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1

Protein electrophoresis - serum  

MedlinePLUS

This lab test measures the types of protein in the fluid (serum) part of a blood sample. Other electrophoresis tests that measure proteins in the serum include: Immunoelectrophoresis Immunofixation Globulin electrophoresis

2

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

3

Immunohistochemical demonstration of serum proteins in human cerebral gliomas  

Microsoft Academic Search

The leakage of different serum proteins, including immunoglobulins, into human cerebral gliomas was studied by use of the unlabeled peroxidase-antiperoxidase (PAP) method on cryostat and paraffin sections. Our series of 50 tumour biopsies included 21 isomorphic astrocytomas and oligodendrogliomas (grade II), 19 anaplastic astrocytomas and oligodendrogliomas (grade III), and 10 glioblastomas (grade IV). The immunohistochemical staining of the serum proteins

R. J. Seitz; W. Wechsler

1987-01-01

4

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.

5

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

6

Binding of aluminum to human serum transferrin, human serum albumin and rat serum proteins  

SciTech Connect

Human serum transferrin (HSTF), human serum albumin (HSA) and rat serum were compared for their interaction with AlCl[sub 3], in a Tris-HCl buffer solutions. The AlCl[sub 3] was tested in series of concentrations in the range of 50 [mu]M up to 500 [mu]M. HSTF, HSA and their 1:1 mixture and rat serum were incubated at 37 C with series of AlCl[sub 3] concentrations. The protein profile of the incubated solutions were compared to control using SDS-PAGE and FPLC tests. The results indicated that HSTF was more specifically responsive to AlCl[sub 3] showing a characteristic increase in it UV absorption, peak and area dimensions. Simultaneously, HSA was less affected, but it showed a significant shift with an increase in molecular weight accompanied with a change in its profile. The respective bands of transferrin and albumin in rat serum behaved similarly.

El-Sebae, A.K.H.; Zeid, M.M.A.; Abdel-Rahman, F.H.; Saleh, M.A. (Texas Southern Univ., Houston, TX (United States). Environmental Chemistry and Toxicology Lab.)

1994-01-01

7

Quantitating metabolites in protein precipitated serum using NMR spectroscopy.  

PubMed

Quantitative NMR-based metabolite profiling is challenged by the deleterious effects of abundant proteins in the intact blood plasma/serum, which underscores the need for alternative approaches. Protein removal by ultrafiltration using low molecular weight cutoff filters thus represents an important step. However, protein precipitation, an alternative and simple approach for protein removal, lacks detailed quantitative assessment for use in NMR based metabolomics. In this study, we have comprehensively evaluated the performance of protein precipitation using methanol, acetonitrile, perchloric acid, and trichloroacetic acid and ultrafiltration approaches using 1D and 2D NMR, based on the identification and absolute quantitation of 44 human blood metabolites, including a few identified for the first time in the NMR spectra of human serum. We also investigated the use of a "smart isotope tag," (15)N-cholamine for further resolution enhancement, which resulted in the detection of a number of additional metabolites. (1)H NMR of both protein precipitated and ultrafiltered serum detected all 44 metabolites with comparable reproducibility (average CV, 3.7% for precipitation; 3.6% for filtration). However, nearly half of the quantified metabolites in ultrafiltered serum exhibited 10-74% lower concentrations; specifically, tryptophan, benzoate, and 2-oxoisocaproate showed much lower concentrations compared to protein precipitated serum. These results indicate that protein precipitation using methanol offers a reliable approach for routine NMR-based metabolomics of human blood serum/plasma and should be considered as an alternative to ultrafiltration. Importantly, protein precipitation, which is commonly used by mass spectrometry (MS), promises avenues for direct comparison and correlation of metabolite data obtained from the two analytical platforms to exploit their combined strength in the metabolomics of blood. PMID:24796490

Nagana Gowda, G A; Raftery, Daniel

2014-06-01

8

Production of recombinant protein C in serum-containing and serum-free perfusion culture  

Microsoft Academic Search

For the development of a perfusion culture producing recombinant human protein C, the effects of fetal calf serum and growth factors on cell growth and recombinant protein production were investigated. Although the growth of recombinant cells was stimulated by serum in a dose-dependent manner, a lower concentration of serum (2%) could support both synthesis and post-translational modification of protein C

Takeyuki Sugiura; Hiromi B. Maruyama

1991-01-01

9

Serum protein electrophoresis in 147 dogs  

Microsoft Academic Search

Reference intervals for serum protein electrophoresis (SPE) were created from a group of 75 clinically healthy dogs and compared with SPE results obtained from clinical cases presented to the University of Bristol over an eight-and-a-half-year period. A total of 147 dogs, in which SPE had been performed, had complete case records available and thus met the inclusion criteria. Signalment and

S. W. Tappin; S. S. Taylor; S. Tasker; S. J. Dodkin; K. Papasouliotis; K. F. Murphy

2011-01-01

10

Production of recombinant protein C in serum-containing and serum-free perfusion culture.  

PubMed

For the development of a perfusion culture producing recombinant human protein C, the effects of fetal calf serum and growth factors on cell growth and recombinant protein production were investigated. Although the growth of recombinant cells was stimulated by serum in a dose-dependent manner, a lower concentration of serum (2%) could support both synthesis and post-translational modification of protein C as efficiently as 10% serum. Among the growth factors tested, transferrin enhanced protein C production to the level comparable with 10% serum, while insulin was effective in maintaining cellular metabolism. Based on these results, a perfusion culture for a scale-up production of recombinant protein C was done using an Opticell culture system. A good productivity of the recombinant protein was obtained in low serum or serum-free medium for more than one month. PMID:1368118

Sugiura, T; Maruyama, H B

1991-11-01

11

Influence of a Diurnal Oxygen Pulse on Fish Serum Proteins  

Microsoft Academic Search

Changes in the serum protein, as shown by electrophoretic analyses, appear to be correlated with changes in the environment of the organism. These changes may be associated with aquatic pollution.A diurnal oxygen pulse of 3 ppm for 8 hours per day for 9 days produced a significant stress pattern in the serum protein fractions of bluegills and largemouth bass, but

Gerald R. Bouck; Robert C. Ball

1965-01-01

12

Effects of serum, its protein and lipid extracts, and commercial serum proteins and lipid on the isolated frog heart.  

PubMed

This study investigates the inotropic effects of serum, its protein and lipid extracts, and commercial serum proteins and lipid on the isolated, spontaneously-beating heart and superfused, hypodynamic ventricle of the frog. Serum taken from either man, horse, calf, frog, or rabbit evoked marked positive inotropic responses which were unaffected by cholinergic, serotonergic, and adrenergic receptor antagonists. Dialysed serum (dialisand) and void volume fractions from Sephadex G200-120 columns corresponding to large molecular weight constituents evoked marked positive inotropic responses. When serum was separated into fractions containing either proteins or lipids/lipoproteins by high-density ultracentrifugation or activated charcoal, both extracts evoked marked positive inotropic responses. Commercial serum globulins and serum containing a high proportion of immunoglobulins elicited large increases in contractile force, whereas serum albumin evoked a negative inotropic effect. Serum which was either boiled and/or treated with chymotrypsin to denature proteins also caused a marked increase in isometric twitch tension in the frog heart. Similar inotropic response was obtained with fractions of boiled serum eluted on columns of Sephadex G200-120. These fractions corresponded to molecular weight in the region of 60-70 kDa. However, the inotropic effect of boiled serum was abolished following pretreatment with lipase. Superfusion of frog hearts with commercial cardiolipin resulted in marked dose-dependent increases in contractile force. The results demonstrate the presence of at least two large molecular weight cardioactive principles in serum. These substances are comparable in size to constituents of serum proteins (e.g., globulins and immunoglobulins) and serum lipids/lipoproteins (e.g., cardiolipin) and may serve as physiological regulators of cardiac function. PMID:1939740

Singh, J; Hutton, T; Hussain, M; Waring, J J

1991-01-01

13

Micromethod for the Rapid Determination of Serum Protein-Bound Iodine and Total Serum Iodine  

Microsoft Academic Search

A modification is presentedof the chloric acid method for the rapid determination of serum protein-bound iodine and total serum iodine requiring less than 0.5 ml. serum for duplicate determinations. The time required for the entire procedure is lessthan two hours. Byadjusting the amountsof reagents used and by varying the time of digestion, the need for constant and continuoussupervisionis eliminated. 'IE1HEDETERMINATION

Eli Gardiner; Arthur Bums

14

Variations in protein binding of drugs in plasma and serum.  

PubMed

The free fractions of five drugs were determined in plasma treated with EDTA and in serum. The free fractions of phenytoin, sodium valproate, and phenobarbital in serum were significantly higher than in plasma (P less than 0.05). In contrast, the free fractions of carbamazepine and theophylline were significantly lower in serum than in plasma. Although we observed minor differences in the protein patterns obtained with two-dimensional electrophoresis, these did not have an important influence on protein binding. No significant differences were observed between plasma and serum in the physiological data, except for pH. Binding of these drugs by protein was shown to be pH-dependent. We conclude that serum may be better than plasma for determination of the free fraction, because the free fraction in plasma is affected by anticoagulants such as EDTA salts and heparin. PMID:2503268

Ohshima, T; Hasegawa, T; Johno, I; Kitazawa, S

1989-08-01

15

Spectroscopic imaging of serum proteins using quantum cascade lasers.  

PubMed

First measurements of biomedical imaging using quantum cascade lasers (QCL) are presented. We report spectroscopic imaging of serum proteins using QCLs as an example for monitoring surface biocontamination. We found that dry smears of human serum can be spectroscopically imaged, identified, and quantified with high sensitivity and specificity. The core parts of the imaging platform consist of optically multiplexing three QCLs and an uncooled microbolometer camera. We show imaging of human serum proteins at 6.1, 9.25, and 9.5 ?m QCLs with high sensitivity and specificity. The sensitivity limit of 3???g/cm² of the human serum spot was measured at an S/N=3.The specificity of human serum detection was measured at 99% probability at a threshold of 77???g/cm². We anticipate our imaging technique to be a starting point for more sophisticated biomolecular diagnostic applications. PMID:23515866

Mukherjee, Anadi; Bylund, Quentin; Prasanna, Manu; Margalit, Yotam; Tihan, Tarik

2013-03-01

16

Distinctive serum protein profiles involving abundant proteins in lung cancer patients based upon antibody microarray analysis  

Microsoft Academic Search

BACKGROUND: Cancer serum protein profiling by mass spectrometry has uncovered mass profiles that are potentially diagnostic for several common types of cancer. However, direct mass spectrometric profiling has a limited dynamic range and difficulties in providing the identification of the distinctive proteins. We hypothesized that distinctive profiles may result from the differential expression of relatively abundant serum proteins associated with

Wei-Min Gao; Rork Kuick; Randal P Orchekowski; David E Misek; Ji Qiu; Alissa K Greenberg; William N Rom; Dean E Brenner; Gilbert S Omenn; Brian B Haab; Samir M Hanash

2005-01-01

17

Simple Model of Membrane Proteins Including Solvent  

E-print Network

We report a numerical simulation for the phase diagram of a simple two dimensional model, similar to one proposed by Noro and Frenkel [J. Chem. Phys. \\textbf{114}, 2477 (2001)] for membrane proteins, but one that includes the role of the solvent. We first use Gibbs ensemble Monte Caro simulations to determine the phase behavior of particles interacting via a square-well potential in two dimensions for various values of the interaction range. A phenomenological model for the solute-solvent interactions is then studied to understand how the fluid-fluid coexistence curve is modified by solute-solvent interactions. It is shown that such a model can yield systems with liquid-liquid phase separation curves that have both upper and lower critical points, as well as closed loop phase diagrams, as is the case with the corresponding three dimensional model.

D. L. Pagan; A. Shiryayev; T. P. Connor; J. D. Gunton

2006-03-04

18

Detection of human serum proteins using Raman and SERS spectroscopy  

NASA Astrophysics Data System (ADS)

The use of normal Raman (NR) spectroscopy and surface enhanced Raman scattering (SERS) spectroscopy to analyze the biochemical information of human serum proteins and hence distinguish between normal and primary hepatic carcinoma (PHC) serum samples was investigated. The serum samples were obtained from patients who were clinically diagnosed with PHC (n=20) and healthy volunteers (n=20). All spectra were collected in the spectral range of 400-1800 cm-1 and analyzed through the multivariate statistical methods of principal component analysis (PCA). The results showed that both NR and SERS combined with PCA had good performance in distinguishing the human serum proteins between PHC patients and healthy volunteers with high sensitivity and specificity of 100%. And we can get more detail information of component and conformation of human serum proteins by considering NR and SERS spectrum. Our results support the concept again that serum protein Raman and SERS spectroscopy combined with PCA analysis both can become noninvasive and rapid diagnostic tools to detect the primary hepatic carcinoma.

Ruan, Qiuyong; Liao, Fadian; Lin, Juqiang; Liu, Nenrong; Lin, Jinyong; Zeng, Yongyi; Li, Ling; Huang, Zufang; Chen, Rong

2014-09-01

19

Protein Depletion for Plasma and Serum Proteomic Analysis  

E-print Network

, there is an increasing trend towards the use of blood plasma for proteomic profiling to ensure that important proteinsProtein Depletion for Plasma and Serum Proteomic Analysis TABLE OF CONTENTS INTRODUCTION · Why.g. PTM) as a result of disease or physiological state can potentially be used as diagnostic biomarkers

Lebendiker, Mario

20

Anion exchange fractionation of serum proteins versus albumin elimination.  

PubMed

Elimination of albumin, constituting more than 50% of total serum proteins, allows increased protein loads on immobilized pH gradient (IPG) gels and better visualization of low-abundance proteins; however, it may result in the loss of albumin-bound low-abundance proteins. In this study, we report the prefractionation of serum proteins by batch anion exchange chromatography into three fractions: one containing proteins with isoelectric points (pI values) higher than the pI of albumin, a second fraction containing proteins with pI values in the same range as the pI of albumin, and a third fraction containing proteins with pI values lower than the pI of albumin. This procedure uses common instrumentation, is carried out under denaturing conditions, and takes less than 30min. We also report the loss of a clinically established prostate cancer serum biomarker, prostate-specific antigen (PSA), after albumin is eliminated using two commercially available albumin elimination kits: one that uses Cibacron Blue F3GA, which achieves albumin depletion through dye-ligand binding, and one that uses specific albumin antibody. The loss of PSA secondary to albumin elimination exceeded that after batch anion exchange serum sample prefractionation. PMID:17618595

Sahab, Ziad J; Iczkowski, Kenneth A; Sang, Qing-Xiang Amy

2007-09-01

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

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 20 mg 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

23

Serum Resistin (FIZZ3) Protein Is Increased in Obese Humans  

Microsoft Academic Search

The role of resistin in obesity and insulin resistance in hu- mans is controversial. Therefore, resistin protein was quan- titated by ELISA in serum of 27 lean (13 women\\/14 men, body mass index (BMI) 21.7 0.4 kg\\/m2, age 33 2 yr) and 50 obese (37 women\\/13 men, BMI 49.8 1.5 kg\\/m2, age 47 1 yr) subjects. There was more serum

MIKAKO DEGAWA-YAMAUCHI; JASON E. BOVENKERK; BETH ELISA JULIAR; WILLIAM WATSON; KIMBERLY KERR; ROSEMARIE JONES; QIHONG ZHU; ROBERT V. CONSIDINE

2003-01-01

24

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

25

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

26

Serum protein profile in sickle cell disease.  

PubMed Central

The total protein, albumin, globulin, and immunoglobulin levels of sera from 96 children with homozygous sickle cell disease were studied. A comparison of the results with the levels found in a control group of normal children of the same age shows that the sicklers have higher total protein, globulin, and IgM levels. The amounts of albumin and IgA seen were almost the same in both groups. The IgG levels differed considerably, the sicklers having only about half the quantity seen in normal children. PMID:438342

Isichei, U P

1979-01-01

27

SERUM PROTEIN PROFILES IN THE SUCKLING AND NON SUCKLING PIGLET  

E-print Network

SERUM PROTEIN PROFILES IN THE SUCKLING AND NON SUCKLING PIGLET: THE IMPORTANCE OF COLOSTRUM J PORCELET NOUVEAU-NE : IMPOR- TANCE DU COLOSTRUM. ― Le profil des protéines sériques a été étudié, qui ont reçu le colostrum, un profil de type adulte est observé dans les deux heures qui suivent la

Paris-Sud XI, Université de

28

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

Microsoft Academic Search

In clinical and pharmaceutical proteomics, serum and plasma are frequently used for detection of early diagnostic biomarkers\\u000a for therapeutic targets. Although obtaining these body fluid samples is non-invasive and easy, they contain some abundant\\u000a proteins that mask other protein components present at low concentrations. The challenge in identifying serum biomarkers is\\u000a to remove the abundant proteins, uncovering and enriching at

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

2011-01-01

29

Relationship of the serum protein-bound iodine to rates of gain in beef cattle  

E-print Network

RELATIONSHIP OP THE SERUM PROTEIN-BOUND IODINE TO RATES OF GAIN IN BEEP CATTLE A Dissertation b7 DAVID KERSHAW STOKES, JR. Approved as to style and content by: January 1956 L IB R ARY R ft M COLLEGE OF TEXAS RELATIONSHIP OP THE SERUM... HERITABILITY AND REPEATABILITY OF THE SERUM PROTEIN- BOUND IODINE IN IMMATURE BEEF CATTLE...................... 13 THE METHOD OF MEASUREMENT OF SERUM PROTEIN- BOUND IODINE............................................ 13 REPEATABILITY OF THE SERUM PBI LEVEL...

Stokes, David Kershaw

1956-01-01

30

Depletion of multiple high-abundance proteins improves protein profiling capacities of human serum and plasma.  

PubMed

Systematic detection of low-abundance proteins in human blood that may be putative disease biomarkers is complicated by an extremely wide range of protein abundances. Hence, depletion of major proteins is one potential strategy for enhancing detection sensitivity in serum or plasma. This study compared a recently commercialized HPLC column containing antibodies to six of the most abundant blood proteins ("Top-6 depletion") with either older Cibacron blue/Protein A or G depletion methods or no depletion. In addition, a prototype spin column version of the HPLC column and an alternative prototype two antibody spin column were evaluated. The HPLC polyclonal antibody column and its spin column version are very promising methods for substantially simplifying human serum or plasma samples. These columns show the lowest nonspecific binding of the depletion methods tested. In contrast other affinity methods, particularly dye-based resins, yielded many proteins in the bound fractions in addition to the targeted proteins. Depletion of six abundant proteins removed about 85% of the total protein from human serum or plasma, and this enabled 10- to 20-fold higher amounts of depleted serum or plasma samples to be applied to 2-D gels or alternative protein profiling methods such as protein array pixelation. However, the number of new spots detected on 2-D gels was modest, and most newly visualized spots were minor forms of relatively abundant proteins. The inability to detect low-abundance proteins near expected 2-D staining limits was probably due to both the highly heterogeneous nature of most plasma or serum proteins and masking of many low-abundance proteins by the next series of most abundant proteins. Hence, non2-D methods such as protein array pixelation are more promising strategies for detecting lower abundance proteins after depleting the six abundant proteins. PMID:16052620

Echan, Lynn A; Tang, Hsin-Yao; Ali-Khan, Nadeem; Lee, KiBeom; Speicher, David W

2005-08-01

31

Discovery and Fine Mapping of Serum Protein Loci through Transethnic Meta-analysis  

PubMed Central

Many disorders are associated with altered serum protein concentrations, including malnutrition, cancer, and cardiovascular, kidney, and inflammatory diseases. Although these protein concentrations are highly heritable, relatively little is known about their underlying genetic determinants. Through transethnic meta-analysis of European-ancestry and Japanese genome-wide association studies, we identified six loci at genome-wide significance (p < 5 × 10?8) for serum albumin (HPN-SCN1B, GCKR-FNDC4, SERPINF2-WDR81, TNFRSF11A-ZCCHC2, FRMD5-WDR76, and RPS11-FCGRT, in up to 53,190 European-ancestry and 9,380 Japanese individuals) and three loci for total protein (TNFRS13B, 6q21.3, and ELL2, in up to 25,539 European-ancestry and 10,168 Japanese individuals). We observed little evidence of heterogeneity in allelic effects at these loci between groups of European and Japanese ancestry but obtained substantial improvements in the resolution of fine mapping of potential causal variants by leveraging transethnic differences in the distribution of linkage disequilibrium. We demonstrated a functional role for the most strongly associated serum albumin locus, HPN, for which Hpn knockout mice manifest low plasma albumin concentrations. Other loci associated with serum albumin harbor genes related to ribosome function, protein translation, and proteasomal degradation, whereas those associated with serum total protein include genes related to immune function. Our results highlight the advantages of transethnic meta-analysis for the discovery and fine mapping of complex trait loci and have provided initial insights into the underlying genetic architecture of serum protein concentrations and their association with human disease. PMID:23022100

Franceschini, Nora; van Rooij, Frank J.A.; Prins, Bram P.; Feitosa, Mary F.; Karakas, Mahir; Eckfeldt, John H.; Folsom, Aaron R.; Kopp, Jeffrey; Vaez, Ahmad; Andrews, Jeanette S.; Baumert, Jens; Boraska, Vesna; Broer, Linda; Hayward, Caroline; Ngwa, Julius S.; Okada, Yukinori; Polasek, Ozren; Westra, Harm-Jan; Wang, Ying A.; Del Greco M., Fabiola; Glazer, Nicole L.; Kapur, Karen; Kema, Ido P.; Lopez, Lorna M.; Schillert, Arne; Smith, Albert V.; Winkler, Cheryl A.; Zgaga, Lina; Bandinelli, Stefania; Bergmann, Sven; Boban, Mladen; Bochud, Murielle; Chen, Y.D.; Davies, Gail; Dehghan, Abbas; Ding, Jingzhong; Doering, Angela; Durda, J. Peter; Ferrucci, Luigi; Franco, Oscar H.; Franke, Lude; Gunjaca, Grog; Hofman, Albert; Hsu, Fang-Chi; Kolcic, Ivana; Kraja, Aldi; Kubo, Michiaki; Lackner, Karl J.; Launer, Lenore; Loehr, Laura R.; Li, Guo; Meisinger, Christa; Nakamura, Yusuke; Schwienbacher, Christine; Starr, John M.; Takahashi, Atsushi; Torlak, Vesela; Uitterlinden, André G.; Vitart, Veronique; Waldenberger, Melanie; Wild, Philipp S.; Kirin, Mirna; Zeller, Tanja; Zemunik, Tatijana; Zhang, Qunyuan; Ziegler, Andreas; Blankenberg, Stefan; Boerwinkle, Eric; Borecki, Ingrid B.; Campbell, Harry; Deary, Ian J.; Frayling, Timothy M.; Gieger, Christian; Harris, Tamara B.; Hicks, Andrew A.; Koenig, Wolfgang; O’Donnell, Christopher J.; Fox, Caroline S.; Pramstaller, Peter P.; Psaty, Bruce M.; Reiner, Alex P.; Rotter, Jerome I.; Rudan, Igor; Snieder, Harold; Tanaka, Toshihiro; van Duijn, Cornelia M.; Vollenweider, Peter; Waeber, Gerard; Wilson, James F.; Witteman, Jacqueline C.M.; Wolffenbuttel, Bruce H.R.; Wright, Alan F.; Wu, Qingyu; Liu, Yongmei; Jenny, Nancy S.; North, Kari E.; Felix, Janine F.; Alizadeh, Behrooz Z.; Cupples, L. Adrienne; Perry, John R.B.; Morris, Andrew P.

2012-01-01

32

Discovery and fine mapping of serum protein loci through transethnic meta-analysis.  

PubMed

Many disorders are associated with altered serum protein concentrations, including malnutrition, cancer, and cardiovascular, kidney, and inflammatory diseases. Although these protein concentrations are highly heritable, relatively little is known about their underlying genetic determinants. Through transethnic meta-analysis of European-ancestry and Japanese genome-wide association studies, we identified six loci at genome-wide significance (p < 5 × 10(-8)) for serum albumin (HPN-SCN1B, GCKR-FNDC4, SERPINF2-WDR81, TNFRSF11A-ZCCHC2, FRMD5-WDR76, and RPS11-FCGRT, in up to 53,190 European-ancestry and 9,380 Japanese individuals) and three loci for total protein (TNFRS13B, 6q21.3, and ELL2, in up to 25,539 European-ancestry and 10,168 Japanese individuals). We observed little evidence of heterogeneity in allelic effects at these loci between groups of European and Japanese ancestry but obtained substantial improvements in the resolution of fine mapping of potential causal variants by leveraging transethnic differences in the distribution of linkage disequilibrium. We demonstrated a functional role for the most strongly associated serum albumin locus, HPN, for which Hpn knockout mice manifest low plasma albumin concentrations. Other loci associated with serum albumin harbor genes related to ribosome function, protein translation, and proteasomal degradation, whereas those associated with serum total protein include genes related to immune function. Our results highlight the advantages of transethnic meta-analysis for the discovery and fine mapping of complex trait loci and have provided initial insights into the underlying genetic architecture of serum protein concentrations and their association with human disease. PMID:23022100

Franceschini, Nora; van Rooij, Frank J A; Prins, Bram P; Feitosa, Mary F; Karakas, Mahir; Eckfeldt, John H; Folsom, Aaron R; Kopp, Jeffrey; Vaez, Ahmad; Andrews, Jeanette S; Baumert, Jens; Boraska, Vesna; Broer, Linda; Hayward, Caroline; Ngwa, Julius S; Okada, Yukinori; Polasek, Ozren; Westra, Harm-Jan; Wang, Ying A; Del Greco M, Fabiola; Glazer, Nicole L; Kapur, Karen; Kema, Ido P; Lopez, Lorna M; Schillert, Arne; Smith, Albert V; Winkler, Cheryl A; Zgaga, Lina; Bandinelli, Stefania; Bergmann, Sven; Boban, Mladen; Bochud, Murielle; Chen, Y D; Davies, Gail; Dehghan, Abbas; Ding, Jingzhong; Doering, Angela; Durda, J Peter; Ferrucci, Luigi; Franco, Oscar H; Franke, Lude; Gunjaca, Grog; Hofman, Albert; Hsu, Fang-Chi; Kolcic, Ivana; Kraja, Aldi; Kubo, Michiaki; Lackner, Karl J; Launer, Lenore; Loehr, Laura R; Li, Guo; Meisinger, Christa; Nakamura, Yusuke; Schwienbacher, Christine; Starr, John M; Takahashi, Atsushi; Torlak, Vesela; Uitterlinden, André G; Vitart, Veronique; Waldenberger, Melanie; Wild, Philipp S; Kirin, Mirna; Zeller, Tanja; Zemunik, Tatijana; Zhang, Qunyuan; Ziegler, Andreas; Blankenberg, Stefan; Boerwinkle, Eric; Borecki, Ingrid B; Campbell, Harry; Deary, Ian J; Frayling, Timothy M; Gieger, Christian; Harris, Tamara B; Hicks, Andrew A; Koenig, Wolfgang; O' Donnell, Christopher J; Fox, Caroline S; Pramstaller, Peter P; Psaty, Bruce M; Reiner, Alex P; Rotter, Jerome I; Rudan, Igor; Snieder, Harold; Tanaka, Toshihiro; van Duijn, Cornelia M; Vollenweider, Peter; Waeber, Gerard; Wilson, James F; Witteman, Jacqueline C M; Wolffenbuttel, Bruce H R; Wright, Alan F; Wu, Qingyu; Liu, Yongmei; Jenny, Nancy S; North, Kari E; Felix, Janine F; Alizadeh, Behrooz Z; Cupples, L Adrienne; Perry, John R B; Morris, Andrew P

2012-10-01

33

Investigation of change of serum immunosuppressive acidic protein levels in gynecological tumors  

Microsoft Academic Search

pressive acidic protein ( IAP) levels to diagnose and follow up survey patients with the gynecological tumor. Methods : Serum IAP levels were determined by IAP2single radial immunodiffusion test in 235 patients with the gynecological tumor ,including 38 cases of benign tumor of ovary ,41 cases of malignant tumor of ovary ,66 cases of hysteromyoma ,34 carcinomas of uterine cervix

Zhang Weiyuan; Sun Bo; Chen Mo; Zhao Yanhui

2002-01-01

34

Serum Immune-Related Proteins are Differentially Expressed during Hibernation in the American Black Bear  

PubMed Central

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

35

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

36

Biotransformation of BMOV in the presence of blood serum proteins.  

