the galectin-3 N-terminal domain. The propositus was subsequently found to have adenocarcinoma of the colon, and titers of IgG anti-galectin-3 were found to be sharply elevated after hemicolectomy. Similar antibody titers have not been found in family members, but small numbers of normal persons and patients with malignant neoplasms have