PubMed

The interaction of the potent anti-diabetic agent bis(maltolato)oxidovanadium(IV) (BMOV) with some proteins of blood serum was studied by EPR spectroscopy, pH-potentiometry and DFT calculations. The formation of cis-VO(ma)(2)(hTf), cis-VO(ma)(2)(HSA) and cis-VO(ma)(2)(IgG), their role in the biotransformation in vivo and the mechanism of transport of BMOV in blood are discussed. PMID:22094980

Sanna, Daniele; Bíró, Linda; Buglyó, Péter; Micera, Giovanni; Garribba, Eugenio

2012-01-01

37

Serum proteins of Canada goose (Branta canadensis) subspecies  

USGS Publications Warehouse

Serum proteins from nine subspecies of Canada Geese (Brunta canadensis) were analyzed through the use of column and slab acrylamide electrophoresis. Variation was minimal within a subspecies, although all the subspecies were closely related. B. c. leucopareia appeared to be the most distinct subspecies, while maxima and moffitti were the most similar. Our preliminary findings suggest that the electrophoresis techniques are sensitive enough to identify some of the subspecies; however, baseline data from breeding ranges of all subspecies are required.

Morgan, R.P., II; Sulkin, S.T.; Henny, C.J.

1977-01-01

38

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

39

Detection of pancreatic cancer using serum protein profiling  

PubMed Central

Background Currently, no suitable biomarkers for the early detection of pancreatic cancer (PC) are available. Proteins present in the serum could reflect a state of the disease. In this study, these profiles as a diagnostic marker for PC were evaluated. Methods Serum samples were obtained from PC patients (n = 50 calibration set, n = 39 validation set) and healthy volunteers (n = 110 and n = 75 respectively) according to a uniform standardized collection and processing protocol. For peptide and protein isolation, automated solid-phase extraction (SPE) with Weak Cation Exchange (WCX) magnetic beads (MB) was performed using a 96-channel liquid handling platform. Protein profiles were obtained by mass spectrometry (MS) and evaluated by linear discriminant analysis with double cross-validation. Results A discriminating profile for PC has been identified, with a sensitivity of 78% and a specificity of 89% in the calibration set with an area under the curve (AUC) of 90%. These results were validated with a sensitivity of 74% and a specificity of 91% (AUC 90%). Conclusion Serum profiles of healthy controls and PC can be discrimated between. Further research is warranted to evaluate specificity and whether this biosignature can be used for early detection in a high risk population. PMID:23458426

Velstra, Berit; Bonsing, Bert A; Mertens, Bart J; Burgt, Yuri E M; Huijbers, Anouck; Vasen, Hans; Mesker, Wilma E; Deelder, André M; Tollenaar, Rob A E M

2013-01-01

40

Serum protein profile of Crohn's disease treated with infliximab.  

PubMed

The infliximab (IFX) has dramatically improved the treatment of Crohn's disease (CD). However, the need for predictive factors, indicative of patients' response to IFX, has yet to be met. In the current study, proteomics technologies were employed in order to monitor for differences in protein expression in a cohort of patients following IFX administration, aiming at identifying a panel of candidate protein biomarkers of CD, symptomatic of response to treatment. We enrolled 18 patients, who either had achieved clinical and serological remission (Rm, n=6), or response (Rs, n=6) and/or were PNRs (n=6), to IFX. Serum samples were subjected to two-dimensional Gel Electrophoresis. Following evaluation of densitometrical data, protein spots exhibiting differential expression among the groups, were further characterized by MALDI-TOF-MS. Identified proteins where evaluated by immunoblot analysis while functional network association was carried out to asses significance. Proteins apolipoprotein A-I (APOA1), apolipoprotein E (APOE), complement C4-B (CO4B), plasminogen (PLMN), serotransferrin (TRFE), beta-2-glycoprotein 1 (APOH), and clusterin (CLUS) were found to be up-regulated in the PNR and Rs groups whereas their levels displayed no changes in the Rm group when compared to baseline samples. Additionally, leucine-rich alpha-2-glycoprotein (A2GL), vitamin D-binding protein (VTDB), alpha-1B-glycoprotein (A1BG) and complement C1r subcomponent (C1R) were significantly increased in the serum of the Rm group. Through the incorporation of proteomics technologies, novel serum marker-molecules demonstrating high sensitivity and specificity are introduced, hence offering an innovative approach regarding the evaluation of CD patients' response to IFX therapy. PMID:23562004

Gazouli, Maria; Anagnostopoulos, Athanasios K; Papadopoulou, Aggeliki; Vaiopoulou, Anna; Papamichael, Konstantinos; Mantzaris, Gerassimos; Theodoropoulos, George E; Anagnou, Nicholas P; Tsangaris, George Th

2013-11-01

41

Milk Serum Proteins. I. A Quantitative Biuret Test for Milk Serum Proteins1  

Microsoft Academic Search

The biuret reaction has been used for many years as a qualitative test for the presence of proteins in solution. It depends on the formation of a violet copper- protein complex in alkaline CuSQ solution. This reaction first was adapted as a quantitative test for protein by Autenrieth (1, 2), who determined the albumin and globulin in urine, ascitic fluid

B. C. Johnson; A. M. Swanson

1952-01-01

42

Unraveling the mysteries of serum albumin-more than just a serum protein.  

PubMed

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

43

Development of Gel-Filter Method for High Enrichment of Low-Molecular Weight Proteins from Serum  

PubMed Central

The human serum proteome has been extensively screened for biomarkers. However, the large dynamic range of protein concentrations in serum and the presence of highly abundant and large molecular weight proteins, make identification and detection changes in the amount of low-molecular weight proteins (LMW, molecular weight ? 30kDa) difficult. Here, we developed a gel-filter method including four layers of different concentration of tricine SDS-PAGE-based gels to block high-molecular weight proteins and enrich LMW proteins. By utilizing this method, we identified 1,576 proteins (n = 2) from 10 ?L serum. Among them, 559 (n = 2) proteins belonged to LMW proteins. Furthermore, this gel-filter method could identify 67.4% and 39.8% more LMW proteins than that in representative methods of glycine SDS-PAGE and optimized-DS, respectively. By utilizing SILAC-AQUA approach with labeled recombinant protein as internal standard, the recovery rate for GST spiked in serum during the treatment of gel-filter, optimized-DS, and ProteoMiner was 33.1 ± 0.01%, 18.7 ± 0.01% and 9.6 ± 0.03%, respectively. These results demonstrate that the gel-filter method offers a rapid, highly reproducible and efficient approach for screening biomarkers from serum through proteomic analyses. PMID:25723528

Chen, Lingsheng; Zhai, Linhui; Li, Yanchang; Li, Ning; Zhang, Chengpu; Ping, Lingyan; Chang, Lei; Wu, Junzhu; Li, Xiangping; Shi, Deshun; Xu, Ping

2015-01-01

44

The Acute-Phase Proteins Serum Amyloid A and C Reactive Protein in Transudates and Exudates  

PubMed Central

The distinction between exudates and transudates is very important in the patient management. Here we evaluate whether the acute-phase protein serum amyloid A (SAA), in comparison with C reactive protein (CRP) and total protein (TP), can be useful in this discrimination. CRP, SAA, and TP were determined in 36 exudate samples (27 pleural and 9 ascitic) and in 12 transudates (9 pleural and 3 ascitic). CRP, SAA, and TP were measured. SAA present in the exudate corresponded to 10% of the amount found in serum, that is, the exudate/serum ratio (E/S) was 0.10 ± 0.13. For comparison, the exudate/serum ratio for CRP and TP was 0.39 ± 0.37 and 0.68 ± 0.15, respectively. There was a strong positive correlation between serum and exudate SAA concentration (r = 0.764;p < 0.0001). The concentration of SAA in transudates was low and did not overlap with that found in exudates (0.02-0.21 versus 0.8–360.5 g/mL). SAA in pleural and ascitic exudates results mainly from leakage of the serum protein via the inflamed membrane. A comparison of the E/S ratio of SAA and CRP points SAA as a very good marker in discriminating between exudates and transudates. PMID:16864904

Okino, Alessandra M.; Bürger, Cristiani; Cardoso, Jefferson R.; Lavado, Edson L.; Lotufo, Paulo A.; Campa, Ana

2006-01-01

45

Relation of Serum Leptin and Adiponectin Level to Serum C-Reactive Protein: The INTERLIPID Study  

PubMed Central

Objective. Despite considerable study, the relevance of leptin and adiponectin for atherosclerosis development is still unsettled. We investigated relations of serum leptin and adiponectin to serum C-reactive protein (CRP), using the INTERLIPID dataset on Japanese emigrants living in Hawaii and Japanese in Japan. Design and Methods. Serum leptin, adiponectin, and CRP were measured by standardized methods in men and women of ages 40 to 59 years from two population samples, one Japanese-American in Hawaii (83 men, 89 women) and the other Japanese in central Japan (111 men, 104 women). Participants with CRP >10?mg/L were excluded. Results. Sex-specific multiple linear regression analyses, with log-transformed leptin and adiponectin (log-leptin, log-adipo), site (Hawaii = 1, Japan = 0), SBP, HbA1c, smoking (cigarettes/day), and physical activity index score of the Framingham Offspring Study as covariates, showed that log-leptin directly related and log-adipo inversely related to log-CRP for both sexes (Ps < 0.05 to <0.01). Addition to the model of BMI and interaction terms (BMI × log-leptin, BMI × log-adipo, SITE × log-leptin, SITE × log-adipo) resulted in disappearance of statistical significance except for direct relation of log-leptin to log-CRP in men (P = 0.006). Conclusions. Leptin directly related to CRP independent of BMI and other confounding factors in men but not in women. PMID:24371525

Nakamura, Yasuyuki; Ueshima, Hirotsugu; Miura, Katsuyuki; Kita, Yoshikuni; Okamura, Tomonori; Okayama, Akira; Choudhury, Sohel R.; Rodriguez, Beatriz; Masaki, Kamal H.; Stamler, Jeremiah

2013-01-01

46

Serum proteins in magnesium-deficient rat Y. RAYSSIGUIER P. LARVOR Y. AUGUSTI J. DURLACH  

E-print Network

Serum proteins in magnesium-deficient rat Y. RAYSSIGUIER P. LARVOR Y. AUGUSTI J. DURLACH (1 Cochin, Paris Summary. Hypoproteinemia appears early in the magnesium-deficient rat with a drop in serum. In the magnesium-deficient rat, we have shown serum protein modifications related to immunologic and allergic

Paris-Sud XI, Université de

47

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%

48

Immunoblot analysis of proteins associated with HEMA-MMA microcapsules: Human serum proteins in vitro and rat proteins following implantation  

Microsoft Academic Search

Human serum proteins and their fragments, associated with hydroxyethyl methacrylate-methyl methacrylate (HEMA-MMA) copolymer microcapsules, were characterized using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblot analysis. Capsules were incubated with serum for 1 week in vitro and then dissolved in ethanol to also precipitate the adsorbed protein. The precipitate was dissolved in 2% (w\\/v) SDS (the ‘capsule eluate’) to be

J. E. Babensee; R. M. Cornelius; J. L. Brash; M. V. Sefton

1998-01-01

49

Identification of Five Serum Protein Markers for Detection of Ovarian Cancer by Antibody Arrays  

PubMed Central

Background Protein and antibody arrays have emerged as a promising technology to study protein expression and protein function in a high-throughput manner. These arrays also represent a new opportunity to profile protein expression levels in cancer patients’ samples and to identify useful biosignatures for clinical diagnosis, disease classification, prediction, drug development and patient care. We applied antibody arrays to discover a panel of proteins which may serve as biomarkers to distinguish between patients with ovarian cancer and normal controls. Methodology/Principal Findings Using a case-control study design of 34 ovarian cancer patients and 53 age-matched healthy controls, we profiled the expression levels of 174 proteins using antibody array technology and determined the CA125 level using ELISA. The expression levels of those proteins were analyzed using 3 discriminant methods, including artificial neural network, classification tree and split-point score analysis. A panel of 5 serum protein markers (MSP-alpha, TIMP-4, PDGF-R alpha, and OPG and CA125) was identified, which could effectively detect ovarian cancer with high specificity (95%) and high sensitivity (100%), with AUC =0.98, while CA125 alone had an AUC of 0.87. Conclusions/Significance Our pilot study has shown the promising set of 5 serum markers for ovarian cancer detection. PMID:24116163

Jiang, Weidong; Huang, Ruochun; Duan, Chaohui; Fu, Liwu; Xi, Yun; Yang, Yuebo; Yang, Wei-Min; Yang, Dongzi; Yang, Dong-Hua; Huang, Ruo-Pan

2013-01-01

50

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

51

Hyperphosphatemia is a combined function of high serum PTH and high dietary protein intake in dialysis patients  

PubMed Central

Elevated serum phosphorus is associated with higher death risk in hemodialysis patients. Previous studies have suggested that both higher serum parathyroid hormone (PTH) level and higher dietary protein intake may contribute to higher serum phosphorus levels. However, it is not well known how these two factors simultaneously contribute to the combined risk of hyperphosphatemia in real patient-care scenarios. We hypothesized that the likelihood of hyperphosphatemia increases across higher serum PTH and higher normalized protein catabolic rate (nPCR) levels, a surrogate of protein intake. Over an 8-year period (July 2001–June 2009), we identified 69,355 maintenance hemodialysis patients with PTH, nPCR, and phosphorus data in a large dialysis provider. Logistic regression models were examined to assess the association between likelihood of hyperphosphatemia (serum phosphorus >5.5?mg/dl) and serum PTH and nPCR increments. Patients were 61±15 years old and included 46% women, 33% blacks, and 57% diabetics. Both higher serum PTH level and higher protein intake were associated with higher risk of hyperphosphatemia in dialysis patients. Compared with patients with PTH level 150–<300?pg/ml and nPCR level 1.0–<1.2?g/kg/day, patients with iPTH>600?pg/ml and nPCR>1.2?g/kg/day had a threefold higher risk of hyperphosphatemia (OR: 3.17, 95% CI: 2.69–3.75). Hyperphosphatemia is associated with both higher dietary protein intake and higher serum PTH level in maintenance hemodialysis patients. Worsening or resistant hyperphosphatemia may be an under-appreciated consequence of secondary hyperparathyroidism independent of dietary phosphorus load. Management of hyperphosphatemia should include diligent correction of hyper-parathyroidism while maintaining adequate intake of high protein foods with low phosphorus content. PMID:25019031

Streja, Elani; Lau, Wei Ling; Goldstein, Leanne; Sim, John J; Molnar, Miklos Z; Nissenson, Allen R; Kovesdy, Csaba P; Kalantar-Zadeh, Kamyar

2013-01-01

52

Single Nanoparticle Detection for Multiplexed Protein Diagnostics with Attomolar Sensitivity in Serum and Unprocessed Whole Blood  

E-print Network

of -lactoglobulin, a cow's milk whey protein spiked in serum (>10 orders of magnitude) and whole blood (>5 ordersSingle Nanoparticle Detection for Multiplexed Protein Diagnostics with Attomolar Sensitivity single gold nanoparticles to identify protein biomarkers in unprocessed serum and blood samples. IRIS

53

The interaction of C-reactive protein and serum amyloid P component with nuclear antigens  

Microsoft Academic Search

The pentraxins are a family of proteins characterized by cyclic pentameric structure, calcium-dependent ligand binding and sequence homology. The two main representatives of this family are the serum proteins, C-reactive protein (CRP) and serum amyloid P component (SAP). In man CRP is an acute phase reactant which increases up to 1000 fold during the acute phase response whereas SAP is

Terry W. Clos

1996-01-01

54

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

55

Small Integrin Binding Proteins as Serum Markers for Prostate Cancer Detection  

PubMed Central

Purpose The SIBLING (Small Integrin-Binding LIgand N-linked Glycoprotein) gene family includes bone sialoprotein (BSP), dentin matrix protein-1 (DMP1), dentin sialophosphoprotein (DSPP), matrix extracellular phosphoglycoprotein (MEPE), and osteopontin (OPN). Previous studies have separately reported elevated expression of BSP, OPN or DSPP in prostate tumor paraffin sections. We hypothesized that SIBLINGs may be informative serum markers for subjects with prostate cancer. Methods Expression levels of SIBLINGs in biopsies of normal tissue and tumors from prostate were determined by cDNA array and by immunohistochemical staining with monoclonal antibodies. Competitive ELISAs for measuring total BSP, DSPP, MEPE and OPN were applied to a test group of 102 subjects with prostate cancer and 110 normal subjects and a validation group of 90 subjects. Results BSP, DMP1, DSPP and OPN exhibited elevated mRNA expression and protein levels in biopsies. BSP, DSPP and OPN were elevated in serum from prostate cancer subjects, with serum DSPP exhibiting the greatest difference, yielding an area under the receiver operator characteristic curve value of 0.98. Serum BSP and OPN levels were significantly elevated only in late stages, while DSPP was significantly elevated at all stages. Optimal serum value cut-off points derived for BSP, OPN and DSPP were applied as a validation test to a new group of 90 subjects and DSPP yielded a sensitivity of 90% and a specificity of 100%. Conclusion Of the SIBLING gene family members, DSPP appears to be a strong candidate for use in serum assays for prostate cancer detection. PMID:19671866

Jain, Alka; McKnight, Dianalee A.; Fisher, Larry W.; Humphreys, Elizabeth B.; Mangold, Leslie A.; Partin, Alan W.; Fedarko, Neal S.

2009-01-01

56

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

57

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

58

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

59

A Pilot Study to Evaluate the Effect of Soy Isolate Protein on the Serum Lipid Profile and Other Potential Cardiovascular Risk Markers in Moderately Hypercholesterolemic Chinese Adults  

Microsoft Academic Search

This article examines the effect of soy isolate protein on the serum lipids and other potential cardiovascular risk markers in 90 moderately hypercholesterolemic Chinese adults (64 women and 26 men, aged 25 to 70 years). Fasting blood samples were taken before and after consuming 24 g of protein supplied by soy isolate protein supplement (including 18 g soy protein and

Lili Ma; Kerry Grann; Mei Li; Zhuoqin Jiang

2011-01-01

60

Relation between raised concentrations of fucose, sialic acid, and acute phase proteins in serum from patients with cancer: choosing suitable serum glycoprotein markers.  

PubMed Central

Serum concentrations of fucose, sialic acid, and eight acute phase proteins were measured in single specimens from patients with cancer in order to determine whether the raised concentrations of protein bound sugars commonly found in cancer correlate with increased concentrations of the acute phase proteins. Strong positive correlations were found only with alpha 1-acid glycoprotein, alpha 1-antitrypsin, and haptoglobins. Changes in protein bound sugars and acute phase proteins were also examined in relation to patients' disease states. Serum fucose was raised more often in patients with advanced disease than in those in whom the spread of the tumour was more restricted; increased sialic acid concentrations, however, were found with a similar frequency in both these groups. Combined use of fucose and sialic acid values gave a high degree of marker positivity which could be only slightly improved on by including measurement of acute phase proteins. The combined use of serum fucose and sialic acid concentrations may have value in monitoring patients with cancer: the sialic acid provides an index of the acute phase response and the fucose a measure of the tumour spread. PMID:2582003

Turner, G A; Skillen, A W; Buamah, P; Guthrie, D; Welsh, J; Harrison, J; Kowalski, A

1985-01-01

61

The serum protein fetuin-B is involved in the development of acute myocardial infarction.  

PubMed

The rupture of an atherosclerotic plaque is one of the main causes of coronary artery thrombotic occlusion, leading to myocardial infarction. However, the exact mechanism and causal risk factors for plaque rupture remain unclear. To identify a potential molecule that can influence atherosclerotic plaque rupture, we investigated protein expression in serum from patients with acute myocardial infarction (AMI) and stable angina (SA), using proteomic analysis. The expression of six proteins, including fibrinogen, fetuin-B, keratin 9, proapolipoprotein and fibrinogen, were altered in serum from patients with AMI compared with serum from those with SA. Of these, fetuin-B, proapolipoprotein, fibrinogen ?-B-chain precursors and fibrinogen expression were greater in serum from patients with AMI than from patients with SA. Increased fetuin-B expression in serum from AMI patients was also confirmed by Western blot analysis. Treatment with recombinant human fetuin-B increased the migration in monocytes and macrophages in a concentration-dependent manner. Fetuin-B also affected vascular plaque-stabilizing factors, including lipid deposition and cytokine production in macrophages, the activation of matrix metalloproteinase (MMP)-2 in monocytes, and the activation of apoptosis and MMP-2 in vascular smooth muscle cells. Moreover, in vivo administration of fetuin-B decreased the collagen accumulation and smooth muscle cell content and showed an increased number of macrophages in the vascular plaque. From these results, we suggest that fetuin-B may act as a modulator in the development of AMI. This study may provide a therapeutic advantage for patients at high risk of AMI. PMID:25671698

Jung, Seung Hyo; Won, Kyung-Jong; Lee, Kang Pa; Kim, Hyun-Joong; Seo, Eun-Hye; Lee, Hwan Myung; Park, Eun Seok; Lee, Seung Hyun; Kim, Bokyung

2015-07-01

62

Use of Composite Protein Database including Search Result Sequences for Mass Spectrometric Analysis of Cell Secretome  

PubMed Central

Mass spectrometric (MS) data of human cell secretomes are usually run through the conventional human database for identification. However, the search may result in false identifications due to contamination of the secretome with fetal bovine serum (FBS) proteins. To overcome this challenge, here we provide a composite protein database including human as well as 199 FBS protein sequences for MS data search of human cell secretomes. Searching against the human-FBS database returned more reliable results with fewer false-positive and false-negative identifications compared to using either a human only database or a human-bovine database. Furthermore, the improved results validated our strategy without complex experiments like SILAC. We expect our strategy to improve the accuracy of human secreted protein identification and to also add value for general use. PMID:25822838

Shin, Jihye; Kim, Gamin; Kabir, Mohammad Humayun; Park, Seong Jun; Lee, Seoung Taek; Lee, Cheolju

2015-01-01

63

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

64

Serum Profiles of the Regulatory Complement Proteins during the Progression of Renal Damage in Human Glomerulonephritis  

Microsoft Academic Search

Persistent activation of the complement system after escape from control by the regulatory proteins results in an excessive release of split products with inflammatory properties. Serum levels of the regulatory proteins of the complement system (Cl-INH, H, I and AT III) were assayed by the radial immunodiffusion technique in 521 serum samples from 124 patients with idiopathic chronic glomerulonephritis divided

F. P. Schena; G. Pertosa; P. Stanziale; C. Germinario; M. Balletta; V. E. Andreucci

1986-01-01

65

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

66

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

67

Study of serum C-reactive protein concentration in cardiac failure  

Microsoft Academic Search

The serum concentration of C-reactive protein was prospectively assessed in 37 patients with various degrees of heart failure. The serum concentration of C-reactive protein was higher than normal in 26 (70%) patients. The concentration was directly related to the severity of heart failure and the stage of decompensation. Hepatic cell damage is the most likely stimulus to cytokine production and

M Pye; A P Rae; S M Cobbe

1990-01-01

68

Adsorption and adhesion of common serum proteins to nanotextured gallium nitride  

NASA Astrophysics Data System (ADS)

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.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. Electronic supplementary information (ESI) available: Additional figures demonstrating the adhesion force magnitude (Fig. S1) and lateral steppe surface topography (Fig. S2). See DOI: 10.1039/c4nr06353h

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

2015-01-01

69

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

2015-02-14

70

Study of serum C-reactive protein concentration in cardiac failure.  

PubMed

The serum concentration of C-reactive protein was prospectively assessed in 37 patients with various degrees of heart failure. The serum concentration of C-reactive protein was higher than normal in 26 (70%) patients. The concentration was directly related to the severity of heart failure and the stage of decompensation. Hepatic cell damage is the most likely stimulus to cytokine production and hence release of C-reactive protein in heart failure. Heart failure is an additional cause of raised serum concentration of C-reactive protein but the pathological importance of this feature is not yet known. PMID:2337494

Pye, M; Rae, A P; Cobbe, S M

1990-04-01

71

Legionella pneumophila DNA in serum samples during Legionnaires' disease in relation to C-reactive protein levels.  

PubMed

Legionella pneumophila DNA can be detected in serum from patients with Legionnaires' disease (LD). We explored this observation studying the kinetics of L. pneumophila DNA in serum samples in relation to C-reactive protein (CRP). Eleven hospitalized patients with LD were studied. Diagnosis was made by Legionella urinary antigen test in 8 patients and seroconversion in 3 patients. A macrophage infectivity potentiator (MIP) real-time PCR was performed on 31 serum samples, including 20 follow-up serum samples. Serum samples obtained on the day of admission were MIP PCR-positive in 7 (64%) and MIP PCR-negative in 4 (36%) patients. Three (75%) of the 4 patients with a MIP PCR-negative serum sample on the day of admission became positive during follow-up. Overall, L. pneumophila DNA was detected in serum samples from 10 of the 11 patients (91%). CRP levels in the 7 patients with a positive MIP PCR serum sample on day of admission (499 +/- 144 mg/l; median +/- SD) were significantly higher than those in the 4 patients with a negative MIP PCR serum sample on the day of admission (244 +/- 97 mg/l). No difference in the severity of the disease on the day of admission was found between these patients. The presence of L. pneumophila DNA in serum is a common phenomenon in hospitalized patients with LD, although in some cases it is not yet present on the day of admission. L. pneumophila DNA in serum on the day of admission correlates with high CRP levels, but not with the severity of the disease. PMID:18855027

van de Veerdonk, F L; de Jager, C P C; Schellekens, J J A; Huijsmans, C J J; Beaumont, F; Hermans, M H A; Wever, P C

2009-04-01

72

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

73

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

74

Degeneration of Retinal ON Bipolar Cells Induced by Serum Including Autoantibody against TRPM1 in Mouse Model of Paraneoplastic Retinopathy  

PubMed Central

The paraneoplastic retinopathies (PRs) are a group of eye diseases characterized by a sudden and progressive dysfunction of the retina caused by an antibody against a protein in a neoplasm. Evidence has been obtained that the transient receptor potential melastatin 1 (TRPM1) protein was one of the antigens for the autoantibody against the ON bipolar cells in PR patients. However, it has not been determined how the autoantibody causes the dysfunction of the ON bipolar cells. We hypothesized that the antibody against TRPM1 in the serum of patients with PR causes a degeneration of retinal ON bipolar cells. To test this hypothesis, we injected the serum from the PR patient, previously shown to contain anti-TRPM1 antibodies by westerblot, intravitreally into mice and examined the effects on the retina. We found that the electroretinograms (ERGs) of the mice were altered acutely after the injection, and the shape of the ERGs resembled that of the patient with PR. Immunohistochemical analysis of the eyes injected with the serum showed immunoreactivity against bipolar cells only in wild-type animals and not in TRPM1 knockout mice,consistent with the serum containing anti-TRPM1 antibodies. Histology also showed that some of the bipolar cells were apoptotic by 5 hours after the injection in wild type mice, but no bipolar cell death was found in TRPM1 knockout mice, . At 3 months, the inner nuclear layer was thinner and the amplitudes of the ERGs were still reduced. These results indicate that the serum of a patient with PR contained an antibody against TRPM1 caused an acute death of retinal ON bipolar cells of mice. PMID:24282602

Ueno, Shinji; Nishiguchi, Koji M.; Tanioka, Hidetoshi; Enomoto, Atsushi; Yamanouchi, Takashi; Kondo, Mineo; Yasuma, Testuhiro R.; Yasuda, Shunsuke; Kuno, Noriyuki; Takahashi, Masahide; Terasaki, Hiroko

2013-01-01

75

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

76

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

77

Evaluation of the role of maternal serum high-sensitivity C-reactive protein in predicting early pregnancy failure.  

PubMed

Maternal serum high-sensitivity C-reactive protein (HSCRP) was evaluated in predicting spontaneous abortion in spontaneous pregnancies presenting with threatened spontaneous abortion. Seventy-one cases of threatened spontaneous abortion (group A) and 71 asymptomatic controls (group B), matched for gestational and maternal age, body mass index and smoking status, were included. Maternal serum samples were evaluated for HCG, progesterone, pregnancy-associated plasma protein-A (PAPP-A) and HSCRP using standard bio-assays. No difference was observed in ultrasound measurements, and median progesterone maternal serum level was significantly lower (P < 0.05) in group A compared with group B. In group A, the median of all ultrasound and maternal serum parameters was significantly lower (P < 0.01) compared with group B. The median gestational sac diameter, volume and median HSCRP and PAPP-A levels were significantly increased (P < 0.05) in group A, with a normal outcome compared with group B, probably owing to the inflammatory reaction associated with intrauterine bleeding. In group A patients destined to abortion, the gestational sac development and corresponding protein synthesis fell before the fetal heart activity stopped; in spontaneous pregnancies, maternal serum HSCRP did not provide additional information for the management of threatened spontaneous abortion but warrants further research in assisted reproduction pregnancies. PMID:25596909

Jauniaux, Eric; Gulbis, Béatrice; Jamil, Amna; Jurkovic, Davor

2015-03-01

78

Serum metabolomics reveals many novel metabolic markers of heart failure, including pseudouridine and 2-oxoglutarate  

Microsoft Academic Search

There is intense interest in the identification of novel biomarkers which improve the diagnosis of heart failure. Serum samples\\u000a from 52 patients with systolic heart failure (EF < 40% plus signs and symptoms of failure) and 57 controls were analyzed by\\u000a gas chromatography – time of flight – mass spectrometry and the raw data reduced to 272 statistically robust metabolite peaks.\\u000a 38

Warwick B. Dunn; David I. Broadhurst; Sasalu M. Deepak; Mamta H. Buch; Garry McDowell; Irena Spasic; David I. Ellis; Nicholas Brooks; Douglas B. Kell; Ludwig Neysesc

2007-01-01

79

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

80

Comparison of composition, sensory, and volatile components of thirty-four percent whey protein and milk serum protein concentrates1  

Microsoft Academic Search

The objectives of this study were to identify and compare the composition, flavor, and volatile compo- nents of serum protein concentrate (SPC) and whey protein concentrate (WPC) containing about 34% protein made from the same milk to each other and to commercial 34% WPC from 6 different factories. The SPC and WPC were manufactured in triplicate with each pair of

J. Evans; J. Zulewska; M. Newbold; M. A. Drake; D. M. Barbano

2009-01-01

81

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

82

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

PubMed Central

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

2012-01-01

83

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

84

THE ELECTROPHORETIC BEHAVIOR OF SERUM PROTEINS, OF PERCHLORIC ACID MUCOPROTEINS, AND OF THE COLLOIDAL SERUM LABILITY IN CANCEROUS TISSUES EXPOSED TO IRRADIATION  

Microsoft Academic Search

In cancer cases subjected to irradiation, thc electropho- retic behavior ; of serom proteins, perchloric acid-soluble mucoproteins, and colloid lability ; were investigated. After treatment it was observed that: (a) slight, ; statistically non-significant, alterations had occurred in the serum proteins ; (hypoalbuminaemia and hypergammaglobulinaemia accompanied by alteration in the ; albumin\\/ globulin ratio); (b) the serum mucoprotein was reduced

R. Sorrentino; E. Fumagalli

1961-01-01

85

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

PubMed

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

86

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

87

Spontaneous fluctuations in the total serum protein concentration in healthy rabbits  

Microsoft Academic Search

Regular estimation of the total blood serum protein concentration in healthy rabbits for 53 days revealed spontaneous periodic changes in this parameter. Fluctuations in the total serum protein level of the healthy rabbits has a period of 21.6  1.2 days and an amplitude of 0.41  0.10 g%, with a statistical mean level of 6.45 ~= 0.03 g%. The

M. M. Avramenko; É. N. Chirkova

1972-01-01

88

A study of serum proteins from horses infected with equine infectious anemia virus  

E-print Network

, A S 'UDY OF SERUM PROTEINS PROM HORSL'S ZNFECT D WZTH EQUZNE ZNFECTZOUS ANEMZA VZRUS A Tnesis by THOMAS MURZLZ, FOLKS Submitted to the Grs. duate College of Texas Ail:M University in partial fulfillment of the requirement for the degree... of MASTER OF SCZENCE Flay 1972 Major Subject: Veterinary Microbiolo'y A STUDY OF SERUM PROTEINS FROM HORSES INFECTED NITH EQUINE ZNFECTIOUS ANEMIA VIRUS A Thesis THOMAS MURZLL FOLKS Approved as to style and content by: o. (Chairman of Committee...

Folks, Thomas Murill

1972-01-01

89

Characterization of Serum Proteins Associated with IL28B Genotype among Patients with Chronic Hepatitis C  

PubMed Central

Introduction Polymorphisms near the IL28B gene (e.g. rs12979860) encoding interferon ?3 have recently been associated with both spontaneous clearance and treatment response to pegIFN/RBV in chronic hepatitis C (CHC) patients. The molecular consequences of this genetic variation are unknown. To gain further insight into IL28B function we assessed the association of rs12979860 with expression of protein quantitative traits (pQTL analysis) generated using open-platform proteomics in serum from patients. Methods 41 patients with genotype 1 chronic hepatitis C infection from the Duke Liver Clinic were genotyped for rs12979860. Proteomic profiles were generated by LC-MS/MS analysis following immunodepletion of serum with MARS14 columns and trypsin-digestion. Next, a latent factor model was used to classify peptides into metaproteins based on co-expression and using only those peptides with protein identifications. Metaproteins were then analyzed for association with IL28B genotype using one-way analysis of variance. Results There were a total of 4,186 peptides in the data set with positive identifications. These were matched with 253 proteins of which 110 had two or more associated, identified peptides. The IL28B treatment response genotype (rs12979860_CC) was significantly associated with lower serum levels of corticosteroid binding globulin (CBG; p?=?9.2×10?6), a major transport protein for glucocorticoids and progestins. Moreover, the CBG metaprotein was associated with treatment response (p?=?0.0148), but this association was attenuated when both IL28B genotype and CBG were included in the model, suggesting that the CBG association may be independent of treatment response. Conclusions In this cohort of chronic hepatitis C patients, IL28B polymorphism was associated with serum levels of corticosteroid binding globulin, a major transporter of cortisol, however, CBG does not appear to mediate the association of IL28B with treatment response. Further investigation of this pathway is warranted to determine if it plays a role in other comorbidities of HCV-infection. PMID:21750736

Cyr, Derek D.; Lucas, Joseph E.; Thompson, J. Will; Patel, Keyur; Clark, Paul J.; Thompson, Alexander; Tillmann, Hans L.; McHutchison, John G.; Moseley, M. Arthur; McCarthy, Jeanette J.

2011-01-01

90

Antibodies reacting with Simian Virus 40 capsid protein mimotopes in serum samples from patients affected by uveal melanoma  

PubMed Central

The uveal melanoma (UM) is the most common human intraocular tumour. Simian Virus 40 (SV-40) is a small DNA tumor virus detected in some malignancies, including the cutaneous melanoma. In this study an indirect ELISA using synthetic peptides that mimic SV-40 antigens, was employed to detect antibodies against SV-40 in serum samples from UM patients. Our report indicates a significant higher prevalence of antibodies against SV-40 capsid protein antigens in serum samples from UM patients compared to controls. Our data suggest an association between UM and SV-40, indicating that patients affected by uveal melanoma tested SV-40-positive could be treated by innovative therapies. PMID:24886631

2014-01-01

91

Identification of serum N-acetylmuramoyl-l-alanine amidase as liver peptidoglycan recognition protein 2.  

PubMed

N-acetylmuramoyl-l-alanine amidase (NAMLAA) hydrolyzes bacterial peptidoglycan and is present in human serum. A peptidoglycan-recognition protein 2 (PGLYRP2) is expressed in human liver and has N-acetylmuramoyl-l-alanine amidase activity. Here, we determined the amino acid sequences of human serum NAMLAA and liver PGLYRP2 and tested the hypothesis that serum NAMLAA and PGLYRP2 are the same protein. Liver PGLYRP2 and serum NAMLAA had the same mass determined by mass spectrometry and polyacrylamide gel electrophoresis, and both proteins and recombinant PGLYRP2 reacted with polyclonal anti-NAMLAA and anti-PGLYRP2 antibodies, and with monoclonal anti-NAMLAA antibodies. Digestion of serum NAMLAA with trypsin, chymotrypsin, or trypsin plus V8 protease, or with CNBr yielded, respectively, 37, 40, and 3 overlapping peptides that matched 100% and covered 81% of the deduced amino acid sequence of mature PGLYRP2. These peptides overlapped all exon-intron junctions indicating no alternative splice forms. Digestion of liver PGLYRP2 with trypsin yielded 23 peptides that matched 100% and covered 44% of the deduced amino acid sequence of mature PGLYRP2. Serum NAMLAA had a C398-C404 disulfide, partial phosphorylation of S218, and deamidation of N253 and N301. These results indicate that serum NAMLAA and liver PGLYRP2 are the same protein encoded by the pglyrp2 gene. PMID:16054449

Zhang, Yinong; van der Fits, Leslie; Voerman, Jane S; Melief, Marie-Jose; Laman, Jon D; Wang, Mu; Wang, Haitao; Wang, Minhui; Li, Xinna; Walls, Chad D; Gupta, Dipika; Dziarski, Roman

2005-08-31

92

Comparison of functional properties of 34% and 80% whey protein and milk serum protein concentrates.  

PubMed

This study compared the functional properties of serum protein concentrate (SPC) with whey protein concentrate (WPC) made from the same milk and with commercial WPC. The experimental SPC and WPC were produced at 34% or 80% protein from the same lot of milk. Protein contents of WPC and SPC were comparable; however, fat content was much lower in SPC compared with WPC and commercial WPC. The effect of drying methods (freeze vs. spray drying) was studied for 34% WPC and SPC. Few differences due to drying method were found in turbidity and gelation; however, drying method made a large difference in foam formation for WPC but not SPC. Between pH 3 and 7, SPC was found to have lower turbidity than WPC; however, protein solubility was similar between SPC and WPC. Foaming and gelation properties of SPC were better than those of WPC. Differences in functional properties may be explained by differences in composition and extent of denaturation or aggregation. PMID:23871371

Luck, P J; Vardhanabhuti, B; Yong, Y H; Laundon, T; Barbano, D M; Foegeding, E A

2013-09-01

93

Acute phase proteins with special reference to C-reactive protein and related proteins (pentaxins) and serum amyloid A protein.  

PubMed

The acute phase response among plasma proteins is a normal response to tissue injury and is therefore a fundamental aspect of many diverse disease processes. It probably usually has a beneficial net function in limiting damage and promoting repair but in some circumstances it may have pathological consequences. Sustained high levels of acute phase proteins and especially SAA are associated with the development of amyloidosis in some individuals. Increased concentrations of CRP may, by activating the complement system, contribute to inflammation and enhance tissue damage. Failure of the normal or appropriate CRP response may also possibly have deleterious effects. SAA is a polymorphic protein which is normally present only in trace amounts but which, during the acute phase response, becomes one of the major apolipoproteins associated with high-density lipoprotein particles. The function of apoSAA is not known but it must have considerable physiological significance apart from its role as the putative precursor of amyloid A protein fibrils. CRP and SAP have been very stably conserved throughout vertebrate evolution and homologous proteins are apparently present even in vertebrates. This strongly suggests that they have important functions although these have not yet been precisely delineated. The main role of CRP may be to provide for enhanced clearance of inappropriate materials from the plasma whether these are of extrinsic origin, such as microorganisms and their products, or the autologous products of cell damage and death. The interaction between aggregated CRP and plasma low-density lipoprotein may play a significant part in the normal function of CRP and may also have a role in lipoprotein metabolism, clearance, and deposition. SAP is a normal tissue protein as well as being a plasma protein. Aggregated SAP selectively binds fibronectin and this may represent an aspect of the normal function of SAP. The deposition of SAP in amyloid is evidently not a normal function but it is not known whether this deposition is involved in the pathogenesis of amyloid or whether it is merely an epiphenomenon. In any case immunohistochemical staining for SAP is useful in the diagnosis of amyloid, in investigation of glomerulonephritis, and in studying disorders of elastic tissue. Regardless of its physiological or pathophysiological functions, the assay of serum CRP is a valuable aid to clinical management in a number of different situations and in different diseases provided results are interpreted in the light of full clinical information.(ABSTRACT TRUNCATED AT 400 WORDS) PMID:6356809

Pepys, M B; Baltz, M L

1983-01-01

94

Correlation of serum high-sensitivity C-reactive protein and interleukin-6 in patients with acute coronary syndrome.  

PubMed

Serum high-sensitivity C-reactive protein (hs-CRP) is a sensitive indicator of inflammation, which is closely related with the progress of plaque formation. Interleukin-6 (IL-6) is one of the inflammatory markers of local coronary plaque and the peripheral blood cycle, promoting the occurrence of atherosclerosis development and plaque rupture. In this study, the correlation of hs-CRP and IL-6 was investigated in patients with acute coronary syndrome (ACS). Sixty cases of ACS, including 33 cases of acute myocardial infarction (AMI) and 27 cases of unstable angina pectoris (UAP), 45 cases of stable angina pectoris (SAP), and 45 healthy people (HG) were enrolled in study. The serum hs-CRP and serum IL-6 levels were tested by the immune turbidimetric method and enzyme-linked immunosorbent assay (ELISA), respectively. The differences among groups and their correlations were evaluated. Results showed that the serum hs-CRP and IL-6 concentrations of the AMI and UAP groups were significantly higher than those of the SAP and HG groups, respectively (P<0.01), and those of the AMI group were significantly higher than those of the UAP group (P<0.05). The serum hs-CRP and IL-6 levels of the ACS group were positively correlated (r=0.836). The serum hs-CRP and IL-6 levels could be used to determine the stability of plaque, and have some relevance in the ACS process, showing great value in judgments of ACS prognosis. PMID:25036169

Wang, X H; Liu, S Q; Wang, Y L; Jin, Y

2014-01-01

95

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

96

Total serum glycosylated proteins in detection and monitoring of gestational diabetes.  

PubMed

The goal of this study was to determine whether serum glycosylated protein levels (i.e., fructosamine) can reliably screen for gestational diabetes and whether these levels are valid markers of short-term glycemic control in the third trimester of pregnancy. Ninety-seven pregnant women at 26-28 wk gestation were evaluated over 9 mo. HbA1c and serum glycosylated protein (serum fructosamine) were determined at the baseline venipuncture of the 100-g oral glucose tolerance test performed to detect gestational diabetes. Of the 97 women studied, 13 tested positive for gestational diabetes (National Diabetes Data Group criteria). There were significant differences in the fasting and 1-, 2-, and 3-h glucose values between nondiabetic and diabetic patients (P less than 0.005 at each time point). No difference was noted in the baseline serum glycosylated protein level (2.02 +/- 0.08 vs. 1.98 +/- 0.02 mM, NS) or HbA1c level (4.42 +/- 0.2 vs. 4.6 +/- 0.3%, NS) between gestational and nondiabetic patients. Diabetic patients were followed at 2-wk intervals, with serum glycosylated protein analysis, HbA1c, fasting glucose, and mean glucose determined by outpatient monitoring. Serum glycosylated protein correlated significantly to fasting blood glucose (r = 0.81, P less than 0.001) and mean outpatient glucose (r = 0.62, P less than 0.001) at the 2-wk follow-up visits. No correlation was found between HbA1c and fasting blood glucose (r = 0.11, NS) or mean outpatient glucose (r = -0.12, NS) during the follow-up period. The serum glycosylated protein level (serum fructosamine) is not a useful screening test for gestational diabetes. However, this assay shows potential as an objective marker of short-term control in evaluating the maternal glycemic state. PMID:2209322

Cefalu, W T; Prather, K L; Chester, D L; Wheeler, C J; Biswas, M; Pernoll, M L

1990-08-01

97

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

PubMed Central

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

2015-01-01

98

Human fetal endothelial cells acquire Zinc(II) from both the protein bound and nonprotein bound pools in serum  

Microsoft Academic Search

To help determine physiologically important routes by which zinc (Zn) is acquired by human fetal vascular endothelium, the\\u000a authors incubated cultured umbilical vein endothelial cells with65Zn(II)-tracer labeled human fetal whole serum, ultrafiltrate (containing low molecular mass serum zinc complexes), and dialyzed\\u000a serum (containing protein-bound zinc). Zinc from whole serum and from both serum fractions entered a rapidly labeled cellular\\u000a compartment

Christopher M. R. Bax; David L. Bloxam

1997-01-01

99

Evaluation of histidine-rich glycoprotein tissue RNA and serum protein as novel markers for breast cancer.  

PubMed

Advances in the field of breast cancer (BC) biomarkers discovery facilitate diagnosis and treatment of BC in its pre-invasive state. While the genetic tissue markers are making significant advances in understanding the molecular basis of BC, serum has long been considered a rich source for biomarkers. So, integrated genomic and proteomic strategies play a huge role in the analytical validation of BC biomarkers and represent a true milestone in the areas of diagnostics and personalized medicine. This study included 60 cases (BC), 30 patients with fibroadenoma and 30 healthy women. Histidine-rich glycoprotein RNA (HRG) tissue expression was analyzed through gene expression-based outcome for breast cancer online algorithm (GOBO) bioinformatic analysis. To confirm our informatics analysis, HRG RNA was detected in breast tissue samples by RT-PCR, and HRG serum protein was estimated by ELISA. GOBO analysis revealed increased HRG RNA expression in all subtypes of BC with relative higher expression in basal subtype and grade 2. We confirmed these findings by HRG tissue RNA with 71.7% sensitivity and 93.3% specificity. HRG serum protein was 86.7% sensitivity and 80% specificity. HRG tissue RNA and serum protein could be considered as promising novel markers for prediction of BC prognosis. PMID:24567057

Matboli, Marwa; Eissa, Sanaa; Said, Hebatallah

2014-04-01

100

CHARACTERIZATION OF DRUG INTERACTIONS WITH SERUM PROTEINS BY USING HIGH-PERFORMANCE AFFINITY CHROMATOGRAPHY  

PubMed Central

The binding of drugs with serum proteins can affect the activity, distribution, rate of excretion, and toxicity of pharmaceutical agents in the body. One tool that can be used to quickly analyze and characterize these interactions is high-performance affinity chromatography (HPAC). This review shows how HPAC can be used to study drug-protein binding and describes the various applications of this approach when examining drug interactions with serum proteins. Methods for determining binding constants, characterizing binding sites, examining drug-drug interactions, and studying drug-protein dissociation rates will be discussed. Applications that illustrate the use of HPAC with serum binding agents such as human serum albumin, ?1-acid glycoprotein, and lipoproteins will be presented. Recent developments will also be examined, such as new methods for immobilizing serum proteins in HPAC columns, the utilization of HPAC as a tool in personalized medicine, and HPAC methods for the high-throughput screening and characterization of drug-protein binding. PMID:21395530

Hage, David S.; Anguizola, Jeanethe; Barnaby, Omar; Jackson, Abby; Yoo, Michelle J.; Papastavros, Efthimia; Pfaunmiller, Erika; Sobansky, Matt; Tong, Zenghan

2011-01-01

101

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

102

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.

103

Controlled release of bovine serum albumin from hydroxyapatite microspheres for protein delivery system  

Microsoft Academic Search

Desorption behavior of a model protein (bovine serum albumin, BSA) on commercial hydroxyapatite (HAp) microspheres and its control were investigated for protein delivery system. The desorption behavior related strongly to the phosphate concentration in phosphate buffer solution: the amount of desorbed BSA increased when the phosphate concentration increased. In physiological buffer solution, which contains 10mM phosphate, the initial burst release

Yaowalak Boonsongrit; Hiroya Abe; Kazuyoshi Sato; Makio Naito; Masahiro Yoshimura; Hideki Ichikawa; Yoshinobu Fukumori

2008-01-01

104

Vibrational circular dichroism spectra of protein films: thermal denaturation of bovine serum albumin  

Microsoft Academic Search

Vibrational circular dichroism (VCD) spectroscopy has been used for the first time to investigate the thermal denaturation of proteins in H2O solutions. Films prepared from heated aqueous solutions were used for these investigations. A well-known ?-helical protein, bovine serum albumin (BSA), is used for this first study. Both VCD and infrared absorption results obtained for BSA films indicate that the

Ganesh Shanmugam; Prasad L. Polavarapu

2004-01-01

105

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

106

Effect of bone morphogenetic protein-4 on cardiac differentiation from mouse embryonic stem cells in serum-free and low-serum media  

Microsoft Academic Search

In spite of previous reports, the precise role of bone morphogenetic proteins (BMPs) on cardiomyocyte differentiation, especially in the absence or presence of minimum amount of serum in culture medium is still unclear. So, the aim of the present study was to investigate the effect of BMP-4 on mouse embryonic stem cells (ESCs)-derived cardiomyocyte differentiation in serum-free and low-serum media.

Masoumeh Fakhr Taha; Mojtaba Rezazadeh Valojerdi

2008-01-01

107

Revision of MELD to Include Serum Albumin Improves Prediction of Mortality on the Liver Transplant Waiting List  

PubMed Central

Background Allocation of donor livers for transplantation in most regions is based on the Model for End-Stage Liver Disease (MELD) or MELD-sodium (MELDNa). Our objective was to assess revisions to MELD and MELDNa that include serum albumin for predicting waiting list mortality. Methods Adults registered for liver transplantation in the United States (2002–2007) were identified from the United Network for Organ Sharing (UNOS) database. Cox regression was used to determine the association between serum albumin and 3-month mortality, and to derive revised MELD and MELDNa scores incorporating albumin (‘MELD-albumin’ and ‘5-variable MELD [5vMELD]’). Results Among 40,393 patients, 9% died and 24% underwent transplantation within 3 months of listing. For serum albumin concentrations between 1.0 and 4.0 g/dL, a linear, inverse relationship was observed between albumin and 3-month mortality (adjusted hazard ratio per 1 g/dL reduction in albumin: 1.44; 95% CI 1.35–1.54). The c-statistics for 3-month mortality of MELD-albumin and MELD were 0.913 and 0.896, respectively (P<0.001); 5vMELD was superior to MELDNa (c-statistics 0.922 vs. 0.912, P<0.001). The potential benefit of 5vMELD was greatest in patients with low MELD (<15). Among low MELD patients who died, 27% would have gained ?10 points with 5vMELD over MELD versus only 4–7% among low MELD survivors and high MELD (?15) candidates (P<0.0005). Conclusion Modification of MELD and MELDNa to include serum albumin is associated with improved prediction of waiting list mortality. If validated and shown to be associated with reduced mortality, adoption of 5vMELD as the basis for liver allograft allocation may improve outcomes on the liver transplant waiting list. PMID:23349678

Myers, Robert P.; Shaheen, Abdel Aziz M.; Faris, Peter; Aspinall, Alexander I.; Burak, Kelly W.

2013-01-01

108

The presence of antibodies to oxidative modified proteins in serum from polycystic ovary syndrome patients  

PubMed Central

Polycystic ovary syndrome (PCOS) affects 5–10% of women of reproductive age. Free radicals, as a product of oxidative stress, impair cells and tissue properties related to human fertility. These free radicals, together with the oxidized molecules, may have a cytotoxic or deleterious effects on sperm and oocytes, on early embryo development or on the endometrium. Aldehyde-modified proteins are highly immunogenic and circulating autoantibodies to new epitopes, such as malondialdehyde (MDA), may affect the reproductive system. Autoantibodies or elevated reactive oxygen species (ROS) in serum are often associated with inflammatory response. The purpose of this work is to investigate whether PCOS women show increased levels of oxidized proteins (protein–MDA) and anti-endometrial antibodies (AEA) in their sera, compared with control patients, and to determine whether AEA specificity is related to oxidized protein derivatives. Sera from 31 women [10 patients with PCOS (PCOS group) and 21 women with male factor of infertility (control group)] were chosen from patients attending for infertility. Anti-endometrial antibodies were determined by enzyme-linked immunosorbent assay (ELISA) with an endometrial cell line (RL-95). Antibodies against MDA modified human serum albumin (HSA–MDA) were also determined by ELISA. Oxidized proteins (protein–MDA) in serum were determined by a colorimetric assay. Patients with PCOS have significantly higher levels of AEA and anti-HSA–MDA, as well as oxidized proteins (protein–MDA) in serum than control patients. For the first time, we describe an autoimmune response in PCOS patients, in terms of AEA. The evidence of protein–MDA in the serum of these patients, together with the increased antibody reactivity to MDA-modified proteins (HSA–MDA) in vitro, supports the conclusion that oxidative stress may be one of the important causes for abnormal endometrial environment with poor embryo receptivity in PCOS patients. PMID:16634794

Palacio, J R; Iborra, A; Ulcova-Gallova, Z; Badia, R; Martínez, P

2006-01-01

109

Low serum pancreatitis-associated protein does not exclude complications in mild acute pancreatitis  

Microsoft Academic Search

Normal serum PAP levels on admission to the hospital in patiens with acute pancreatitis has been proposed to help select the\\u000a patients who are not going to develop complications. The aims of this study were, first, to assess the specificity of serum\\u000a pancreatitis - associated protein (PAP) serology test and second, to evalute the usefulness of the test for prediciting

Janja Polanec; Zlatko P Pavelic; Igor Krizman; Joe Osredkar

1997-01-01

110

The clinical significance of serum soluble Fas and p53 protein in breast cancer patients: comparison with serum CA 15-3.  

PubMed

Serum sFas and p53 protein have been observed in breast cancer patients, but their clinical usefulness for diagnosis and therapy monitoring has not been clarified. The aim of this study was to compare the clinical utility of serum sFas and p53 protein with that of serum CA 15-3 as the most commonly used breast cancer tumor marker. Serum samples were taken from 35 normal healthy controls and 35 breast cancer patients before surgery, after 2 weeks of surgery and after six cycles of FAC chemotherapy. Serum sFas and p53 protein levels were measured using ELISA kits. Serum CA 15-3 levels were determined using IRMA kit. Mean Serum levels of sFas and CA 15-3 were significantly elevated while p53 protein was significantly declined in breast cancer patients than controls. Serum p53 protein showed the greatest significant area under the ROC curve (84.3%) followed by sFas (80.5%), then CA 15-3 (78%). The sensitivity, specificity and cut-off value for diagnosing breast cancer patients were 84.2%, 82.6% and 2.88 U/ml for p53 protein, 83.3%, 68.2% and 497.3 pg/ml for sFas and 45.8%, 100% and 23 U/ml for CA15-3. Surgical removal of breast resulted in a significant decline in serum sFas level with no effect on serum p53 protein and CA 15-3 levels. Six cycles of chemotherapy resulted in a significant elevation in serum sFas level with no effect on serum p53 protein and CA 15-3 levels. sFas was significantly correlated with tumor grade. It could be concluded that although serum p53 protein is superior to sFas and CA15-3 for diagnosis of breast cancer patients, only sFas is useful for monitoring the response of breast cancer patients to surgery and chemotherapy if the effect of systemic inflammatory reactions is excluded. PMID:22422199

Hewala, Taha I; Abd El-Monaim, Nadia A; Anwar, Medhat; Ebied, Samia A

2012-10-01

111

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

112

Absence of serum growth hormone binding protein in patients with growth hormone receptor deficiency (Laron dwarfism)  

SciTech Connect

It has recently been recognized that human serum contains a protein that specifically binds human growth hormone (hGH). This protein has the same restricted specificity for hGH as the membrane-bound GH receptor. To determine whether the GH-binding protein is a derivative of, or otherwise related to, the GH receptor, the authors have examined the serum of three patients with Laron-type dwarfism, a condition in which GH refractoriness has been attributed to a defect in the GH receptor. The binding of /sup 125/I-labeled hGH incubated with serum has been measured after gel filtration of the serum through an Ultrogel AcA 44 minicolumn. Results are expressed as percent of specifically bound /sup 125/I-hGH and as specific binding relative to that of a reference serum after correction is made for endogenous GH. The mean +/- SEM of specific binding of sera from eight normal adults (26-46 years of age) was 21.6 +/- 0.45%, and the relative specific binding was 101.1 +/- 8.6%. Sera from 11 normal children had lower specific binding of 12.5 +/- 1.95% and relative specific binding of 56.6 +/- 9.1%. Sera from three children with Laron-type dwarfism lacked any demonstrable GH binding, whereas sera from 10 other children with other types of nonpituitary short stature had normal relative specific binding. They suggest that the serum GH-binding protein is a soluble derivative of the GH receptor. Measurement of the serum GH-binding protein may permit recognition of other abnormalities of the GH receptor.

Daughaday, W.H.; Trivedi, B.

1987-07-01

113

Long-Term Biological Variation of Serum Protein Electrophoresis M-Spike, Urine M-Spike, and Monoclonal Serum Free Light Chain Quantification: Implications for Monitoring Monoclonal Gammopathies  

PubMed Central

BACKGROUND We analyzed serial data in patients with clinically stable monoclonal gammopathy to determine the total variation of serum M-spikes [measured with serum protein electrophoresis (SPEP)], urine M-spikes [measured with urine protein electrophoresis (UPEP)], and monoclonal serum free light chain (FLC) concentrations measured with immunoassay. METHODS Patients to be studied were identified by (a) no treatment during the study interval, (b) no change in diagnosis and <5 g/L change in serum M-spike over the course of observation; (c) performance of all 3 tests (SPEP, UPEP, FLC immunoassay) in at least 3 serial samples that were obtained 9 months to 5 years apart; (d) serum M-spike ?10 g/L, urine M-spike ?200 mg/24 h, or clonal FLC ?100 mg/L. The total CV was calculated for each method. RESULTS Among the cohort of 158 patients, 90 had measurable serum M-spikes, 25 had urine M-spikes, and 52 had measurable serum FLC abnormalities. The CVs were calculated for serial SPEP M-spikes (8.1%), UPEP M-spikes (35.8%), and serum FLC concentrations (28.4%). Combining these CVs and the interassay analytical CVs, we calculated the biological CV for the serum M-spike (7.8%), urine M-spike (35.5%), and serum FLC concentration (27.8%). CONCLUSIONS The variations in urine M-spike and serum FLC measurements during patient monitoring are similar and are larger than those for serum M-spikes. In addition, in this group of stable patients, a measurable serum FLC concentration was available twice as often as a measurable urine M-spike. PMID:21980167

Katzmann, Jerry A.; Snyder, Melissa R.; Rajkumar, S. Vincent; Kyle, Robert A.; Therneau, Terry M.; Benson, Joanne T.; Dispenzieri, Angela

2013-01-01

114

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

115

The Effect of Aerobic Exercise on Serum C - Reactive Protein and Leptin Levels in Untrained Middle-Aged Women  

PubMed Central

Background: Cardiovascular disease is the most common cause of death in the world. The aim of this study was to determine the effect of aerobic exercise on serum inflammatory markers in untrained middle-aged women. Methods: Nineteen healthy female middle-aged were selected by convenience sampling method and were randomly divided into two experimental (n=11) and control (n=8) groups. The exercise protocol included aerobic exercise training lasted for 6 months and 3 sessions per week and every session lasted for 60 minutes and with intensity of 55–65 percent of maximum heart rate reserve (MHR). Blood samples were taken to measure serum leptin and C-Reactive Protein (CRP) before and after aerobic training period. General linear-Repeated measures (GL-RM) was used to comparing of within, Interactive and between means groups. The level of significance was set at P< 0.05. Results: The level of serum leptin in middle-aged women did not change significant. However, the levels of CRP during this period did not change significantly. Conclusion: Six months of aerobic exercise does not induce significant change in serum levels of CRP, while leptin levels reduced in middle-aged women. Regular physical activity probably causes decrease in serum leptin level if body mass index and body fat mass reduce simultaneously. PMID:23193504

Bijeh, N; Hosseini, SR Attarzadeh; Hejazi, K

2012-01-01

116

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

117

Low serum sodium is associated with protein energy wasting and increased interdialytic weight gain in haemodialysis patients  

PubMed Central

Background Low serum sodium (Na) has been associated with decreased body mass index and increased cardiovascular mortality in haemodialysis (HD) patients. We examined the relationship between serum Na and selected nutritional parameters of protein energy wasting that are not affected from the hydration status in a cohort of HD patients. Methods Triceps skinfold thickness (TSF), mid-arm circumference (MAC), mid-arm muscle circumference (MAMC), handgrip strength (HGS) and subjective global assessment (SGA) were assessed in maintenance HD patients using standard techniques. MAMC was calculated with the formula MAMC (cm) = MAC (cm) ?3.142 × TSF cm. Pre-dialysis serum Na values from routine monthly laboratory measurements were averaged for the last 6 months prior to the nutritional assessment. Results Altogether 172 patients with anthropometric data were included in the final analysis. Mean age was 66 ± 14, females 62 (36%) and diabetics 48 (28.9%). Patients with pre-dialysis serum Na below the mean value (136.2 mEq/L) had lower MAMC, HGS, SGA scores and albumin levels (23.50 ± 3.16 cm versus 24.58 ± 3.71 cm, P = 0.048; 21.7 ± 13.6 kg versus 28.0 ± 12.4 kg, P = 0.030; 5.1 ± 1.2 versus 5.7 ± 1.0, P = 0.012 and 31.65 ± 4.73 mg/L versus 32.25 ± 3.91 mg/L, P = 0.022, respectively) and higher interdialytic weight gains. Pre-dialysis serum Na correlated positively with MAMC, handgrip and SGA (Pearson's correlation r = 0.165, P = 0.031, r = 0.237, P = 0.022 and r = 0.195, P = 0.011, respectively). Conclusion This study demonstrates that low serum sodium is associated with protein energy wasting and increased interdialytic weight gain in HD patients.

Poulikakos, Dimitrios; Marks, Victoria; Lelos, Nicholas; Banerjee, Debasish

2014-01-01

118

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

119

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

120

Calcium-dependent and -independent binding of the pentraxin serum amyloid P component to glycosaminoglycans and amyloid proteins: enhanced binding at slightly acid pH  

Microsoft Academic Search

Serum amyloid P component (SAP), a member of the pentraxin family of proteins, binds calcium-dependently to several ligands including glycosaminoglycans (GAG's). We have investigated the influence of pH on the Ca2+-dependent binding of SAP to solid phase GAG's and amyloid fibril proteins (AA and ?2M) by ELISA. An increase in the dose-dependent binding of SAP to heparan sulfate, AA-protein and

Bente Danielsen; Inge Juul Sørensen; Mads Nybo; Ellen Holm Nielsen; Batia Kaplan; Sven-Erik Svehag

1997-01-01

121

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

PubMed Central

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. Sera demonstrating a restricted band of protein migration (other than a definite M spike) by serum protein electrophoresis (SPE), which may represent early monoclonal proteins, were also examined. When compared to IFE, percent agreement, sensitivity, and specificity for the ?-FLC and ?-FLC were 94.6, 72.9, and 99.5% and 98.5, 91.4, and 99.7%, respectively, in detecting monoclonal proteins involving free and heavy-chain-bound light chains. The majority of sera (73.7%) demonstrating a restricted band of protein migration on SPE demonstrated abnormal IFE patterns suggestive of multiple myeloma or monoclonal gammopathy of unknown significance, but gave normal ?/? FLC ratios using the turbidimetric immunoassays. In conclusion, the ? and ? FLC assays are significantly less sensitive (72.9 to 91.4%) than IFE, but specific in detecting serum monoclonal proteins. Moreover, the ?/? ratio has little value in routine screening since the majority of sera with abnormal IFE patterns had normal ?/? FLC ratios. PMID:16467338

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

2006-01-01

122

In-Depth Analysis of a Plasma or Serum Proteome Using a 4D Protein Profiling Method  

PubMed Central

Comprehensive proteomic analysis of human plasma or serum has been a major strategy used to identify biomarkers that serve as indicators of disease. However, such in-depth proteomic analyses are challenging due to the complexity and extremely large dynamic range of protein concentrations in plasma. Therefore, reduction in sample complexity through multidimensional pre-fractionation strategies is critical, particularly for the detection of low-abundance proteins that have the potential to be the most specific disease biomarkers. We describe here a 4D protein profiling method that we developed for comprehensive proteomic analyses of both plasma and serum. Our method consists of abundant protein depletion coupled with separation strategies – microscale solution isoelectrofocusing and 1D SDS-PAGE – followed by reversed-phase separation of tryptic peptides prior to LC–MS/MS. Using this profiling strategy, we routinely identify a large number of proteins over nine orders of magnitude, including a substantial number of proteins at the low ng/mL or lower levels from approximately 300 ?L of plasma sample. PMID:21468940

Tang, Hsin-Yao; Beer, Lynn A.; Speicher, David W.

2011-01-01

123

Signaling by retinol and its serum binding protein.  

PubMed

Vitamin A, retinol, circulates in blood bound to retinol-binding protein (RBP) which, in turn, associates with transthyretin (TTR) to form a retinol-RBP-TTR ternary complex. At some tissues, retinol-bound (holo-) RBP is recognized by a membrane protein termed STRA6, which transports retinol from extracellular RBP into cells and, concomitantly, activates a JAK2/STAT3/5 signaling cascade that culminates in induction of STAT target genes. STRA6-mediated retinol transport and cell signaling are critically inter-dependent, and they both require the presence of cellular retinol-binding protein 1 (CRBP1), an intracellular retinol acceptor, as well as a retinol-metabolizing enzyme such as lecithin:retinol acyltransferase (LRAT). STRA6 thus functions as a "cytokine signaling transporter" which couples vitamin A homeostasis and metabolism to cell signaling, thereby regulating gene transcription. Recent studies provided molecular level insights into the mode of action of this unique protein. PMID:25481334

Noy, Noa

2015-02-01

124

Homology Modeling and Domain Interactions in Fetal Serum Protein  

NSDL National Science Digital Library

This exercise is intended to engage students to design, model, visualize and evaluate the theoretical three dimensional image of a protein whose structure has not yet been determined. The phylogenetic analysis in Biology Workbench of paralogs and orthologs to alphafetoprotein reveals more divergent sequences within active site domains of related proteins without biological activity and greater conservation of the alphafetoprotein active domain between different species.

Steve Festin (Hamilton College; )

2003-10-12

125

Identification of transferrin as the principal neptunium-binding protein in the blood serum of rats.  

PubMed

The binding of 239Np(V) to blood serum components of rats was examined in vivo and in vitro. After gel filtration of the serum using a Sephacryl S-300 column, 98% of the applied activity appeared with protein fractions representing coeluted albumins and transferrin. A separation of the albumin- and transferrin-proteins by ion-exchange chromatography using DEAE-cellulose showed the 239Np being entirely bound to the iron-carrier protein transferrin. The high elution yields from the ion-exchange columns, greater than 90%, suggest that the binding may be quite strong. The binding capacity of transferrin for neptunium in vivo was found to decline when the iron level in blood serum was increased. Precipitation experiments showed that 84 +/- 2% of the 239Np was precipitated with 10% (w/v) trichloracetic acid, 77 +/- 3% with 90% ethanol but only 6 +/- 1% with saturated ammonium sulphate at pH 7.4. The available data indicate that as for plutonium, thorium, americium and curium, the iron transport protein, transferrin, may be the main carrier protein for neptunium in mammalian blood serum. PMID:4086199

Wirth, R; Taylor, D M; Duffield, J

1985-01-01

126

Chickpea protein hydrolysate as a substitute for serum in cell culture  

PubMed Central

The growth of mammalian cells in vitro requires the use of rich culture media that are prepared by combining serum with specific nutrient formulations. Serum, the most expensive component of culture media, provides a complex mixture of growth factors and nutrients. Protein hydrolysates that can support in vitro cell growth and eliminate or reduce the need to use serum have been obtained from different sources. Here we describe the use of two food grade proteases to produce a chickpea protein hydrolysate that has been added to cell culture medium in order to determine whether it can be used as a substitute for serum. Medium containing the hydrolysate has been tested using two human cells lines: the monocytic THP-1 cell line which grows in suspension, and the epithelial Caco-2 cell line which grows as a monolayer. The chickpea protein hydrolysate was a good substitute for serum in the first case, but did not allow growth of Caco-2 cells. Supplementation of culture media with this inexpensive and safe hydrolysate would greatly reduce the cost of cell culture. PMID:19003183

Vioque, Javier; Pedroche, Justo; Alaiz, Manuel; Yust, María M.; Megías, Cristina; Millán, Francisco

2008-01-01

127

Chickpea protein hydrolysate as a substitute for serum in cell culture.  

PubMed

The growth of mammalian cells in vitro requires the use of rich culture media that are prepared by combining serum with specific nutrient formulations. Serum, the most expensive component of culture media, provides a complex mixture of growth factors and nutrients. Protein hydrolysates that can support in vitro cell growth and eliminate or reduce the need to use serum have been obtained from different sources. Here we describe the use of two food grade proteases to produce a chickpea protein hydrolysate that has been added to cell culture medium in order to determine whether it can be used as a substitute for serum. Medium containing the hydrolysate has been tested using two human cells lines: the monocytic THP-1 cell line which grows in suspension, and the epithelial Caco-2 cell line which grows as a monolayer. The chickpea protein hydrolysate was a good substitute for serum in the first case, but did not allow growth of Caco-2 cells. Supplementation of culture media with this inexpensive and safe hydrolysate would greatly reduce the cost of cell culture. PMID:19003183

Girón-Calle, Julio; Vioque, Javier; Pedroche, Justo; Alaiz, Manuel; Yust, María M; Megías, Cristina; Millán, Francisco

2008-07-01

128

Strategy combining separation of isotope-labeled unfolded proteins and matrix-assisted laser desorption/ionization mass spectrometry analysis enables quantification of a wide range of serum proteins.  

PubMed

A novel strategy for the quantitative profiling of serum proteome is described. It includes an ammonium sulfate depletion of the serum, an affordable stable isotope labeling chemistry for samples with a large amount of protein, separation of the unfolded proteins, and relative quantification by matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS). Labeling of unfolded proteins was performed using normal (D(0)) acrylamide and deuterated (D(3)) acrylamide. The workflow for separating the unfolded proteins includes whole gel elution and ion exchange liquid chromatography, and it combines electrophoretic separation based on the protein molecular weight followed by chromatographic separation in the presence of 8M urea based on protein charge. This was followed by trypsinolysis and MALDI MS analysis, leading to the quantification of a large number of serum proteins, including those with an abundance of 10(-5) less than albumin. This robust and inexpensive workflow is suitable for the quantitative profiling of protein changes in serum associated with preanalytical variables. PMID:18384735

Liao, Wei-Li; Turko, Illarion V

2008-06-01

129

Adsorption of bovine serum albumin on fused silica: Elucidation of protein–protein interactions by single-molecule fluorescence microscopy  

Microsoft Academic Search

The adsorption of bovine serum albumin (BSA) on fused silica at pH 4.7 was studied at the single molecules level by total-internal-reflection fluorescence microscopy. This pH value was the isoelectric point of BSA. At low [BSA] of 20pM, protein molecules adsorbed as monomers. At intermediate [BSA] of 500pM, protein molecules adsorbed as clusters of about five monomers on average. Both

K. M. Yeung; Z. J. Lu; N. H. Cheung

2009-01-01

130

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

131

Docosahexaenoic acid regulates serum amyloid A protein to promote lipolysis through down regulation of perilipin  

Microsoft Academic Search

Docosahexaenoic acid (DHA) increases lipolysis and decreases lipogenesis through several pathways. DHA also enhances the expression of serum amyloid A protein (SAA), a possible lipid metabolism related gene. The question of whether DHA regulates the expression of SAA to affect lipid metabolism and increase lipolysis needs to be demonstrated in human adipocytes. We designed experiments to determine the role of

Ya C. Wang; Wen H. Kuo; Ching Y. Chen; Hsin Y. Lin; Hsin T. Wu; Bing H. Liu; Chia H. Chen; Harry J. Mersmann; King J. Chang; Shih T. Ding

2010-01-01

132

Serum amino acid and myofibrillar protein profiles in Boer goat bucks following undernutrition  

Microsoft Academic Search

The aim of the work was to determine serum free amino acid and myofibrillar protein profiles in Boer goats following undernutrition in order to study the physiological consequences of undernutrition in this goat breed. Fifteen Boer goat bucks were allocated to two experimental groups: CG (control group), fed ad libitum Themeda trianda hay, supplemented with maize, molasses and urea; and

A. M. Almeida; L. M. J. Schwalbach; H. O. deWaal; J. P. C. Greyling; L. A. Cardoso

2004-01-01

133

Serum IGF-binding protein-6 and prostate specific antigen in breast cancer  

Microsoft Academic Search

Objective: Recent studies have demonstrated the presence of the IGF-binding proteins (IGFBPs) and prostate specific antigen (PSA), an IGFBP protease, in human breast tissue. We sought to investigate the differences in serum IGFs, IGFBP-1, -3 and -6, and PSA between patients with surgically proven breast cancer and patients with benign breast disease. Design and Methods: Concentrations of IGFs, IGFBP-1, -3

Karmal K Kaulsay; Kok-Onn Lee

1999-01-01

134

Synthetic serum substitute (SSS): A globulin-enriched protein supplement for human embryo culture  

Microsoft Academic Search

Objective: The purpose of the present study was to evaluate whether an IVF protein supplement prepared from human serum albumin (HSA) and human globulins would retain performance characteristics equivalent to those reported for the commercial plasma expanders, Plasmatein (Alpha Therapeutics, Los Angeles, California) and Plasmanate (Cutter Biological, Miles Inc., Elkhart, Indiana). Methods: Pronuclear-stage human embryos were randomly divided and cultured

Paul S. Weathersbee; Thomas B. Pool; Teri Ord

1995-01-01

135

Antibody production in packed bed reactors using serum-free and protein-free medium  

Microsoft Academic Search

The present work demonstrates the utility of packed bed reactors for the production of monoclonal antibody. We present data from a continuous process run for the production of over 100 grams of antibody, using serum-free medium. An additional pilot run also demonstrates the potential for continued antibody production under protein-free conditions, using a standard basal medium.

R. Bliem; R. Oakley; K. Matsuoka; R. Varecka; V. Taiariol

1990-01-01

136

High Throughput Quantitative Analysis of Serum Proteins using Glycopeptide Capture and Liquid Chromatography Mass Spectrometry  

SciTech Connect

It is expected that the composition of the serum proteome can provide valuable information about the state of the human body in health and disease, and that this information can be extracted via quantitative proteomic measurements. Suitable proteomic techniques need to be sensitive, reproducible and robust to detect potential biomarkers below the level of highly expressed proteins, to generate data sets that are comparable between experiments and laboratories, and have high throughput to support statistical studies. In this paper, we report a method for high throughput quantitative analysis of serum proteins. It consists of the selective isolation of peptides that are N-linked glycosylated in the intact protein, the analysis of these, no de-glycosylated peptides by LC-ESI-MS, and the comparative analysis of the resulting patterns. By focusing selectively on a few formerly N-linked glycopeptides per serum protein, the complexity of the analyte sample is significantly reduced and the sensitivity and throughput of serum proteome analysis are increased compared with the analysis of total tryptic peptides from unfractionated samples. We provide data that document the performance of the method and show that sera from untreated normal mice and genetically identical mice with carcinogen induced skin cancer can be unambiguously discriminated using unsupervised clustering of the resulting peptide patterns. We further identify, by tandem mass spectrometry, some of the peptides that were consistently elevated in cancer mice compared to their control littermates.

Zhang, Hui; Yi, Eugene C.; Li, Xiao-jun; Mallick, Parag; Kelly-Spratt, Karen S.; Masselon, Christophe D.; Camp, David G.; Smith, Richard D.; Kemp, Christopher; Aebersold, Ruedi

2005-02-01

137

Serum protein levels and neonatal growth rate of Nubian goat kids in Taiwan area  

Microsoft Academic Search

Levels of serum total protein and ?-globulin were determined in 43 newborn goat kids of up to 5 days of age on two goat farms in Taiwan using colorimetric assay and an agarose gel electrophoresis kit, respectively. The kids were sufficiently bottle-fed with the maternal colostrum every 6h for the first day and every 12h since 2 days. The overall

J. C Chen; C. J Chang; H. C Peh; S. Y Chen

1999-01-01

138

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

139

Iron Dextran treatment does not induce serum protein carbonyls in the newborn pig  

Technology Transfer Automated Retrieval System (TEKTRAN)

Oxidation of serum proteins can lead to carbonyl formation which alters their function and is often associated with stress-related diseases. Since it is recommended that all pigs reared in modern production facilities be given supplemental iron at birth to prevent anemia, and metals can catalyze th...

140

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

141

A lateral flow protein microarray for rapid determination of contagious bovine pleuropneumonia status in bovine serum  

Microsoft Academic Search

Novel analytical methods for a next generation of diagnostic devices combine attributes from sensitive, accurate, fast, simple and multiplexed analysis methods. Here, we describe a possible contribution to these by the application of a lateral flow microarray where a panel of recombinant protein antigens was used to differentiate bovine serum samples in the context of the lung disease contagious bovine

Jesper Gantelius; Carl Hamsten; Maja Neiman; Jochen M. Schwenk; Anja Persson; Helene Andersson-Svahn

2010-01-01

142

High Throughput Quantitative Analysis of Serum Proteins Using Glycopeptide Capture and Liquid Chromatography  

Microsoft Academic Search

It is expected that the composition of the serum proteome can provide valuable information about the state of the human body in health and disease and that this informa- tion can be extracted via quantitative proteomic measure- ments. Suitable proteomic techniques need to be sensi- tive, reproducible, and robust to detect potential biomarkers below the level of highly expressed proteins,

Mass Spectrometry; Hui Zhang; Eugene C. Yi; Xiao-jun Li; Parag Mallick; Karen S. Kelly-Spratt; Christophe D. Masselon; David G. Camp II; Richard D. Smith; Christopher J. Kemp; Ruedi Aebersold

143

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

144

Serum electrolyte and protein modification during different workload in jumper horse  

Microsoft Academic Search

Five clinically healthy Italian saddle horses were used to assess serum electrolyte and protein modification during different\\u000a workloads. Blood samples were collected from each horse at rest, immediately after each exercise and at 30 and 60 min after\\u000a the end of exercise. Our results confirm that exercise has variable effects, depending on work intensity, on some electrolytes,\\u000a total protein and haematocrit.

G. Piccione; C. Giannetto; A. Assenza; F. Fazio; G. Caola

2007-01-01

145

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

146

Annexin A1 protein regulates the expression of PMVEC cytoskeletal proteins in CBDL rat serum-induced pulmonary microvascular remodeling  

PubMed Central

Background Hepatopulmonary syndrome (HPS) is characterized by advanced liver disease, hypoxemia and intrapulmonary vascular dilatation (IPVD). The pathogenesis of HPS is not completely understood. Recent findings have established the role of proliferation and phenotype differentiation of pulmonary microvascular endothelial cells (PMVECs) in IPVD of HPS; the change in cytoskeletal proteins and their molecular mechanism play an essential role in the proliferation, phenotype modulation and differentiation of PMVECs. However, little is known about the relevance of cytoskeletal protein expression and its molecular mechanism in IPVD of HPS. In addition, ANX A1 protein has been identified as a key regulator in some important signaling pathways, which influences cytoskeletal remodeling in many diseases, such as lung cancer, liver cancer, etc. Methods PMVECs were cultured from the normal rats and then divided into three groups(Ad-ANXA1-transfected group, a non-transfected group, and an adenovirus empty vector group) and incubated by nomal rat serum or hepatopulmonary syndrome rat serum respectively. mRNA level was evaluated by real time reverse transcription polymerase chain reaction, and protein expression was detected by western blot. Cell proliferation was determined by the MTT and thymidine incorporation assay. Results In this study, we found that the serum from a common bile duct ligation(CBDL) Rat model decreased the expression levels of the ANX A1 mRNA and protein by at least two-fold in human PMVECs. We also found the expression of cytoskeletal proteins (Destrin, a1-actin, and a1-tubulin) in PMVECs significantly increased. After stimulating ANX A1 over-expression in PMVECs by adenovirus-mediated ANX A1 (Ad-ANXA1) transfection, we found the expression levels of cytoskeletal proteins were significantly suppressed in PMVECs at all time points. Additionally, we report here that serum from a CBDL Rat model increases the proliferation of PMVECs by nearly two-fold and that over-expression of Ad-ANXA1 significantly inhibits HPS-rat-serum-induced PMVEC proliferation (p <0.05). These findings suggest that the ANX A1 down-regulation of PMVEC proliferation in the presence of HPS-rat-serum may be the major cause of aberrant dysregulation of cytoskeletal proteins (Destrin, a1-actin, and a1-tubulin) and may, therefore, play a fundamental role in the proliferation and phenotype differentiation of PMVECs in the PVD of HPS. Conclusion Finally, the fact that transfection with Ad-ANXA1 results in inhibition of the aberrant dysregulation of cytoskeletal proteins and proliferation of PMVECs suggests a potential therapeutic effect on PVD of HPS. PMID:23587191

2013-01-01

147

Brief Characterization of Muskrat (Ondatra zibethicus) Immunoglobulin G (IgG) Separated from Serum on Protein A  

Microsoft Academic Search

Muskrat (Ondatra zibethicus) im- munoglobulin fraction was separated from whole serum by Protein A Sepharose chroma- tography. In serum electrophoresis, this frac- tion had a gamma motility; when electropho- resed on a polyactylamide gel with sodium do- decyl sulfate it resolved into two protein bands of approximately 52 and 25 kilodaltons, respec- tively. These bands were consistent with mo- lecular

Joanna D. Boruclnska; Antonio E. Garmendia

148

Prion protein detection in serum using micromechanical resonator arrays  

Microsoft Academic Search

Prion proteins that have transformed from their normal cellular counterparts (PrPc) into infectious form (PrPres) are responsible for causing progressive neurodegenerative diseases in numerous species, such as bovine spongiform encephalopathy (BSE) in cattle (also known as mad cow disease), scrapie in sheep, and Creutzfeldt–Jakob disease (CJD) in humans. Due to a possible link between BSE and CJD it is highly

Madhukar Varshney; Philip S. Waggoner; Richard A. Montagna; Harold G. Craighead

2009-01-01

149

Analysis of blood gases, serum fat and serum protein: a new approach to estimate survival chances of stranded Harbor seal (Phoca vitulina) pups from the German North Sea  

PubMed Central

Background Facing numerous challenges, such as illness, storms or human disturbance, some harbor seal (Phoca vitulina) pups lose contact to their dams and are found abandoned along the North Sea coast. In Schleswig-Holstein, pups with the prospect of surviving rehabilitation are admitted to the Seal Center Friedrichskoog. Despite elaborate clinical health assessments on admission, including differential hematology, in 2010, 17% of 108 admitted pups did not survive the first 20 days. The death rate during the years 2006 and 2009 varied between 9 and 19%. To broaden the spectrum of variables which could be predictive for survival, blood gas and serum analyses were performed for 99 pups using venous blood. Variables included total CO2, pH, partial CO2, HCO3–, base excess and anion gap as well as glucose, urea nitrogen, sodium, potassium and chloride. Moreover, total serum protein and fat (triglyceride) concentrations were measured for all pups on admission. Results Repeated measurements of 12 randomly selected individuals revealed a significant (p = 0.002) positive influence of time in rehabilitation on triglyceride concentrations. This trend probably shows the improvement of the pups’ nutritional status as a consequence of the shift from milk replacer formula to fish. No such positive influence was detected for total protein concentrations though. Hematologic values, including blood gases, were not predictive for survival. Conclusions For the first time blood gas values are reported in this study for a large sample size (N = 99) of seal pups (regardless of their health status). The ranges and medians calculated from the data can serve as a stepping stone towards the establishment of reference values for neonate harbor seals. However, future investigations on the development of blood gases in harbor seals with different health conditions and ages over time are necessary to allow for a better understanding of acid–base regulation in harbor seals. PMID:24490584

2014-01-01

150

In vitro protein binding studies with BMS-204352: lack of protein binding displacement interaction in human serum.  

PubMed

BMS-204352, a maxi-K channel opener, is currently under development for the treatment of stroke. Protein binding of BMS-204352 was determined in sera from several species, namely, rat, monkey, dog, and human. Data indicated that the compound was shown to be highly protein bound in serum from all species (ca. 99.6%). In order to test for the potential for drug-drug interactions and competitive displacement of BMS-204352 by diazepam, phenytoin, propranolol, and warfarin, in vitro experiments were performed using spiked human serum and ex vivo human plasma samples. Protein binding was determined using equilibrium dialysis for 4 h at maximal therapeutic concentrations for each drug alone or in appropriate combination in spiked serum samples. Ex vivo samples from a clinical BMS-204352 study (0, 1, and 24 h) were dialyzed separately after addition of diazepam, phenytoin, propranolol, or warfarin. Drug content in biological matrices was measured for radioactivity using liquid scintillation counting. Results indicated that (1) addition of diazepam, phenytoin, propranolol, or warfarin did not alter the free fraction of BMS-204352; (2) BMS-204352 did not displace diazepam, phenytoin, propranolol, or warfarin from their protein binding sites, and (3) comparison of ex vivo plasma samples after BMS-204352 dosing indicated no impact of BMS-204352 and/or its metabolites on the free fraction of diazepam, phenytoin, propranolol, or warfarin. In conclusion, the potential for a drug-drug interaction due to alterations in protein binding with BMS-204352 is unlikely. PMID:11745906

Krishna, R; Yao, M; Kaczor, D; Vachharajani, N; Srinivas, N R

2001-01-01

151

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

152

Serum paraoxonase activity and protein thiols in chronic renal failure patients  

PubMed Central

Serum paraoxonase is known to prevent low-density lipoprotein oxidation and atherogenesis. Association of paraoxonase with the oxidative status and lipid profile in chronic renal failure (CRF) patients on conservative management and those on chronic maintenance hemodialysis was analyzed in the present study. Serum paraoxonase, protein thiols, lipid hydroperoxides, lipid profile, creatinine and albumin levels were estimated by spectrophotometric methods in CRF patients on conservative management, those on hemodialysis and in healthy controls. Total cholesterol, triglycerides, low-density lipoprotein cholesterol, lipid hydroperoxides and creatinine levels were higher and high-density lipoprotein cholesterol, protein thiols, albumin levels and paraoxonase activity were lower in patients than in healthy controls. Paraoxonase activity correlated positively with protein thiols and high-density lipoprotein cholesterol and negatively with low-density lipoprotein cholesterol and lipid hydroperoxides. In conclusion, paraoxonase activity is decreased in CRF patients particularly on chronic maintenance hemodialysis and correlates well with the oxidative stress markers. PMID:20368914

Prakash, M.; Shetty, J. K.; Rao, L.; Sharma, S.; Rodrigues, A.; Prabhu, R.

2008-01-01

153

The Relationship between Bone Turnover and Body Weight, Serum Insulin-Like Growth Factor (IGF) I, and Serum IGF-Binding Protein Levels in Patients with Anorexia Nervosa  

Microsoft Academic Search

Malnutrition is one of the risk factors for bone loss in patients with anorexia nervosa (AN). To clarify the effects of nutritional status on bone metabolism, we examined the relationship between serum levels of nutritional indicators (insulin-like growth factor I (IGF-I), IGF- binding protein-2 (IGFBP-2), and IGFBP-3) and markers for bone metabolism (serum osteocalcin and urinary excretion of C-terminal telopeptide

MARI HOTTA; IZUMI FUKUDA; KANJI SATO; NAOMI HIZUKA; TAMOTSU SHIBASAKI; KAZUE TAKANO

2010-01-01

154

Analysis of protein oxidation in serum of fetal and newborn piglets and the influence of iron dextran on induction of protein carbonyls.  

Technology Transfer Automated Retrieval System (TEKTRAN)

Methods were employed to evaluate serum biomarkers associated with protein oxidative stress and damage, to determine potential sources of metabolic stress in baby pigs. Protein carbonyls in serum were converted to dinitrophenyl (DNP) derivatives with DNP-hydrazine, precipitated with TCA, extracted i...

155

Association between serum C- reactive protein levels and other important predictive markers of outcome in COPD.  

PubMed

C-reactive protein (CRP) is an acute-phase protein synthesized predominantly by the hepatocytes in response to tissue damage or inflammation. Levels of acute-phase proteins rise rapidly, during infection and after injury. We take up the study to correlate serum CRP levels with other important predictive markers of outcome in COPD. Patient with stable COPD (no exacerbation in the last two months) were taken up for the study. Parameters taken to correlate were age, grade of dyspnea, FEV1. It was found the CRP is negatively correlated with FEV1 and grade of dyspnea but not correlated with age. PMID:21425065

Shameem, Mohammad; Bhargava, Rakesh; Ahmad, Zuber; Saad, Talha; Fatima, Nazish; Malik, Abida

2011-01-01

156

Characterization of Granulations of Calcium and Apatite in Serum as Pleomorphic Mineralo-Protein Complexes and as Precursors of Putative Nanobacteria  

PubMed Central

Calcium and apatite granulations are demonstrated here to form in both human and fetal bovine serum in response to the simple addition of either calcium or phosphate, or a combination of both. These granulations are shown to represent precipitating complexes of protein and hydroxyapatite (HAP) that display marked pleomorphism, appearing as round, laminated particles, spindles, and films. These same complexes can be found in normal untreated serum, albeit at much lower amounts, and appear to result from the progressive binding of serum proteins with apatite until reaching saturation, upon which the mineralo-protein complexes precipitate. Chemically and morphologically, these complexes are virtually identical to the so-called nanobacteria (NB) implicated in numerous diseases and considered unusual for their small size, pleomorphism, and the presence of HAP. Like NB, serum granulations can seed particles upon transfer to serum-free medium, and their main protein constituents include albumin, complement components 3 and 4A, fetuin-A, and apolipoproteins A1 and B100, as well as other calcium and apatite binding proteins found in the serum. However, these serum mineralo-protein complexes are formed from the direct chemical binding of inorganic and organic phases, bypassing the need for any biological processes, including the long cultivation in cell culture conditions deemed necessary for the demonstration of NB. Thus, these serum granulations may result from physiologically inherent processes that become amplified with calcium phosphate loading or when subjected to culturing in medium. They may be viewed as simple mineralo-protein complexes formed from the deployment of calcification-inhibitory pathways used by the body to cope with excess calcium phosphate so as to prevent unwarranted calcification. Rather than representing novel pathophysiological mechanisms or exotic lifeforms, these results indicate that the entities described earlier as NB most likely originate from calcium and apatite binding factors in the serum, presumably calcification inhibitors, that upon saturation, form seeds for HAP deposition and growth. These calcium granulations are similar to those found in organisms throughout nature and may represent the products of more general calcium regulation pathways involved in the control of calcium storage, retrieval, tissue deposition, and disposal. PMID:19412552

Young, John D.; Young, Andrew; Hung, Chin-Ming; Young, Lena; Chao, Ying-Jie; Young, James; Wu, Cheng-Yeu

2009-01-01

157

Ultrasensitive protein detection in blood serum using gold nanoparticle probes by single molecule spectroscopy  

NASA Astrophysics Data System (ADS)

A one-step rapid and ultrasensitive immunoassay capable of detecting proteins in blood serum is developed using gold nanoprobes and fluorescence correlation spectroscopy (FCS). In this approach we take advantage of the inherent photoluminescence property of gold nanoparticles (GNPs) to develop a fluorophore-free assay to observe binding entities by monitoring the diffusion of bound versus unbound molecules in a limited confocal volume. 40-nm GNPs conjugated separately with rabbit anti-IgG (Fc) and goat anti-IgG (Fab) when incubated in blood serum containing IgG forms a sandwich structure constituting dimers and oligomers that can be differentiated by to detect IgG in blood serum at a limit of detection (LOD) of 5 pg/ml. The novelty of integrating GNPs with FCS to develop a sensitive blood immunoassay brings single molecule methods one step closer to the clinic.

Chen, Jiji; Wang, Chungang; Irudayaraj, Joseph

2009-07-01

158

Value of serum and induced sputum surfactant protein-D in chronic obstructive pulmonary disease  

PubMed Central

Background Surfactant Protein D (SP-D) is an important marker in chronic obstructive pulmonary disease (COPD). Serum SP-D levels increase while lung production of SP-D decreases in COPD. SP-D is a specific biomarker for monitoring COPD, assessment of exacerbation frequency and arrangement of treatment modalities. In the present study, we aimed to investigate the correlation between serum and induced sputum SP-D levels with severity and acute exacerbations of COPD. Method 20 healthy subjects, older than 40 years, with at least 10 pack/years smoking history (group 1), 20 stage I-II COPD patients (group 2) , and 20 stage III-IV COPD patients (group 3) were enrolled in the study. All subjects performed pulmonary function tests. Venous blood samples were taken to determine complete blood count, C-reactive protein(CRP) and serum SP-D levels. Induced sputum samples were obtained to determine SP-D level. COPD patients were followed up for acute exacerbations for 6 months. Results Serum SP-D levels of group 3 were the highest and induced sputum SP-D levels of group 2 were the lowest among the three groups. SP-D levels of induced sputum decreased in patients with increasing number of cigarette pack/years (p?=?0.03, r?=??0.115), whereas serum SP-D levels increased in these patients (p?=?0.0001, r?=?0.6 ). Induced sputum SP-D levels in COPD patients receiving inhaled corticosteroid treatment were significantly higher than in patients who were not receiving inhaler corticosteroid treatment (p?=?0.005). An inverse correlation between serum SP-D levels and FEV1 (%) was found and there was a positive correlation between the serum SP-D levels and exacerbations frequency in 6-month follow up period (p?=?0.049 ,r?=??0.252; p?=?0.0001, r?=?0.598 respectively). Conclusion Our study demonstrates the adverse effects of smoking on local SP-D levels since low levels of induced sputum SP-D were found in the group of current smokers, who were not receiving inhaled corticosteroid treatment. Relationship between serum SP-D and COPD exacerbations frequency suggests that serum SP-D level may be used as a lung-specific biomarker during the follow up and progression of COPD. PMID:23725346

2013-01-01

159

Effects of obstructive sleep apnea on serum brain-derived neurotrophic factor protein, cortisol, and lipid levels  

Microsoft Academic Search

Objectives  Obstructive sleep apnea (OSA) is a sleep-disordered breathing leading to vascular endothelial cells dysfunction, cognitive\\u000a impairment, and abnormal lipid metabolism. serum brain-derived neurotrophic factor (BDNF) protein, cortisol, and lipid levels\\u000a in OSA were investigated.\\u000a \\u000a \\u000a \\u000a \\u000a Materials and methods  All middle-aged subjects including healthy individuals without signs and symptoms of apnea-hypopnea and ear nose throat (ENT)\\u000a outpatients were randomly recruited and screened by

Busarakumtragul Panaree; Mekseepralard Chantana; Sukhumsirichart Wasana; Neruntarat Chairat

160

MALDI-TOF-MS serum protein profiling for developing diagnostic models and identifying serum markers for discogenic low back pain  

PubMed Central

Background The identification of the cause of chronic low back pain (CLBP) represents a great challenge to orthopedists due to the controversy over the diagnosis of discogenic low back pain (DLBP) and the existence of a number of cases of CLBP of unknown origin. This study aimed to develop diagnostic models to distinguish DLBP from other forms of CLBP and to identify serum biomarkers for DLBP. Methods Serum samples were collected from patients with DLBP, chronic lumbar disc herniation (LDH), or CLBP of unknown origin, and healthy controls (N), and randomly divided into a training set (n?=?30) and a blind test set (n?=?30). Matrix-assisted laser desorption ionization time-of-flight mass spectrometry was performed for protein profiling of these samples. After the discriminative ability of two most significantly differential peaks from each two groups was assessed using scatter plots, classification models were developed using differential peptide peaks to evaluate their diagnostic accuracy. The identity of peptides corresponding to three representative differential peaks was analyzed. Results The fewest statistically significant differential peaks were identified between DLBP and CLBP (3), followed by CLBP vs. N (5), DLBP vs. N (9), LDH vs. CLBP (20), DLBP vs. LDH (23), and LDH vs. N (43). The discriminative ability of two most significantly differential peaks was poor in classifying DLBP vs. CLBP but good in classifying DLBP vs. LDH. The accuracy of models for classification of DLBP vs. CLBP was not very high in the blind test (forecasting ability, 67.24%; sensitivity, 70%), although a higher accuracy was observed for classification of DLBP vs. LDH and LDH vs. N (forecasting abilities, ~90%; sensitivities, >90%). A further investigation of three representative differential peaks led to the identification of two peaks as peptides of complement C3, and one peak as a human fibrinogen peptide. Conclusions Our findings benefit not only the diagnosis of CLBP but also the understanding of the differences between different forms of DLBP. The ability to distinguish between different causes of CLBP and the identification of serum biomarkers may be of great value to diagnose different causes of DLBP and predict treatment efficacy. PMID:24889399

2014-01-01

161

Measuring interactions between polydimethylsiloxane and serum proteins at the air-water interface.  

PubMed

The interaction between synthetic polymers and proteins at interfaces is relevant to basic science as well as a wide range of applications in biotechnology and medicine. One particularly common and important interface is the air-water interface (AWI). Due to the special energetics and dynamics of molecules at the AWI, the interplay between synthetic polymer and protein can be very different from that in bulk solution. In this paper, we applied the Langmuir-Blodgett technique and fluorescence microscopy to investigate how the compression state of polydimethylsiloxane (PDMS) film at the AWI affects the subsequent adsorption of serum protein [e.g., human serum albumin (HSA) or immunoglobulin G (IgG)] and the interaction between PDMS and protein. Of particular note is our observation of circular PDMS domains with micrometer diameters that form at the AWI in the highly compressed state of the surface film: proteins were shown to adsorb preferentially to the surface of these circular PDMS domains, accompanied by a greater than 4-fold increase in protein found in the interfacial film. The PDMS-only film and the PDMS-IgG composite film were transferred to cover glass, and platinum-carbon replicas of the transferred films were further characterized by scanning electron microscopy and atomic force microscopy. We conclude that the structure of the PDMS film greatly affects the amount and distribution of protein at the interface. PMID:23819833

Liao, Zhengzheng; Hsieh, Wan-Ting; Baumgart, Tobias; Dmochowski, Ivan J

2013-07-30

162

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. PMID:25667676

LIU, WEI; ZHAO, JING; WEI, YUJUAN

2015-01-01

163

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

164

Cytocidal effect of gossypol on cultured murine erythroleukemia cells is prevented by serum protein.  

PubMed

The interaction of gossypol (G) with cultured murine erythroleukemia cells (MELC) was studied in vitro. G was cytocidal (inhibited growth greater than 90%) to MELC at greater than 10 microM, but not at less than 5 microM in medium supplemented with 10 and 15% fetal calf serum (FCS). Five micromolar of G was cytocidal in 2 and 5% FCS. Serum albumin (2%) also decreased the effective cytocidal dose of G. This inhibition was reversible if extracellular drug (30 microM) was removed after 1 h but not after 24 h. Uptake of [14C]G by MELC (approximately 10(6) cells/ml) was saturable with half-maximal uptake at 8 microM in the absence of FCS. This uptake was concentrative, i.e., 75-fold relative to the total [14C]G concentration. In the presence of both FCS (approximately 2%) and serum albumin (approximately 0.03%), the accumulation of [14C]G (10 microM) by MELC was decreased approximately 50%. Direct binding of G to albumin, assayed by quenching of intrinsic fluorescence, was stoichiometric with respect to micromolar albumin content suggesting an apparent affinity of approximately 10(-7) M. Visible absorption spectra for G in the presence of serum albumin exhibited batho- and hyperchromic shifts of the maxima at approximately 380 nm. These studies demonstrate that: 1) G, at pharmacologically relevant concentrations, is cytocidal for MELC; 2) serum protein, e.g., albumin, can reduce both the cytocidal effects of G and the uptake of [14C]G by MELC; and 3) these effects are probably the result of G binding to serum protein, e.g., albumin, which reduces the free effective concentration of the drug.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:6584595

Haspel, H C; Ren, Y F; Watanabe, K A; Sonenberg, M; Corin, R E

1984-04-01

165

Fetal bovine serum xenoproteins modulate human monocyte adhesion and protein release on biomaterials in vitro  

PubMed Central

Monocyte-derived macrophages are critical in the host foreign body response to biomaterials and have been studied extensively in various culture conditions in vitro such as medium supplemented with fetal bovine serum (FBS) or autologous human serum (AHS). Since monocyte maturation into macrophages is highly plastic and may vary considerably depending on the surface, isolation procedures, and in vitro culture conditions, we hypothesize that variations in protein adsorption and serum type will greatly impact monocyte behavior in a surface-dependent manner. The impact of xenoproteins on monocyte-surface interaction is not well studied methodically and the use of AHS rather than FBS for macrophage-biomaterials studies in vitro is far from universal. The commonly used reference materials: tissue culture polystyrene (TCPS), polyethylene glycol (PEG), and poly-dimethylsiloxane (PDMS) were employed in this study and we found a 3-fold higher adherent monocyte density on TCPS when AHS was used versus FBS-supplemented medium. On PEG hydrogels, an 8-10 fold higher adhesion density was observed when AHS was employed versus FBS, while on PDMS no difference in adhesion density was observed between the two sera conditions. Additionally, the presence of lipopolysaccharide abrogated the serum-dependent effect on cell adhesion on TCPS. Significant differential variations in protein release were observed between the serum conditions on these surfaces, in particular there was a 100-fold higher concentration of growth-related oncogene for the AHS condition on PDMS even though the adhesion levels were comparable between the two serum conditions. These results emphasize the combined impact of the surface type and FBS xenoproteins in mediating the observed monocyte response to biomaterials in vitro. PMID:20837169

Schmidt, David; Joyce, Evan James; Kao, Weiyuan John

2010-01-01

166

Polymorphism of rs873308 near the transmembrane protein 57 gene is associated with serum lipid levels  

PubMed Central

SNP (single-nucleotide polymorphism) of rs10903129 near the TMEM (transmembrane protein) 57 locus has been associated with TC (total cholesterol) in a previous GWAS (genome-wide association study), but the association of TMEM57 rs873308 SNP and serum lipid levels has not been previously reported. The current study was undertaken to detect the association of the TMEM57 rs873308 SNP and several environmental factors with serum lipid profiles in the Han Chinese and Mulao populations. The genotypes of the TMEM57 rs873308 SNP in 865 individuals of Han Chinese and 902 participants of Mulao nationality were determined by PCR and RFLP (restriction-fragment-length polymorphism) combined with gel electrophoresis and then confirmed by direct sequencing. The T allele frequency of TMEM57 rs873308 SNP was not different between Han and Mulao (23.18% versus 25.72%, P>0.05), but different between males and females in the two ethnic groups (P<0.05). The T allele carriers had lower serum TC, Apo (apolipoprotein) B, HDL-C (high-density lipoprotein cholesterol) levels, ApoA1/ApoB ratio in Han; and lower TAG (triacylglycerol), LDL-C (low-density lipoprotein cholesterol), ApoA1 levels and the ApoA1/ApoB ratio and higher HDL-C levels in Mulao than the T allele non-carriers. There was also different association of the TMEM57 rs873308 SNP and serum lipid profiles between males and females in the both ethnic groups. Serum lipid parameters in the two ethnic groups were also associated with several environmental factors. The association of the TMEM57 rs873308 SNP and serum lipid levels was different in the Han Chinese and Mulao populations and between males and females in the both ethnic groups. There may be a sex-specific association of the TMEM57 rs873308 SNP and serum lipid levels in our study populations. PMID:24517463

Guo, Tao; Yin, Rui-Xing; Lin, Quan-Zhen; Wu, Jian; Shen, Shao-Wen; Sun, Jia-Qi; Shi, Guang-Yuan; Wu, Jin-Zhen; Li, Hui; Wang, Yi-Ming

2014-01-01

167

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)

2001-11-13

168

Serum protein layers on parylene-C and silicon oxide: effect on cell adhesion.  

PubMed

Among the range of materials used in bioengineering, parylene-C has been used in combination with silicon oxide and in presence of the serum proteins, in cell patterning. However, the structural properties of adsorbed serum proteins on these substrates still remain elusive. In this study, we use an optical biosensing technique to decipher the properties of fibronectin (Fn) and serum albumin adsorbed on parylene-C and silicon oxide substrates. Our results show the formation of layers with distinct structural and adhesive properties. Thin, dense layers are formed on parylene-C, whereas thicker, more diffuse layers are formed on silicon oxide. These results suggest that Fn acquires a compact structure on parylene-C and a more extended structure on silicon oxide. Nonetheless, parylene-C and silicon oxide substrates coated with Fn host cell populations that exhibit focal adhesion complexes and good cell attachment. Albumin adopts a deformed structure on parylene-C and a globular structure on silicon oxide, and does not support significant cell attachment on either surface. Interestingly, the co-incubation of Fn and albumin at the ratio found in serum, results in the preferential adsorption of albumin on parylene-C and Fn on silicon oxide. This finding is supported by the exclusive formation of focal adhesion complexes in differentiated mouse embryonic stem cells (CGR8), cultured on Fn/albumin coated silicon oxide, but not on parylene-C. The detailed information provided in this study on the distinct properties of layers of serum proteins on substrates such as parylene-C and silicon oxide is highly significant in developing methods for cell patterning. PMID:25555155

Delivopoulos, Evangelos; Ouberai, Myriam M; Coffey, Paul D; Swann, Marcus J; Shakesheff, Kevin M; Welland, Mark E

2015-02-01

169

Serum protein layers on parylene-C and silicon oxide: Effect on cell adhesion  

PubMed Central

Among the range of materials used in bioengineering, parylene-C has been used in combination with silicon oxide and in presence of the serum proteins, in cell patterning. However, the structural properties of adsorbed serum proteins on these substrates still remain elusive. In this study, we use an optical biosensing technique to decipher the properties of fibronectin (Fn) and serum albumin adsorbed on parylene-C and silicon oxide substrates. Our results show the formation of layers with distinct structural and adhesive properties. Thin, dense layers are formed on parylene-C, whereas thicker, more diffuse layers are formed on silicon oxide. These results suggest that Fn acquires a compact structure on parylene-C and a more extended structure on silicon oxide. Nonetheless, parylene-C and silicon oxide substrates coated with Fn host cell populations that exhibit focal adhesion complexes and good cell attachment. Albumin adopts a deformed structure on parylene-C and a globular structure on silicon oxide, and does not support significant cell attachment on either surface. Interestingly, the co-incubation of Fn and albumin at the ratio found in serum, results in the preferential adsorption of albumin on parylene-C and Fn on silicon oxide. This finding is supported by the exclusive formation of focal adhesion complexes in differentiated mouse embryonic stem cells (CGR8), cultured on Fn/albumin coated silicon oxide, but not on parylene-C. The detailed information provided in this study on the distinct properties of layers of serum proteins on substrates such as parylene-C and silicon oxide is highly significant in developing methods for cell patterning. PMID:25555155

Delivopoulos, Evangelos; Ouberai, Myriam M.; Coffey, Paul D.; Swann, Marcus J.; Shakesheff, Kevin M.; Welland, Mark E.

2015-01-01

170

A novel protein isolated from the serum of rabbitfish (Siganus oramin) is lethal to Cryptocaryon irritans.  

PubMed

The susceptibility of eight marine fish species cultured in South China were tested for infection by the parasitic ciliate, Cryptocaryon irritans, via a challenge examination and an immobilization assay. All species of fish (representing six different families) that we investigated were infected by C. irritans except the rabbitfish (Siganus oramin), which displayed resistance to C. irritans infection. The infection intensity of rabbitfish (0.92+/-0.97, p<0.05) was significantly lower while the immobilization titres of rabbitfish serum were significantly higher (44.51+/-22.98, p<0.05) than the other seven species of fish. Additionally, the serum of the rabbitfish presented a strong killing effect to C. irritans in vitro. Light microscopy, scanning electron microscopy and fluorescence microscopy confirmed that rabbitfish serum could induce the theront cilia fall off, rupture of the cell membrane because of the swell and rupture of the macronucleus. Rabbitfish serum could also induce the rupture of the trophont membrane and content efflux. Herein a novel antiparasitic protein (APP) was isolated and purified from the serum of rabbitfish (S. oramin) by using a series of salting-out, cation exchange chromatography and two step of reversed phase high performance liquid chromatography (RP-HPLC). Analysis of sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) showed that APP was a homogenous polymeric protein with an N-terminal amino acid sequence of SSVEKNLAACLRDND. Its monomeric molecular mass, determined by matrix-assisted laser desorption ionization tandem time-of-flight mass spectrometer (MALDI-TOF-TOF-MS), was found to be 61,739.87 Da. Results of homology analyses indicated that this protein was a newly discovered functional protein in the rabbitfish serum. Laser confocal fluorescence microscopy conformed that the action site of the APP was mainly on the cell membrane and nucleus of theront, which agreed with the results of light microscopy, fluorescence microscopy and scanning electron microscopy. These findings suggest that this protein may contribute considerably to the innate host defence mechanism to combat microbes of the rabbitfish. PMID:20117218

Wang, Fang-Hua; Xie, Ming-Quan; Li, An-Xing

2010-07-01

171

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

172

Effects of egg white protein supplementation on muscle strength and serum free amino acid concentrations.  

PubMed

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

2012-10-01

173

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

174

Serum insulin-like growth factors and their binding proteins in children with end-stage renal disease  

Microsoft Academic Search

Serum levels of insulin-like growth factor-I (IGF-I), IGF-II, and IGF binding protein-1 (IGFBP-1), IGFBP-2, and IGFBP-3 were measured in 54 children with end-stage renal disease (ESRD). The results were compared with their respective age-dependent normal ranges. IGFs and IGFBPs were quantified by specific radioimmunoassay. Serum IGF-I in children with ESRD tended to cluster in the low-normal range. Mean age-related serum

Burkhard Tönshoff; Werner F. Blum; Otto Mehls

1996-01-01

175

Human growth hormone expressed in tobacco cells as an arabinogalactan-protein fusion glycoprotein has a prolonged serum life  

Microsoft Academic Search

Therapeutic proteins with molecular weights lower than 40 kDa often have short serum half-lives due to their susceptibility\\u000a to serum proteases and rapid renal clearance. Chemical derivatization, such as PEGylation, or expression as serum albumin\\u000a fusions increases molecular mass and overcome these problems but at the expense of decreased bioactivity. Here we applied\\u000a a new method that yields biologically potent recombinant

Jianfeng Xu; Shigeru Okada; Li Tan; Kenneth J. Goodrum; John J. Kopchick; Marcia J. Kieliszewski

2010-01-01

176

Inflammatory Serum Proteins Are Severely Altered in Metastatic Gastric Adenocarcinoma Patients from the Chinese Population  

PubMed Central

Background Inflammation is one of the major hallmarks of cancer. This study was designed to profile a panel of inflammatory mediators in gastric adenocarcinoma (GA) and to identify their potential differences separately in metastatic and non-metastatic patient subgroups. Methods Serum samples from 216 GA patients and 333 healthy controls from China were analyzed for six proteins using the Luminex multiplex assay. Results The serum levels for all the six proteins were significantly elevated in metastatic GA compared to non-metastatic GA. Two acute phase proteins (SAA and CRP) and a CXC chemokine (GRO) were significantly elevated in metastatic GA (p <0.01) but smaller changes were observed in non-metastatic GA compared to healthy controls. OPN is moderately increased in non-metastatic GA (2.05-fold) and more severely elevated in metastatic GA (3.34-fold). Surprisingly, soluble VCAM1 and AGP were significantly lower in both non-metastatic and metastatic GA patients compared to controls. Several individual proteins were shown to possess moderate diagnostic value for non-metastatic GA (AUC = 0.786, 0.833, 0.823 for OPN, sVCAM1 and AGP, respectively) and metastatic GA (AUC = 0.931, 0.720, 0.834 and 0.737 for OPN, sVCAM1, SAA and CRP, respectively). However, protein combinations further improve the diagnostic potential for both non-metastatic GA (best AUC = 0.946) and metastatic GA (best AUC = 0.963). The protein combination with best AUC value for both comparisons is OPN+sVCAM1+AGP+SAA. Conclusions These results suggest that several serum proteins are directly related to the severity of gastric cancer. Overall, stronger associations are observed with metastatic than non-metastatic GA as the protein changes are greater with the metastatic status. A combination of these serum proteins may serve as non-invasive markers to assess the severity status and stage of gastric cancer. PMID:25884401

Sharma, Ashok; He, Mingfang; Xue, Jing; Wu, Jianzhong; Dun, Boying; Li, Gang; Wang, Xiaoxiao; Ji, Minghua; She, Jin-Xiong; Tang, Jinhai

2015-01-01

177

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

178

Serum monocyte chemoattractant protein-1 levels in rat models of intimal hyperplasia  

Microsoft Academic Search

Recently we reported that there is a direct correlation between monocytes\\/macrophages (Mo\\/Mø) infiltration and the development of intimal hyperplasia (IH) in rat interposition vein graft. Monocyte chemoattractant protein-1 (MCP-1) is the most potent chemoattractant and activating chemokine for Mo\\/Mø. We evaluated rat serum MCP-1 levels as the indicator of inflammatory response, before and after operation. In twenty five male Lewis

Mitsuhiro Yamamura; Takashi Miyamoto; Hideki Yao

2002-01-01

179

Acute-Phase Proteins during Hemodialysis: Correlations with Serum Interleukin1? Levels and Different Dialysis Membranes  

Microsoft Academic Search

The effects of hemodialysis (HD) on the levels of serum amyloid A (SAA), C-reactive protein (CRP) and interleukin-1? (IL-1?) were studied in 8 patients. Bicarbonate dialysate was used exclusively, and three different membranes, Cuprophan® (CU), cellulose acetate (CA), and polymethylmetachrylate (PMMA) were compared. The SAA levels increased significantly with each membrane. With CU, they rose from 4.0 ± 2.0 (mg\\/l,

Eero Honkanen; Carola Grönhagen-Riska; Anna-Maija Teppo; C. P. J. Maury; Seppo Meri

1991-01-01

180

Evaluation of serum protein separation by capillary electrophoresis: prospective analysis of 1000 specimens  

Microsoft Academic Search

To critically assess the method of capillary electrophoresis (CE) we examined 1000 prospective serum samples submitted for protein electrophoresis by both high-resolution agarose gel electrophoresis (HRAGE) and CE. CE was performed using a 72 cm (50 cm to detector) × 50 ?m I.D. fused-silica capillary with detection of absorbance at 200 nm. The 1000 samples examined contained 362 monoclonal paraproteins

Margaret A. Jenkins; Elena Kulinskaya; Helen D. Martin; Michael D. Guerin

1995-01-01

181

Serum retinol binding protein 4 contributes to insulin resistance in obesity and type 2 diabetes  

Microsoft Academic Search

In obesity and type 2 diabetes, expression of the GLUT4 glucose transporter is decreased selectively in adipocytes. Adipose-specific Glut4 (also known as Slc2a4) knockout (adipose-Glut4-\\/-) mice show insulin resistance secondarily in muscle and liver. Here we show, using DNA arrays, that expression of retinol binding protein-4 (RBP4) is elevated in adipose tissue of adipose-Glut4-\\/- mice. We show that serum RBP4

Qin Yang; Timothy E. Graham; Nimesh Mody; Frederic Preitner; Odile D. Peroni; Janice M. Zabolotny; Ko Kotani; Loredana Quadro; Barbara B. Kahn

2005-01-01

182

Effects of dietary fats and proteins on rat testicular steroidogenic enzymes and serum testosterone levels.  

PubMed

It is known that certain dietary fats can modulate rat testosterone metabolism. In the current study we have investigated testicular steroidogenic enzyme activities and serum testosterone levels in rats fed diets containing either different protein sources (casein, fishmeal, whey) or different lipid sources (soybean oil, docosahexaenoic acid (DHA), seal oil, fish oil, lard). The diets examined reflect different marine oils and proteins which are significant components of Northern Canadian diets. Male rats (42-45 days old, 6 per group), were assigned to specific diets for 42 days. On the 43rd day of the study, rats were sacrificed and blood plasma and testes frozen (-80 degrees C) until analysis. Microsomal steroidogenic enzyme activities (3beta-HSD, 17-OHase, C-17,20-lyase, 17beta-HSD) were measured radiometrically. There were no differences in enzyme activities between the three dietary protein sources. In contrast, compared with the standard casein diet, all lipid sources caused reductions in C-17,20-lyase activity (>50%); seal oil and fish oil reduced 17-OHase activity (approximately 30%) and soybean oil, DHA fish oil and lard reduced 17beta-HSD activity (approximately 30%). No effect on 3beta-HSD activity was evident. Serum testosterone levels were determined using ELISA kits and were not affected by any diet with the exception of the soybean oil diet which was significantly elevated compared with the casein protein diet. Body and testis weights were not affected by diet. In conclusion, these data demonstrate that some dietary lipid sources caused reductions in testicular 17-OHase and C-17,20-lyase activities but not to the extent that serum T levels were affected, while soybean oil caused elevated serum testosterone in the absence of elevated steroidogenic enzyme activities. PMID:17936465

McVey, Mark J; Cooke, Gerard M; Curran, Ivan H A; Chan, Hing Man; Kubow, Stan; Lok, Eric; Mehta, Rekha

2008-01-01

183

Evaluation of IGF-I levels and serum protein profiles of diabetic cats and dogs  

PubMed Central

In this study, we measured the insulin-like growth factor (IGF)-I levels and evaluated the serum protein profiles of diabetic, insulin-treated, and healthy cats and dogs. The total IGF-I concentrations were 33.74 ± 3.4 ng/mL for normal, 25.8 ± 4.5 ng/mL for diabetic, and 180.4 ± 31.4 ng/mL for insulin-treated cats. IGF-I concentrations were 46.4 ± 6.6 ng/mL for normal, 25.1 ± 4.1 ng/mL for diabetic, and 303.0 ± 61.3 ng/mL for insulin-treated dogs. Total serum protein profiles were analyzed by SDS-PAGE. Fourteen bands ranging from 25 to 240 kDa in size were observed for cats, and 17 bands ranging from 25 to 289 kDa were observed for dogs. The densities of the bands differed among control, diabetic, and insulin-treated animals. In conclusion, we found that serum protein profiles and IGF-I concentrations were altered in both diabetic and insulin-treated animals. When judiciously interpreted in the light of other clinical and laboratory data, the techniques used in our study provide a valuable modality for measuring the severity of diabetes mellitus in dogs and cats. PMID:22122898

Yarim, Gul Fatma

2011-01-01

184

Evaluation of IGF-I levels and serum protein profiles of diabetic cats and dogs.  

PubMed

In this study, we measured the insulin-like growth factor (IGF)-I levels and evaluated the serum protein profiles of diabetic, insulin-treated, and healthy cats and dogs. The total IGF-I concentrations were 33.74 ± 3.4 ng/mL for normal, 25.8 ± 4.5 ng/mL for diabetic, and 180.4 ± 31.4 ng/mL for insulin-treated cats. IGF-I concentrations were 46.4 ± 6.6 ng/mL for normal, 25.1 ± 4.1 ng/mL for diabetic, and 303.0 ± 61.3 ng/mL for insulin-treated dogs. Total serum protein profiles were analyzed by SDS-PAGE. Fourteen bands ranging from 25 to 240 kDa in size were observed for cats, and 17 bands ranging from 25 to 289 kDa were observed for dogs. The densities of the bands differed among control, diabetic, and insulin-treated animals. In conclusion, we found that serum protein profiles and IGF-I concentrations were altered in both diabetic and insulin-treated animals. When judiciously interpreted in the light of other clinical and laboratory data, the techniques used in our study provide a valuable modality for measuring the severity of diabetes mellitus in dogs and cats. PMID:22122898

Ciftci, Gulay; Yarim, Gul Fatma

2011-12-01

185

Temporal changes (1997-2012) of perfluoroalkyl acids and selected precursors (including isomers) in Swedish human serum.  

PubMed

Concentrations (including isomer patterns) and temporal changes (1997-2012) of perfluoroalkyl acids (PFAAs) and selected perfluorooctane sulfonate (PFOS) and perfluoroalkyl carboxylic acid (PFCA) precursors were determined in serum samples from Swedish women. Perfluorooctane sulfonamide (FOSA) and perfluorooctane sulfonamidoacetic acid (FOSAA), as well as its N-methyl and N-ethyl derivatives (MeFOSAA and EtFOSAA) were consistently detected. Highest PFOS precursor concentrations were found for EtFOSAA (before year 2000) or MeFOSAA and FOSAA (after 2000). Disappearance half-lives for all PFOS precursors were shorter compared to PFOS. 4:2/6:2 and 6:2/6:2 polyfluoroalkyl phosphate diesters (diPAPs) were detected in <60% of the samples, whereas 6:2/8:2 and 8:2/8:2 diPAPs were detected in >60% of the samples, but showed no significant change in concentrations over time. Linear and sum-branched isomers were quantified separately for three PFAAs and three precursors. Significant changes between 1997 and 2012 in the % linear isomer were observed for PFOA and FOSA (increase) and PFOS (decrease). PMID:25660070

Gebbink, Wouter A; Glynn, Anders; Berger, Urs

2015-04-01

186

Levels of serum superoxide dismutase and high sensitive C-reactive protein in type 2 diabetic patients with lower extremity vascular disease are enhanced by interventional treatment  

PubMed Central

Objectives: This study is to determine the levels of serum superoxide dismutase (SOD) and high sensitive C-reactive protein (hs-CRP) in type 2 diabetic patients with lower extremity vascular disease before and after interventional treatment. Methods: A total of 65 patients were enrolled in this study, including 35 mails and 30 females. Another 65 healthy individuals were used as control, including 41 males and 24 females. Lesions and degrees of stenosis were determined by computed tomography angiography. Contralateral iliac artery and proximal femoral artery occlusion were treated by retrograde femoral artery puncture. The levels of serum SOD and hs-CRP were determined by enzyme-linked immunosorbent assay. Correlation was analyzed by Pearson’s test. Progression-free survival curve was analyzed by Kaplan-Meier method. Results: The levels of serum SOD at 20 min, 24 hr, 7 d, and 14 d after surgery were significantly decreased compared with those before surgery (P < 0.05). The levels of serum hs-CRP at 20 min and 24 hr after surgery were increased compared with those before surgery (P < 0.05). The level of serum hs-CRP at 14 d after surgery was significantly lower than that before surgery (P < 0.05). The correlation between SOD and hs-CRP was positive before surgery (r = 0.03, P < 0.001), but negative at 24 hr after surgery (r = -0.008, P < 0.001). The levels of serum SOD were significantly lower than median value (P < 0.05), while the Levels of serum hs-CRP were significantly higher than median value (P < 0.05). Conclusions: The levels of serum SOD and hs-CRP were significantly different before and after interventional treatment. The levels of serum SOD and hs-CRP can be used as indicators for the efficacy and prognosis of interventional treatment on type 2 diabetic patients with lower extremity vascular disease.

Mu, Yongxu; Yan, Ruiqiang; Hu, Xiaoyan; He, Junfeng; Liu, Haiyan; Li, Qiming

2015-01-01

187

Simple Stratification of Survival Using Bone Scan and Serum C-Reactive Protein in Prostate Cancer Patients with Metastases  

Microsoft Academic Search

Background: IL-6 has been reported to be a significant prognostic factor for prostate cancer and induces synthesis of C-reactive protein (CRP) by hepatocytes. The present study was undertaken to evaluate the clinical value of serum CRP in prostate cancer patients with metastases. Methods: The prognostic significance of serum CRP as well as tumor histology, extent of disease (EOD) on bone

Jun Nakashima; Eiji Kikuchi; Akira Miyajima; Ken Nakagawa; Mototsugu Oya; Takashi Ohigashi; Masaru Murai

2008-01-01

188

Immunosuppressive acidic protein serum levels in breast cancer patients in a reference to CA 15-3 levels  

Microsoft Academic Search

Immunosuppressive acidic protein (IAP) has been described as a tumor marker in a number of malignant diseases. To evaluate the clinical importance of IAP in breast cancer patients, IAP serum level was determined in 75 breast cancer patients, using single radial immunodiffusion. Serum samples were also tested for CA 15-3. Cut off value for IAP was determined according to IAP

Arnon D. Cohen; Yehuda Shoenfeld; Jacob Gopas; Yoram Cohen

1994-01-01

189

Changes in dietary protein intake has no effect on serum cystatin C levels independent of the glomerular filtration rate  

Microsoft Academic Search

Cystatin C is being considered as a replacement for serum creatinine in the estimation of the glomerular filtration rate (GFR); however, its plasma levels might be affected by factors other than the GFR, such as protein intake. We performed a post hoc analysis of the data in the Modification of Diet in Renal Disease study, in which we compared serum

Navdeep Tangri; Lesley A Stevens; Christopher H Schmid; Gerald J Beck; Tom Greene; Josef Coresh; Andrew S Levey

2011-01-01

190

A multiplex serum protein assay for determining the probability of colorectal cancer.  

PubMed

Our purpose is to develop a serum assay to determine an individual's probability of having colorectal cancer (CRC). We have discovered a protein panel yielding encouraging, clinically significant results. We evaluated 431 serum samples from donors screened for CRC by colonoscopy. We compared the concentration of seven proteins in individuals with CRC versus individuals found to be CRC free. The assay monitored a single peptide from each of seven proteins. Comparing CRC to normal samples in univariate two-sample t-tests, 6 of the 7 proteins yielded a p-value less than 0.01. Logistic regression was used to construct a model for determination of CRC probability. The model was fit on a randomly chosen training set of 321 samples. Using 6 of the 7 proteins (ORM1, GSN, C9, HABP2, SAA2, and C3) and a cut point of 0.4, an independent test set of 110 samples yielded a sensitivity of 93.75%, a specificity of 82.89% and a prevalence-adjusted negative predictive value (NPV) of 99.9775% for the assay. The results demonstrate that the assay has promise as a sensitive, non-invasive diagnostic test to provide individuals with an understanding of their own probability of having CRC. PMID:22957311

Brock, Randall; Xiong, Bob; Li, Lily; Vanbogelen, Ruth A; Christman, Lori

2012-01-01

191

Searching for early breast cancer biomarkers by serum protein profiling of pre-diagnostic serum; a nested case-control study  

PubMed Central

Background Serum protein profiles have been investigated frequently to discover early biomarkers for breast cancer. So far, these studies used biological samples collected at or after diagnosis. This may limit these studies' value in the search for cancer biomarkers because of the often advanced tumor stage, and consequently risk of reverse causality. We present for the first time pre-diagnostic serum protein profiles in relation to breast cancer, using the Prospect-EPIC (European Prospective Investigation into Cancer and nutrition) cohort. Methods In a nested case-control design we compared 68 women diagnosed with breast cancer within three years after enrollment, with 68 matched controls for differences in serum protein profiles. All samples were analyzed with SELDI-TOF MS (surface enhanced laser desorption/ionization time-of-flight mass spectrometry). In a subset of 20 case-control pairs, the serum proteome was identified and relatively quantified using isobaric Tags for Relative and Absolute Quantification (iTRAQ) and online two-dimensional nano-liquid chromatography coupled with tandem MS (2D-nanoLC-MS/MS). Results Two SELDI-TOF MS peaks with m/z 3323 and 8939, which probably represent doubly charged apolipoprotein C-I and C3a des-arginine anaphylatoxin (C3adesArg), were higher in pre-diagnostic breast cancer serum (p = 0.02 and p = 0.06, respectively). With 2D-nanoLC-MS/MS, afamin, apolipoprotein E and isoform 1 of inter-alpha trypsin inhibitor heavy chain H4 (ITIH4) were found to be higher in pre-diagnostic breast cancer (p < 0.05), while alpha-2-macroglobulin and ceruloplasmin were lower (p < 0.05). C3adesArg and ITIH4 have previously been related to the presence of symptomatic and/or mammographically detectable breast cancer. Conclusions We show that serum protein profiles are already altered up to three years before breast cancer detection. PMID:21871081

2011-01-01

192

Effect of Soy Protein on Serum Lipid Profile and Some Lipid-metabolizing Enzymes in Cholesterol Fed Rats  

Microsoft Academic Search

2 Abstract: The effect of soy protein on serum lipid profile and some lipid metabolizing enzymes in rats fed with cholesterol diets was examined in this study. Rats were subjected to feeding trial over a period of six weeks on formulated diets containing: 20% soy protein with 0% cholesterol (group A); 20% soy protein with 5% cholesterol (group B), 20%

E. Chukwu Onyeneke; Olarewaju M. Oluba; Olalekan Adeyemi; George E. Eriyamremu; Samuel I. Ojeaburu; Kayode E. Adebisi; Oyeyemi Adeyemi

2007-01-01

193

Uptake of proteins and degradation of human serum albumin by Plasmodium falciparum – infected human erythrocytes  

PubMed Central

Background Intraerythrocytic malaria parasites actively import obligate nutrients from serum and export proteins and lipids to erythrocyte cytoplasm and membrane. The import of macromolecules in the malaria parasite has been the subject of many debates. To understand the import of macromolecules by the parasite, we studied the uptake of proteins by Plasmodium falciparum infected human erythrocyte. Methods Proteins were biotin labelled individually, purified on a gel filtration column and added to uninfected and infected asynchronized culture. The uptake of these proteins by malaria parasites was determined by western blot analysis of parasite pellet and their different fractions using streptavidin-horseradish conjugate. To further confirm this import, we studied the uptake of 125I-labelled proteins by western blot analysis as well as used direct immunofluorescence method. Results Here we show that biotin labelled and radio-iodinated polypeptides of molecular sizes in the range of 45 to 206 kDa, when added in the culture medium, get direct access to the parasite membrane through a membrane network by by-passing the erythrocyte cytosol. The import of these polypeptides is ATP-dependent as sodium azide treatment blocks this uptake. We also show that malaria parasites have the ability to take up and degrade biotin labelled human serum albumin, which has been shown to be essential for the parasite growth. Conclusions These results can be used, as a basis to explore the role of human serum albumin in the intraerythrocytic development of parasites, and this in turn can be an important adjunct to the development of novel antimalarial drugs. PMID:12801422

El Tahir, Ahmed; Malhotra, Pawan; Chauhan, Virander S

2003-01-01

194

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

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

Microfluidic Electrochemical Immunoarray for Ultrasensitive Detection of Two Cancer Biomarker Proteins in Serum  

PubMed Central

A microfluidic electrochemical immunoassay system for multiplexed detection of protein cancer biomarkers was fabricated using a molded polydimethylsiloxane channel and routine machined parts interfaced with a pump and sample injector. Using off-line capture of analytes by heavily-enzyme-labeled 1 ?m superparamagnetic particle (MP)-antibody bioconjugates and capture antibodies attached to an 8-electrode measuring chip, simultaneous detection of cancer biomarker proteins prostate specific antigen (PSA) and interleukin-6 (IL-6) in serum was achieved at sub-pg mL?1 levels. MPs were conjugated with ~90,000 antibodies and ~200,000 horseradish peroxidase (HRP) labels to provide efficient off-line capture and high sensitivity. Measuring electrodes feature a layer of 5 nm glutathione-decorated gold nanoparticles to attach antibodies that capture MP-analyte bioconjugates. Detection limits of 0.23 pg mL?1 for PSA and 0.30 pg mL?1 for IL-6 were obtained in diluted serum mixtures. PSA and IL-6 biomarkers were measured in serum of prostate cancer patients in total assay time 1.15 h and sensor array results gave excellent correlation with standard enzyme-linked immunosorbent assays (ELISA). These microfluidic immunosensors employing nanostructured surfaces and off-line analyte capture with heavily-labeled paramagnetic particles hold great promise for accurate, sensitive multiplexed detection of diagnostic cancer biomarkers. PMID:21632234

Chikkaveeraiah, Bhaskara V.; Mani, Vigneshwaran; Patel, Vyomesh; Gutkind, J. Silvio; Rusling, James F.

2011-01-01

197

Folate-receptor 1 (FOLR1) protein is elevated in the serum of ovarian cancer patients  

PubMed Central

Objectives Ovarian cancer is the most lethal gynecological malignancy in North America. Although survival rates are high when the disease is diagnosed at an early stage, this decreases exponentially in late-stage diagnoses. As such, there is a need for novel early detection biomarkers. Through an integrated approach to ovarian cancer biomarker discovery that combines proteomics with transcriptomics and bioinformatics, our laboratory has identified folate-receptor 1 (FOLR1) and Dickkopf-related protein 3 (Dkk-3) as putative biomarkers. The objective of this study was to measure the levels of FOLR1 and Dkk-3 in the serum of patients with ovarian cancer, benign gynecological conditions and healthy women. Design and Methods FOLR1 and Dkk-3 were analyzed in serum of 100 ovarian cancer patients, 100 patients with benign gynecological conditions, and 100 healthy women using enzyme-linked immunosorbent assays (ELISAs). All specimens were analyzed in triplicate. Results FOLR1 was significantly elevated in the serum of ovarian cancer patients compared to serum of both healthy controls (p < 0.0001) and patients with benign gynecological conditions (p < 0.0001). Furthermore, FOLR1 was strongly correlated with CA125 as both were elevated in the serous histotype and in late-stage disease. FOLR1 did not outperform CA125 in receiver operating characteristic curve analysis and there was no significant complementarity between the two markers. Dkk-3 was not significantly different between the three serum cohorts and was not correlated with CA125. Conclusions FOLR1 is a new biomarker for ovarian cancer which correlates closely with CA125. The role of FOLR1 in the pathogenesis of ovarian cancer warrants further investigation. PMID:23528302

Leung, F.; Dimitromanolakis, A.; Kobayashi, Hiroshi; Diamandis, E.P.; Kulasingam, V.

2014-01-01

198

Pharmacokinetics of warfarin in rats: role of serum protein binding and tissue distribution  

SciTech Connect

The purpose of this study was to explore the role of serum protein binding and tissue distribution in the non-linear pharmacokinetics of warfarin in rats. The first phase of the research was an attempt to elucidate the causes of intersubject differences in serum protein binding of warfarin in rats. It was found that the distribution of S-warfarin between blood and liver, kidneys, muscle, or fatty tissue was non-linear. Based on the tissue distribution data obtained, a physiologically-based pharmacokinetic model was developed to describe the time course of S-warfarin concentrations in the serum and tissues of rats. The proposed model was able to display the dose-dependent pharmacokinetics of warfarin in rats. Namely a lower clearance and a smaller apparent volume of distribution with increasing dose, which appear to be due to the presence of capacity-limited, high-affinity binding sites for warfarin in various tissues. To determine if the binding of warfarin to the high-affinity binding sites in the liver of rats is reversible, concentrations of S-warfarin in the liver and serum of rats were monitored for a very long time after an intravenous injection of a 1 mg/kg dose. In another study in rats, non-radioactive warfarin was found to be able to displace tissue-bound C/sup 14/-warfarin which was administered about 200 hours before the i.v. injection of the non-radioactive warfarin, showing that the binding of warfarin to the high-affinity binding sites in the body is persistent and reversible.

Cheung, W.K.

1985-01-01

199

Interaction of Silver Nanoparticles with Serum Proteins Affects Their Antimicrobial Activity In Vivo  

PubMed Central

The emergence of multidrug-resistant bacteria is a global threat for human society. There exist recorded data that silver was used as an antimicrobial agent by the ancient Greeks and Romans during the 8th century. Silver nanoparticles (AgNPs) are of potential interest because of their effective antibacterial and antiviral activities, with minimal cytotoxic effects on the cells. However, very few reports have shown the usage of AgNPs for antibacterial therapy in vivo. In this study, we deciphered the importance of the chosen methods for synthesis and capping of AgNPs for their improved activity in vivo. The interaction of AgNPs with serum albumin has a significant effect on their antibacterial activity. It was observed that uncapped AgNPs exhibited no antibacterial activity in the presence of serum proteins, due to the interaction with bovine serum albumin (BSA), which was confirmed by UV-Vis spectroscopy. However, capped AgNPs [with citrate or poly(vinylpyrrolidone)] exhibited antibacterial properties due to minimized interactions with serum proteins. The damage in the bacterial membrane was assessed by flow cytometry, which also showed that only capped AgNPs exhibited antibacterial properties, even in the presence of BSA. In order to understand the in vivo relevance of the antibacterial activities of different AgNPs, a murine salmonellosis model was used. It was conclusively proved that AgNPs capped with citrate or PVP exhibited significant antibacterial activities in vivo against Salmonella infection compared to uncapped AgNPs. These results clearly demonstrate the importance of capping agents and the synthesis method for AgNPs in their use as antimicrobial agents for therapeutic purposes. PMID:23877702

Gnanadhas, Divya Prakash; Ben Thomas, Midhun; Thomas, Rony; Raichur, Ashok M.

2013-01-01

200

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

2015-06-01

201

Examination of serum pregnancy-associated plasma protein A clinical value in acute coronary syndrome prediction and monitoring  

PubMed Central

Introduction Chronic vascular inflammatory process promotes and intensifies all atherogenic events. The aim of this research was to estimate the clinical value of pregnancy-associated plasma protein A (PAPP-A) measurement associated with plaque destabilization and rupture in prediction and monitoring of acute coronary syndromes (ACS) as well as to assess the predictive value of this biomarker in comparison to traditional myocardial infarction (MI) risk markers. Material and methods The study included 119 patients in 2 investigated groups and one control group. PAPP-A assay was performed using manual ELISA kit, DRG. All other parameters were determined using automatic analyzers: Olympus and Dade Behring. Results A statistically significant difference between PAPP-A concentration median value was found in the investigated group MI individuals’ serum and control group individuals’ serum (11.42 ng/ml and 7.22 ng/ml respectively, p = 0.003). PAPP-A assay had the highest specificity (83.3%) and sensitivity (53.8%), and therefore the highest clinical value. In patients with clinically and laboratory confirmed MI we proved that PAPP-A serum level is a clinically useful biomarker in ACS prediction, better than C-reactive protein (hsCRP) and fibrinogen (FBG) level. Conclusions The highest diagnostic efficiency for ACS prediction was proved for simultaneous panel assays consisting of 2-3 parameters (PAPP-A – hsCRP, PAPP-A – FBG, PAPP-A – hsCRP – FBG), while PAPP-A itself does not show characteristics necessary for it to be used as a biomarker for MI dynamic monitoring. It is possible that prothrombotic component is mainly responsible for repeated major adverse cardiac events, more than inflammatory process. PMID:23515702

Rysz, Jacek; Paradowski, Marek

2013-01-01

202

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

203

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

204

Differential expression profiling of serum proteins and metabolites for biomarker discovery  

NASA Astrophysics Data System (ADS)

A liquid chromatography-mass spectrometry (LC-MS) proteomics and metabolomics platform is presented for quantitative differential expression analysis. Proteome profiles obtained from 1.5 [mu]L of human serum show ~5000 de-isotoped and quantifiable molecular ions. Approximately 1500 metabolites are observed from 100 [mu]L of serum. Quantification is based on reproducible sample preparation and linear signal intensity as a function of concentration. The platform is validated using human serum, but is generally applicable to all biological fluids and tissues. The median coefficient of variation (CV) for ~5000 proteomic and ~1500 metabolomic molecular ions is approximately 25%. For the case of C-reactive protein, results agree with quantification by immunoassay. The independent contributions of two sources of variance, namely sample preparation and LC-MS analysis, are respectively quantified as 20.4 and 15.1% for the proteome, and 19.5 and 13.5% for the metabolome, for median CV values. Furthermore, biological diversity for ~20 healthy individuals is estimated by measuring the variance of ~6500 proteomic and metabolomic molecular ions in sera for each sample; the median CV is 22.3% for the proteome and 16.7% for the metabolome. Finally, quantitative differential expression profiling is applied to a clinical study comparing healthy individuals and rheumatoid arthritis (RA) patients.

Roy, Sushmita Mimi; Anderle, Markus; Lin, Hua; Becker, Christopher H.

2004-11-01

205

Effects of periodontal therapy on C-reactive protein and HDL in serum of subjects with periodontitis  

PubMed Central

Objective To investigate the effects of nonsurgical periodontal therapy on levels of high-sensitivity C-reactive protein in the sera and its association with body mass index and high density lipoprotein in subjects with severe periodontitis. Methods Sera from 28 subjects (mean age: 34.36±6.24; 32% men) with severe periodontitis and 27 healthy controls (mean age: 33.18±6.42; 33% men) were collected prior to periodontal therapy. Blood samples were obtained from 23 subjects who completed therapy (9-12 months). Oral and systemic parameters such as the number of blood cells, glucose examination, lipid profile, and high-sensitivity C-reactive protein levels accessed by high-sensitivity immunonephelometry assay, were included. Results Before therapy, in the periodontitis group, the ratio of subjects with high-sensitivity C-reactive protein <0.3 mg/dL was statistically lower than in the control group (P<0.0216). After therapy, the ratio of subjects with high-sensitivity C-reactive protein <0.3 mg/dL was significantly higher (65.22%) (P<0.0339). The mean value for body mass index was statistically lower in subjects with high-sensitivity C-reactive protein <0.3 mg/dL (24.63±4.19), compared with those with high-sensitivity C-reactive protein >0.3 mg/dL (28.91±6.03) (P<0.0411). High density lipoprotein presented a mean value statistically higher after therapy (P<0.0027). Conclusion In systemically healthy subjects with periodontitis, periodontal therapy was associated with decreased levels of circulating high-sensitivity C-reactive protein and increase of high density lipoprotein in serum. The clinical trial was registered at http://www.clinicaltrials.gov.br/, No. RBR-24T799. PMID:24896165

Leite, Anne Carolina Eleutério; Carneiro, Valéria Martins de Araújo; Guimarães, Maria do Carmo Machado

2014-01-01

206

Speciation of trace elements in proteins in human and bovine serum by size exclusion chromatography and inductively coupled plasma-mass spectrometry with a magnetic sector mass spectrometer.  

PubMed

Proteins are separated by size exclusion chromatography while atomic ions from the inorganic elements are detected on-line by inductively coupled plasma-mass spectrometry. A double focusing mass analyzer provides very high sensitivity, low background, and sufficient spectral resolution to separate the atomic ions of interest from most polyatomic ions at the same nominal m/z value. The chromatograms show the distribution of the elements of interest between protein-bound and free fractions and provide the approximate molecular weights of those protein fractions that contain the elements monitored. The distribution of various elements, including V, Mo, Fe, Co, Mn, and lanthanides, in human or bovine serum samples are shown. Alkali metals and Tl are present primarily as free metal ions and are not bound to proteins. Inorganic elements spiked into the serum samples can be followed into various proteins. EDTA does not remove Fe, Pb, Sn, or Th from the proteins but does extract Mn from some proteins. Procedures for determining the effects of breaking disulfide linkages on the metal binding characteristics of proteins are also described. PMID:10550683

Wang, J; Houk, R S; Dreessen, D; Wiederin, D R

1999-10-01

207

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

208

Differential Effects of Corticosteroids on Serum Eosinophil Cationic Protein and Cytokine Production in Rhinovirus and Respiratory Syncytial Virus-Induced Acute Exacerbation of Childhood Asthma  

Microsoft Academic Search

Background: Little information is available on eosinophil activation and the cytokine profile in virus-induced acute exacerbation of bronchial asthma; therefore, we examined the effects of treatments that included systemic corticosteroids on serum eosinophil cationic protein (ECP) and 17 cytokines\\/chemokines in rhinovirus- and respiratory syncytial (RS) virus-induced acute exacerbation of childhood asthma. Methods: We measured the peripheral eosinophil count, as well

Masahiko Kato; Yoshiyuki Yamada; Kenichi Maruyama; Yasuhide Hayashi

2011-01-01

209

Embryo culture in teratological surveillance and serum proteins in development. Comprehensive fourteen year report, 1968-1982  

SciTech Connect

A testing procedure is being developed which may reduce the incidence of birth defects. The procedure uses in vitro cultures of whole rat embryos. Early studies on the nutrition of embryos involved attempts to culture whole chick embryos on protein-free chemically defined media. Media containing proteins from whole egg were superior. No single protein would support growth and development, at least two proteins were required. One protein was a transferrin, the second protein could be either ovalbumin or lipovitellin. To determine the path taken by nutrient proteins from medium to embryo, radioactive ovalbumin was prepared. The results showed that intact ovalbumin was taken up by the extra-embryonic yolk-sac and degraded to constituent amino acids within this structure. This observation was difficult to reconcile with the observed responses of the embryo to nutrient proteins. Consideration was given to yolk-sac function. When isolated yolk-sacs were incubated in Ringer's salt solution, they synthesized and secreted a distinct group of proteins. Yolk-sacs cultured on media containing various protein constituents synthesized serum proteins in relative amounts that were distinct for each medium. This suggested that the embryo responses to various culture media were mediated by changes in the relative synthesis of serum proteins. This hypothesis led to two lines of experimentation: what are the mechanisms regulating the relative synthesis of serum proteins by the yolk-sac; and do serum proteins actually provide signals of developmental significance. The first question led to studies with cultures of endodermal cells while the second question led to work on the development of a test for teratological surveillance. (ERB)

Klein, N.W.

1982-07-01

210

Synergy of serum and cerebrospinal fluid antibodies against axonal cytoskeletal proteins in patients with different neurological diseases.  

PubMed

Autoantibodies against different axonal cytoskeletal proteins [the light (NFL) and medium (NFM) subunit of neurofilament and tubulin (TUB)] in serum and cerebrospinal fluid may be generated in response to the release of cytoskeleton from damaged neurons. We studied the relationships among these autoantibodies. Paired cerebrospinal fluid (CSF) and serum samples were obtained from 47 multiple sclerosis (MS) patients, 14 patients with neurodegenerative diseases, 21 patients with various neurological diseases and 16 normal control subjects. Levels of antibodies against NFL, NFM and TUB were related to each other in CSF in all groups, whereas close association of anti-cytoskeletal antibodies in serum was found in the MS group only. A concordant spectrum of anti-cytoskeletal antibodies is present in serum of MS patients, unlike in other neurological patients. The synergy between the spectrum of anti-cytoskeletal antibodies in serum and CSF might be one of the immunological features typical for the MS patients. PMID:19445843

Fialová, L; Bartos, A; Soukupová, J; Svarcová, J; Ridzon, P; Malbohan, I

2009-01-01

211

The Prognostic Significance of Pretreatment Serum CEA Levels in Gastric Cancer: A Meta-Analysis Including 14651 Patients  

PubMed Central

Background Carcinoembryonic antigen (CEA) is commonly used as a serum tumor marker in clinical practice; however, its prognostic value for gastric cancer patients remains uncertain. This meta-analysis was performed to assess the prognostic value of CEA and investigate CEA as a tumor marker. Methods PubMed, EMBASE and other databases were searched for potentially eligible studies. Forty-one studies reporting the prognostic effect of pretreatment serum CEA expression in gastric cancer patients were selected. Data on 14651 eligible patients were retrieved for the meta-analysis. Based on the data extracted from the available literature, the hazard ratio (HR) and 95% confidence interval (CI) for an adverse prognosis were estimated for gastric cancer patients with elevated pretreatment serum levels of CEA (CEA+) relative to patients with normal pretreatment CEA levels (CEA-). Results The CEA+ patients had a significantly poorer prognosis than the CEA- patients in terms of overall survival (OS: HR 1.716, 95% CI 1.594 - 1.848, P< 0.001), disease-specific survival (DSS: HR 1.940, 95% CI 1.563 - 2.408, P< 0.001), and disease-free survival (DFS: HR 2.275, 95% CI 1.836 - 2.818, P< 0.001). Publication bias and an influence of different cut-off values were not observed (all P> 0.05). In the pooled analyses of multivariate-adjusted HRs, the results suggested that pretreatment serum CEA may be an independent prognostic factor in gastric cancer (OS: HR 1.681, 95% CI 1.425 - 1.982; DSS: HR 1.900, 95% CI 1.441 - 2.505; DFS: HR 2.579, 95% CI 1.935 - 3.436). Conclusion/Significance The meta-analysis based on the available literature supported the association of elevated pretreatment serum CEA levels with a poor prognosis for gastric cancer and a nearly doubled risk of mortality in gastric cancer patients. CEA may be an independent prognostic factor for gastric cancer patients and may aid in determining appropriate treatment which may preferentially benefit the CEA+ patients. PMID:25879931

Deng, Kai; Yang, Li; Hu, Bing; Wu, Hao; Zhu, Hong; Tang, Chengwei

2015-01-01

212

Peptide Mimicrying Between SARS Coronavirus Spike Protein and Human Proteins Reacts with SARS Patient Serum  

PubMed Central

Molecular mimicry, defined as similar structures shared by molecules from dissimilar genes or proteins, is a general strategy used by pathogens to infect host cells. Severe acute respiratory syndrome (SARS) is a new human respiratory infectious disease caused by SARS coronavirus (SARS-CoV). The spike (S) protein of SARS-CoV plays an important role in the virus entry into a cell. In this study, eleven synthetic peptides from the S protein were selected based on its sequence homology with human proteins. Two of the peptides D07 (residues 927–937) and D08 (residues 942–951) were recognized by the sera of SARS patients. Murine hyperimmune sera against these peptides bound to proteins of human lung epithelial cells A549. Another peptide D10 (residues 490–502) stimulated A549 to proliferate and secrete IL-8. The present results suggest that the selected S protein regions, which share sequence homology with human proteins, may play important roles in SARS-CoV infection. PMID:18320019

Hwa, K.-Y.; Lin, W. M.; Hou, Y.-I.; Yeh, T.-M.

2008-01-01

213

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

214

Validation processes of protein biomarkers in serum--a cross platform comparison.  

PubMed

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

215

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

216

The Anion Gap and Routine Serum Protein Measurements in Monoclonal Gammopathies  

PubMed Central

Summary Background and objectives An abnormal anion gap and an increased total protein and globulin are clues to the diagnosis of monoclonal gammopathy. We explored the utility of these markers in IgG, IgA, IgM, and free light chain monoclonal gammopathies. Design, Setting, Participants, & Measurements The anion gap, Na+ – (Cl– + HCO3–), corrected for hypoalbuminemia, was calculated in patients with monoclonal gammopathies. Exclusion criteria were serum calcium >10.5 mg/dl and/or creatinine >2 mg/dl. Results Among 287 patients, 242 remained after applying exclusion criteria (109 IgG, 64 IgA, 21 IgM, and 48 light chain); 36% of 242 patients required correction for hypoalbuminemia. The anion gap was decreased (<10) in 22% of IgG and increased (>15) in 31% of IgA monoclonal gammopathies. IgM did not affect the gap. In light chain gammopathies, the anion gap showed no consistent trend (15% increased, 17% decreased). Mean clonal IgG, IgA, and IgM concentrations were 10-fold higher than mean clonal free light chain concentrations in the respective monoclonal gammopathies (P < 0.001). These paraprotein level disparities were reflected in significantly increased mean serum total protein and globulin concentrations in IgG, IgA, and IgM versus free light chain monoclonal gammopathies, where mean total protein and globulin levels were within normal limits (P < 0.001). Conclusions The anion gap was significantly altered in IgG and IgA monoclonal gammopathies, but it was not a sensitive tool for suspecting the diagnosis. In light chain monoclonal gammopathies, the anion gap, total protein, and globulin did not provide reliable diagnostic clues. PMID:22157711

Joseph, Rosy E.; Gaughan, William J.; McBride, Laura; Bilotti, Elizabeth; McNeill, Ann; Schmidt, Linda; Schillen, Danielle; Siegel, David S.

2011-01-01

217

The combined effects of protein intake and resistance training on serum osteocalcin concentrations in strength and power athletes.  

PubMed

The purpose of the present investigation was to examine the combined effects of protein intake and resistance training on a blood marker of bone osteogenesis, serum osteocalcin, in Division III football players. Thirty-three resistance-trained football players (age = 20.4 +/- 1.8 years; height = 180.9 +/- 7.0 cm; body mass = 97.2 +/- 12.6 kg) were evaluated on their protein intake and subsequently underwent 10 weeks of periodized heavy resistance training during the off season. Subjects were then placed into 1 of the following 3 groups based on relative protein intake and were instructed to maintain their diets throughout the experimental period: (a) low protein intake (<1.2 g per kilogram of body mass), (b) moderate protein intake (1.21 to 1.90 g per kilogram of body mass), or (c) high protein intake (>1.91 g per kilogram of body mass). Blood sampling occurred prior to and following the 10-week resistance training period to determine resting serum osteocalcin concentrations. A significant main effect was observed following training such that only the group who consumed <1.2 g per kilogram of body mass of protein shown significant elevations in serum osteocalcin in response to resistance training (26.8 +/- 6.4 to 33.4 +/- 6.6 ng.ml(-1)). In addition, absolute protein intake was significantly correlated to serum osteocalcin concentrations (r = 0.39). The results of the present investigation demonstrated relationships between protein intake and serum osteocalcin concentrations. In addition, the osteocalcin response to short-term resistance training appeared to be affected by protein intake. PMID:18076269

Ratamess, Nicholas A; Hoffman, Jay R; Faigenbaum, Avery D; Mangine, Gerald T; Falvo, Michael J; Kang, Jie

2007-11-01

218

The human urinary proteome contains more than 1500 proteins, including a large proportion of membrane proteins  

PubMed Central

Background Urine is a desirable material for the diagnosis and classification of diseases because of the convenience of its collection in large amounts; however, all of the urinary proteome catalogs currently being generated have limitations in their depth and confidence of identification. Our laboratory has developed methods for the in-depth characterization of body fluids; these involve a linear ion trap-Fourier transform (LTQ-FT) and a linear ion trap-orbitrap (LTQ-Orbitrap) mass spectrometer. Here we applied these methods to the analysis of the human urinary proteome. Results We employed one-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis and reverse phase high-performance liquid chromatography for protein separation and fractionation. Fractionated proteins were digested in-gel or in-solution, and digests were analyzed with the LTQ-FT and LTQ-Orbitrap at parts per million accuracy and with two consecutive stages of mass spectrometric fragmentation. We identified 1543 proteins in urine obtained from ten healthy donors, while essentially eliminating false-positive identifications. Surprisingly, nearly half of the annotated proteins were membrane proteins according to Gene Ontology (GO) analysis. Furthermore, extracellular, lysosomal, and plasma membrane proteins were enriched in the urine compared with all GO entries. Plasma membrane proteins are probably present in urine by secretion in exosomes. Conclusion Our analysis provides a high-confidence set of proteins present in human urinary proteome and provides a useful reference for comparing datasets obtained using different methodologies. The urinary proteome is unexpectedly complex and may prove useful in biomarker discovery in the future. PMID:16948836

Adachi, Jun; Kumar, Chanchal; Zhang, Yanling; Olsen, Jesper V; Mann, Matthias

2006-01-01

219

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

220

A solvent system for delipidation of plasma or serum without protein precipitation  

Microsoft Academic Search

A technique has been developed which attains ih 30 minutes complete removal of triglyceride, cholesterol, phos- pholipid, and unesterified fatty acids from plasma without protein denaturation. Plasma is agitated at room temperature with a mixture of butanol and di-isopropyl ether in a 40:60 (v\\/v) ratio. The plasma proteins, including the apolipo- proteins, remain in solution in the aqueous phase, while

Bill E. Cham; Brian R. Knowlee

221

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

222

Structure and surface properties of the serum heat-induced protein aggregates isolated from heated skim milk  

Microsoft Academic Search

Serum heat-induced milk protein aggregates were isolated from heated skim milk. Protein analysis showed that the aggregates were essentially composed of whey protein and ?-casein linked mainly by disulfide bridges. The hydrodynamic diameter of the aggregates, as measured by dynamic light scattering (DLS), was about 70nm. Multi-angle DLS analysis and electron microscopy indicated that they were almost of spherical shape.

Karine Jean; Marie Renan; Marie-Hélène Famelart; Fanny Guyomarc’h

2006-01-01

223

Protein G binding to enriched serum immunoglobulin from nondomestic hoofstock species.  

PubMed

Quick and cost-effective serologic assays, such as those based on enzyme-linked immunosorbent assay (ELISA) technology, are useful for screening animal populations for infectious diseases. Recombinant protein G is described as an almost universal ELISA conjugate for the detection of antibodies from a wide range of animal species. However, there is limited data documenting the ability of protein G to bind immunoglobulin (Ig) from many captive and free-ranging nondomestic hoofstock (Order Artiodactyla, e.g., elk, antelope, bison). Protein G binding to Ig from 11 species within this taxonomic order (addax, antelope, bison, bontebok, elk, impala, kudu/nyala, muntjac, oryx, sheep, and white-tailed deer) and 2 control species (bovine and chicken) was assessed. A serum Ig enrichment protocol, using high-performance liquid chromatography (HPLC), was optimized in bovids (Bos taurus) and then applied to the other study species. Binding assays were performed by adding protein G to microtiter wells coated with titrated dilutions of enriched artiodactyl Ig. Optical densities were measured and binding curves generated. Differences in protein G binding were observed, both within and among species, as well as within taxonomic families. Significant intraspecies binding variation was observed for 7 species tested (antelope, oryx, sheep, muntjac, impala, bontebok, and addax). No statistically significant intraspecies differences in protein G binding were found for Ig from bison, elk, kudu/nyala, white-tailed deer, plus control species (cattle and chicken). Binding of protein G to Ig from impala, muntjac, and elk was statistically different from the positive control (cattle), with muntjac binding curves statistically comparable with the negative control (chicken). For the other 7 species tested, binding curves illustrated the ability of protein G to bind Ig as well as, or better than, the positive control. These findings expand the list of animal species whose Ig is capable of being detected using recombinant protein G, with the caveat that protein G does not bind Ig uniformly in closely related species. It is concluded that recombinant protein G conjugates may serve as useful reagents for serodiagnosis by ELISA in nondomestic hoofstock, although different assay interpretation algorithms and assay protocols may need to be developed on a per species basis for maximum diagnostic effectiveness. PMID:12735347

Kramsky, Joely A; Manning, Elizabeth J B; Collins, Michael T

2003-05-01

224

Altered phospholipid transfer protein gene expression and serum lipid profile by topotecan  

E-print Network

therapy [3]. The discovery of P-glycoprotein and its related proteins, encoded by the superfamily of ATP April 2010 Keywords: Topoisomerase I inhibitor Camptothecin Topotecan Phospholipid transfer protein High structural analogues including topotecan and irinotecan, are inhibitors of topoisomerase I. These drugs

Toledo, University of

225

Network motifs that recur across species, including gene regulatory and protein-protein interaction networks.  

PubMed

Cellular molecules interact in complex ways, giving rise to a cell's functional outcomes. Conscientious efforts have been made in recent years to better characterize these patterns of interactions. It has been learned that many of these interactions can be represented abstractly as a network and within a network there in many instances are network motifs. Network motifs are subgraphs that are statistically overrepresented within networks. To date, specific network motifs have been experimentally identified across various species and also within specific, intracellular networks; however, motifs that recur across species and major network types have not been systematically characterized. We reason that recurring network motifs could potentially have important implications and applications for toxicology and, in particular, toxicity testing. Therefore, the goal of this study was to determine the set of intracellular, network motifs found to recur across species of both gene regulatory and protein-protein interaction networks. We report the recurrence of 13 intracellular, network motifs across species. Ten recurring motifs were found across both protein-protein interaction networks and gene regulatory networks. The significant pair motif was found to recur only in gene regulatory networks. The diamond and one-way cycle reversible step motifs were found to recur only in protein-protein interaction networks. This study is the first formal review of recurring, intracellular network motifs across species. Within toxicology, combining our understanding of recurring motifs with mechanism and mode of action knowledge could result in more robust and efficient toxicity testing models. We are sure that our results will support research in applying network motifs to toxicity testing. PMID:24847787

Borotkanics, Robert; Lehmann, Harold

2015-04-01

226

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

227

Impact of Serum Proteins on MRI Contrast Agents: Cellular Binding and T2 relaxation.  

PubMed

Superparamagnetic iron oxide nanoparticles (SPIONs) used as MRI contrast agents or for theranostic applications encounter a complex mixture of extracellular proteins that adsorb on the SPION surface forming a protein corona. Our goal was to understand how cellular binding and T2 relaxation times are affected by this protein corona. Our studies focused on carboxymethyl dextran-modified SPIONs, chosen for their similarity to Resovist SPIONs used to detect liver lesions. Using a combination of fluorescence microscopy and flow cytometry, we find that the cellular binding of SPIONs to both macrophages and epithelial cells is significantly inhibited by serum proteins. To determine if this decreased binding is due to the iron oxide core or the carboxymethyl dextran surface coating, we functionalized polystyrene nanoparticles with a similar carboxymethyl dextran coating. We find a comparable decrease in cellular binding for the carboxymethyl dextran-polystyrene nanoparticles indicating that the carbohydrate surface modification is the key factor in SPION-cell interactions. NMR measurements showed that T2 relaxation times are not affected by corona formation. These results indicate that SPIONs have a decreased binding to cells under physiological conditions, possibly limiting their use in theranostic applications. We expect these results will be useful in the design of SPIONs for future diagnostic and therapeutic applications. PMID:25485101

Hill, Alexandra; Payne, Christine K

2014-01-01

228

Resistance of Neisseria meningitidis to Human Serum Depends on T and B Cell Stimulating Protein B.  

PubMed

The ability of the human bacterial pathogen Neisseria meningitidis to cause invasive disease depends on survival in the bloodstream via mechanisms to suppress complement activation. In this study, we show that prophage genes coding for T and B cell stimulating protein B (TspB), which is an immunoglobulin-binding protein, are essential for survival of N. meningitidis group B strain H44/76 in normal human serum (NHS). H44/76 carries three genes coding for TspB. Mutants having all tspB genes inactivated did not survive in >5% NHS or IgG-depleted NHS. TspB appeared to inhibit IgM-mediated activation of the classical complement pathway, since survival of the tspB triple knockout was the same as that of the parent strain or a complemented mutant when the classical pathway was inactivated by depleting NHS of C1q and was increased in IgM-depleted NHS. A mutant solely carrying tspB gene nmbh4476_0681 was as resistant as the parent strain, while mutants carrying only nmbh4476_0598 or nmbh4476_1698 were killed in ?5% NHS. The phenotype associated with TspB is formation of a matrix containing TspB, IgG, and DNA that envelopes aggregates of bacteria. Recombinant proteins corresponding to particular subdomains of TspB were found to have human IgG Fc?- and/or DNA-binding activity, but only TspB derivatives containing both domains formed large, biofilm-like aggregates when combined with purified IgG and DNA. Recognizing the role of TspB in serum resistance may lead to a better understanding of why strains that carry tspB genes are associated with invasive meningococcal disease. PMID:25583528

Müller, Maike G; Moe, Nina E; Richards, Phillip Q; Moe, Gregory R

2015-04-01

229

Acarbose treatment lowers generation and serum concentrations of the protein-bound solute p-cresol: a pilot study.  

PubMed

Several protein-bound uremic retention solutes (including p-cresol) originate from colonic bacterial fermentation of protein. Higher colonic availability of carbohydrates drives this process towards lower production of toxic metabolites. Small intestinal alpha-glucosidase inhibitors like Acarbose (Glucobay) enhance the amount of undigested carbohydrates reaching the colon. We studied the effect of Acarbose on generation and serum concentrations of p-cresol. Nine healthy volunteers (age 25 (22-36) years) with a creatinine clearance of 89.6 ml/min/1.73 m(2) (85.5-116.4) were treated with Acarbose for 3 weeks. Dose was gradually increased to reach 300 mg/day after 1 week. Blood sampling, 24-h urine and stool collections on 3 consecutive days were performed before and during the last days of the treatment period. p-Cresol generation was estimated from mean 24-h urinary elimination. Gastrointestinal side effects, if present, were mild to moderate. Serum concentrations of p-cresol declined significantly after Acarbose treatment (before: 1.14 mg/l (0.93-3.03); after: 1.11 mg/l (0.31-1.82); P=0.047). Urinary excretion of p-cresol, reflecting its colonic generation rate, was significantly lower after treatment (before: 29.93 mg/day (6.79-75.19); after: 10.54 mg/day (1.08-30.85); P=0.031). The fecal excretion of nitrogen increased after treatment (before: 1.04 g/day (0.47-2.29); after: 1.99 g/day (0.76-3.08); P=0.047). This pilot study suggests that Acarbose treatment lowers generation and serum concentrations of the protein-bound uremic solute p-cresol. Although further confirmation is warranted, the data may point to a novel treatment option for chronic kidney disease patients in view of the potential toxic effects of p-cresol and related substances. PMID:16688114

Evenepoel, P; Bammens, B; Verbeke, K; Vanrenterghem, Y

2006-07-01

230

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

231

Glycosaminoglycans in Human and Bovine Serum: Detection of Twenty-Four Heparan Sulfate and Chondroitin Sulfate Motifs Including a Novel Sialic Acid-modified Chondroitin Sulfate Linkage Hexasaccharide  

PubMed Central

Heterogeneous heparan sulfate and chondroitin sulfate glycosaminoglycan (GAG) polysaccharides are important components of blood circulation. Changes in GAG quantity and structure in blood have been indicated in cancers and other human diseases. However, GAG quantities and structures have not been fully characterized due to lack of robust and sensitive analytical tools. To develop such tools, we isolated GAGs from serum and plasma. We employed liquid chromatography (LC) for GAG quantification and LC/mass spectrometry (MS) for GAG structural analysis. Twenty-four heparan and chondroitin sulfate motifs were identified, including linkage hexasaccharides, repeating disaccharide compositions, reducing, and non-reducing end mono-, di-, tri-, and tetrasaccharide structures. Disaccharides were detectable at picomolar level without radiolabeling or derivitization, so only a few ml of human and fetal bovine serum was required for this study. The detection of different reducing end structures distinct from GAG linkage hexasaccharides revealed that free GAG chains generated by GAG degradation enzymes co-existed with proteoglycans in serum. In addition, a novel sialic acid-modified linkage hexasaccharide was found conjugated to bikunin, the most abundant serum proteoglycan. PMID:20657722

Lu, Hong; McDowell, Lynda M.; Studelska, Daniel R.; Zhang, Lijuan

2010-01-01

232

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. PMID:25624851

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

2013-01-01

233

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

PubMed

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. PMID:24722191

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

2014-05-01

234

Two-color, rolling-circle amplification on antibody microarrays for sensitive, multiplexed serum-protein measurements  

PubMed Central

The ability to conveniently and rapidly profile a diverse set of proteins has valuable applications. In a step toward further enabling such a capability, we developed the use of rolling-circle amplification (RCA) to measure the relative levels of proteins from two serum samples, labeled with biotin and digoxigenin, respectively, that have been captured on antibody microarrays. Two-color RCA produced fluorescence up to 30-fold higher than direct-labeling and indirect-detection methods using antibody microarrays prepared on both polyacrylamide-based hydrogels and nitrocellulose. Replicate RCA measurements of multiple proteins from sets of 24 serum samples were highly reproducible and accurate. In addition, RCA enabled reproducible measurements of distinct expression profiles from lower-abundance proteins that were not measurable using the other detection methods. Two-color RCA on antibody microarrays should allow the convenient acquisition of expression profiles from a great diversity of proteins for a variety of applications. PMID:15059261

2004-01-01

235

Protein Kinase C Signaling Complexes Include Metabolism and Transcription\\/ Translation-related Proteins  

Microsoft Academic Search

The serine\\/threonine kinase protein kinase C (PKC) has been shown to be a critical component in the heart's resistance to cell death following ischemic insult. Re- cent studies have indicated that PKC forms multi-pro- tein signaling complexes to accomplish signal transduc- tion in cardiac protection. Using two-dimensional electrophoresis (2DE), combined with matrix-assisted laser desorption ionization mass spectrometry (MS), the initial

Ricky D. Edmondson; Thomas M. Vondriska; Kelli J. Biederman; Jun Zhang; Richard C. Jones; Yuting Zheng; David L. Allen; Joanne X. Xiu; Ernest M. Cardwell; Michael R. Pisano; Peipei Ping

236

Serum Cholesterol  

Microsoft Academic Search

The relationship between low serum cholesterol and albumin levels were examined to determine if serum cholesterol would be useful in screening for protein-calorie malnutrition (PCM). Audits were conducted retrospectively of 80 randomly selected hospitalized patients over 60 years of age, on various forms of oral and parenteral nutrition. The study sample consisted of acutely ill patients with delayed oral intake

L. A. Zarny; L. H. Bernstein

1995-01-01

237

Ret finger protein inhibits muscle differentiation by modulating serum response factor and enhancer of polycomb1  

PubMed Central

Skeletal myogenesis is precisely regulated by multiple transcription factors. Previously, we demonstrated that enhancer of polycomb 1 (Epc1) induces skeletal muscle differentiation by potentiating serum response factor (SRF)-dependent muscle gene activation. Here, we report that an interacting partner of Epc1, ret finger protein (RFP), blocks skeletal muscle differentiation. Our findings show that RFP was highly expressed in skeletal muscles and was downregulated during myoblast differentiation. Forced expression of RFP delayed myoblast differentiation, whereas knockdown enhanced it. Epc1-induced enhancements of SRF-dependent multinucleation, transactivation of the skeletal ?-actin promoter, binding of SRF to the serum response element, and muscle-specific gene induction were blocked by RFP. RFP interfered with the physical interaction between Epc1 and SRF. Muscles from rfp knockout mice (Rfp?/?) mice were bigger than those from wild-type mice, and the expression of SRF-dependent muscle-specific genes was upregulated. Myotube formation and myoblast differentiation were enhanced in Rfp?/? mice. Taken together, our findings highlight RFP as a novel regulator of muscle differentiation that acts by modulating the expression of SRF-dependent skeletal muscle-specific genes. PMID:21637294

Kee, H J; Kim, J-R; Joung, H; Choe, N; Lee, S E; Eom, G H; Kim, J C; Geyer, S H; Jijiwa, M; Kato, T; Kawai, K; Weninger, W J; Seo, S B; Nam, K-I; Jeong, M H; Takahashi, M; Kook, H

2012-01-01

238

The relationship between serum sialic acid and high-sensitivity C-reactive protein with prehypertension  

PubMed Central

Background The aim of our study was to evaluate the serum concentration of sialic acid (SA) and high-sensitivity C-reactive protein (hs-CRP) in prehypertensive patients and the possible correlations between these 2 factors with blood pressure in such patients. Material/Methods We studied 61 prehypertensive patients, 70 hypertensive patients, and 50 controls with normal blood pressure. Lipid profile, hs-CRP, SA, and body mass index (BMI) were estimated in all groups. Associations between SA and hs-CRP and blood pressure were analyzed using multiple linear regressions. Results SA and hs-CRP levels were higher in the prehypertension group than that in the control group and were lower than that in the hypertension group. Multiple linear regression demonstrated that fasting glucose, BMI, SA, and hs-CRP correlated with systolic blood pressure and that low-density lipoprotein, BMI, SA, and hs-CRP correlated independently with diastolic pressure (P<0.05). Conclusions Our findings suggest that in prehypertension, there is an association between serum SA and hs-CRP levels and blood pressure. PMID:24694904

Jinghua, Li; Tie, Zhang; Ping, Wang; Yongtong, Cao

2014-01-01

239

Mutant p53 protein in serum could be used as a molecular marker in human breast cancer.  

PubMed

p53 wild-type is a tumor suppressor gene involved in DNA gene transcription or DNA repair mechanisms. When damage to DNA is unrepairable, p53 induces programmed cell death (apoptosis). The mutant p53 gene is the most frequent molecular alteration in human cancer, including breast cancer. Here, we analyzed the genetic alterations in p53 oncogene expression in 55 patients with breast cancer at different stages and in 8 normal women. We measured by ELISA assay the serum levels of p53 mutant protein and p53 antibodies. Immunohistochemistry and RT-PCR using specific p53 primers as well as mutation detection by DNA sequencing were also evaluated in breast tumor tissue. Serological p53 antibody analysis detected 0/8 (0%), 0/4 (0%) and 9/55 (16.36%) positive cases in normal women, in patients with benign breast disease and in breast carcinoma, respectively. We found positive p53 mutant in the sera of 0/8 (0.0%) normal women, 0/4 (0%) with benign breast disease and 29/55 (52.72%) with breast carcinoma. Immunohistochemistry evaluation was positive in 29/55 (52.73%) with mammary carcinoma and 0/4 (0%) with benign breast disease. A very good correlation between p53 mutant protein detected in serum and p53 accumulation by immunohistochemistry (83.3% positive in both assays) was found in this study. These data suggest that detection of mutated p53 could be a useful serological marker for diagnostic purposes. PMID:16525651

Balogh, G A; Mailo, D A; Corte, M M; Roncoroni, P; Nardi, H; Vincent, E; Martinez, D; Cafasso, M E; Frizza, A; Ponce, G; Vincent, E; Barutta, E; Lizarraga, P; Lizarraga, G; Monti, C; Paolillo, E; Vincent, R; Quatroquio, R; Grimi, C; Maturi, H; Aimale, M; Spinsanti, C; Montero, H; Santiago, J; Shulman, L; Rivadulla, M; Machiavelli, M; Salum, G; Cuevas, M A; Picolini, J; Gentili, A; Gentili, R; Mordoh, J

2006-04-01

240

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

241

Serum brain-derived neurotrophic factor levels in patients with major depression: Effects of antidepressants  

Microsoft Academic Search

This study tried to investigate the relationships between serum brain-derived neurotrophic factor (BDNF) protein levels and major depressive patients and discuss the effects of antidepressants on the serum BDNF protein levels. A total of 218 participants, including 111 patients with major depression (91 women) and 107 healthy controls (65 women), were recruited in this study. Serum BDNF protein levels were

Tiao-Lai Huang; Chien-Te Lee; Yu-Li Liu

2008-01-01

242

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

243

A Comparison of Blood Factor XII Autoactivation in Buffer, Protein Cocktail, Serum, and Plasma Solutions  

PubMed Central

Activation of blood plasma coagulation in vitro by contact with material surfaces is demonstrably dependent on plasma-volume-to-activator-surface-area ratio. The only plausible explanation consistent with current understanding of coagulation-cascade biochemistry is that procoagulant stimulus arising from the activation complex of the intrinsic pathway is dependent on activator surface area. And yet, it is herein shown that activation of the blood zymogen factor XII (Hageman factor, FXII) dissolved in buffer, protein cocktail, heat-denatured serum, and FXI deficient plasma does not exhibit activator surface-area dependence. Instead, a highly-variable burst of procoagulant-enzyme yield is measured that exhibits no measurable kinetics, sensitivity to mixing, or solution-temperature dependence. Thus, FXII activation in both buffer and protein-containing solutions does not exhibit characteristics of a biochemical reaction but rather appears to be a “mechanochemical” reaction induced by FXII molecule interactions with hydrophilic activator particles that do not formally adsorb blood proteins from solution. Results of this study strongly suggest that activator surface-area dependence observed in contact activation of plasma coagulation does not solely arise at the FXII activation step of the intrinsic pathway. PMID:23117212

Golas, Avantika; Yeh, Chyi-Huey Josh; Pitakjakpipop, Harit; Siedlecki, Christopher A.; Vogler, Erwin A.

2012-01-01

244

Effect of Langmuir monolayer of bovine serum albumin protein on the morphology of calcium carbonate  

E-print Network

Bovine serum albumin (BSA) Langmuir monolayer, as a model of biomineralization-associated proteins, was used to study its effect on regulated biomineralization of calcium carbonate. The effects of the BSA Langmuir monolayer and the concentration of the subphase solution on the nucleation and growth processes and morphology of the calcium carbonate crystal were investigated. The morphology and polymorphic phase of the resulting calcium carbonate crystals were characterized by scanning electron microscopy (SEM) and X-ray diffraction analysis (XRD). Moreover, the interaction mechanisms of the subphase solution with the BSA Langmuir monolayer were discussed. It was found that BSA Langmuir monolayer could be used as a template to successfully manipulate the polymorphic phase and crystal morphology of calcium carbonate and had obvious influence on the oriented crystallization and growth. The final morphology or aggregation mode of the calcite crystal was closely dependent on the concentration of calcium bicarbonate...

Xue, Zhong-Hui; Hu, Bin-Bin; Du, Zu-Liang; 10.1016/j.msec.2009.03.016

2011-01-01

245

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

246

Abundant expression of parathyroid hormone-related protein in primary rat aortic smooth muscle cells accompanies serum-induced proliferation.  

PubMed Central

Parathyroid hormone-related protein (PTHrP), which is responsible for producing hypercalcemia in patients with humoral hypercalcemia of malignancy, has recently been identified in several normal tissues. Because PTHrP, like parathyroid hormone (PTH), is known to exhibit vasodilatory properties, we investigated the expression and regulation of PTHrP mRNA in cultured rat aortic smooth muscle cells (SMC). We report here that PTHrP mRNA is expressed in SMC and is markedly induced by serum in a time- and concentration-dependent fashion. Addition of 10% fetal calf serum to serum-deprived, confluent cells, resulted in a marked induction of PTHrP mRNA by 2 h with a peak at 4-6 h. PTHrP was detected in SMC by immunocytochemistry and radioimmunoassay of conditioned medium, and was shown to be up-regulated within 24 h after the addition of serum. The serum induction of PTHrP mRNA was blocked by actinomycin D and by cycloheximide indicating the need for protein synthesis to evoke the serum effect on PTHrP gene transcription. In addition, treatment with dexamethasone, which has been previously shown to reduce the constitutive expression of PTHrP in human cancer cells, also blunted the serum induction of PTHrP mRNA in SMC. Treatment of quiescent cells with the serum mitogens platelet-derived growth factor or insulin-like growth factor-I had no effect on PTHrP, whereas the vasoactive peptides endothelin, norepinephrine and thrombin stimulated PTHrP expression. Exogenous addition of recombinant PTHrP-(1-141) had no significant effect on SMC DNA synthesis as measured by [3H]thymidine incorporation. In summary, the abundance of PTHrP mRNA and the characteristics of its regulation in SMC suggest a major role for PTHrP as a local modulator in vascular smooth muscle. Images PMID:1752945

Hongo, T; Kupfer, J; Enomoto, H; Sharifi, B; Giannella-Neto, D; Forrester, J S; Singer, F R; Goltzman, D; Hendy, G N; Pirola, C

1991-01-01

247

Relative Quantification of Serum Proteins from Pancreatic Ductal Adenocarcinoma Patients by Stable Isotope Dilution Liquid Chromatography-Mass Spectrometry  

PubMed Central

We report an innovative multiplexed liquid chromatography-multiple reaction monitoring/mass spectrometry (LC-MRM/MS)-based assay for rapidly measuring a large number of disease specific protein biomarkers in human serum. Furthermore, this approach uses stable isotope dilution methodology to reliably quantify candidate protein biomarkers. Human serum was diluted using a stable isotope labeled proteome (SILAP) standard prepared from the secretome of pancreatic cell lines, subjected to immunoaffinity removal of the most highly abundant proteins, trypsin digested, and analyzed by LC-MRM/MS. The method was found to be precise, linear, and specific for the relative quantification of 72 proteins when analyte response was normalized to the relevant internal standard (IS) from the SILAP. The method made it possible to determine statistically different concentrations for three proteins (cystatin M, IGF binding protein 7, and villin 2) in control and pancreatic cancer patient samples. This method proves the feasibility of using a SILAP standard in combination with stable isotope dilution LC-MRM/MS analysis of tryptic peptides to compare changes in the concentration of candidate protein biomarkers in human serum. PMID:22264027

Wehr, Angela Y.; Hwang, Wei-Ting; Blair, Ian A.; Yu, Kenneth H.

2012-01-01

248

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

249

Benefits and Limitations of Protein Hydrolysates as Components of Serum-Free Media for Animal Cell Culture Applications  

NASA Astrophysics Data System (ADS)

Increased understanding of influential factors for the cultivation of animal cells, combined with heightened regulatory concern over potential transmission of adventitious contaminants associated with serum and other animal-derived components, has elevated interest in using protein hydrolysates as serum replacements or nutrient supplements. This paper reviews the chemistry and biology of various hydrolysates derived from animal, plant and microbial sources. It provides specific examples of a beneficial selection of plant and yeast hydrolysates as ingredients of serum-free nutrient formulations for bioproduction applications of cultured mammalian and insect cells. Strategies for customizing and optimizing nutrients for specialized applications and general benefits and limitations of protein hydrolysates for biopharmaceutical production are also discussed.

Lobo-Alfonso, Juliet; Price, Paul; Jayme, David

250

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

251

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

252

Brief characterization of muskrat (Ondatra zibethicus) immunoglobulin G (IgG) separated from serum on protein A.  

PubMed

Muskrat (Ondatra zibethicus) immunoglobulin fraction was separated from whole serum by Protein A Sepharose chromatography. In serum electrophoresis, this fraction had a gamma motility; when electrophoresed on a polyacrylamide gel with sodium dodecyl sulfate it resolved into two protein bands of approximately 52 and 25 kilodaltons, respectively. These bands were consistent with molecular weights of known heavy and light chains of immunoglobulin G (IgG) in other closely related species. Furthermore, the putative muskrat immunoglobulins had a strong cross-reactivity with mouse IgG1, IgG3, and kappa chain in an enzyme-linked immunosorbent assay. We propose, that the proteins bound to the Protein A Sepharose represent muskrat immunoglobulins of the IgG class. PMID:9359072

Borucinska, J D; Ayoub, I; Garmendia, A E

1996-10-01

253

New insight into protein-nanomaterial interactions with UV-visible spectroscopy and chemometrics: human serum albumin and silver nanoparticles.  

PubMed

In recent years, great efforts have focused on the exploration and fabrication of protein nanoconjugates due to potential applications in many fields including bioanalytical science, biosensors, biocatalysis, biofuel cells and bio-based nanodevices. An important aspect of our understanding of protein nanoconjugates is to quantitatively understand how proteins interact with nanomaterials. In this report, human serum albumin (HSA) and citrate-coated silver nanoparticles (AgNPs) are selected as a case study of protein-nanomaterial interactions. UV-visible spectroscopy together with multivariate curve resolution by alternating least squares (MCR-ALS) algorithm is first exploited for the detailed study of AgNPs-HSA interactions. Introduction of the chemometrics tool allows extracting the kinetic profiles, spectra and distribution diagrams of two major absorbing pure species (AgNPs and AgNPs-HSA conjugate). These resolved profiles are then analysed to give the thermodynamic, kinetic and structural information of HSA binding to AgNPs. Transmission electron microscopy, circular dichroism spectroscopy and Fourier transform infrared spectroscopy are used to further characterize the complex system. Moreover, a sensitive spectroscopic biosensor for HSA is fabricated with the MCR-ALS resolved concentration of absorbing pure species. It is found that the linear range for the HSA nanosensor was from 1.9 nM to 45.0 nM with a detection limit of 0.9 nM. It is believed that the proposed method will play an important role in the fabrication and optimization of a robust nanobiosensor or cross-reactive sensors array for the detection and identification of biocomponents. PMID:24286103

Wang, Yong; Ni, Yongnian

2014-01-21

254

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...

255

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

256

A note on post partum utero vaginal prolapse in Gir cows: estimation of serum calcium, phosphorus, proteins, and cholesterol.  

PubMed

Reported are results obtained from determination of calcium, phosphorus, protein, and cholesterol in blood serum of 10 Gir cows with uterine prolapse. Particular reference is made to possible adverse effects of hypocalcaemia in conjunction with physiological stress resulting from pregnancy and parturition. PMID:2774812

Dhanotiya, R S; Srivastava, R K; Pandit, R K

1989-01-01

257

Quantization of bovine serum albumin by fluorescence enhancement effects and corresponding binding of macrocyclic host-protein assembly  

Microsoft Academic Search

The present paper reports the investigations on the spectroscopic behavior of the binary complexes of the dye aurintricarboxylic acid (ATA) with protein bovine serum albumin (BSA) and 18-crown 6 (CW) (ATA·BSA, ATA·CW) and the ternary complex ATA·CW·BSA by using UV–vis steady state and time resolved fluorescence spectroscopy. The primary aim of the work is to determine the protein (BSA) quantization

Munmun Bardhan; Tapas Misra; Tapan Ganguly

258

Preprocedural serum levels of C-reactive protein predict early complications and late restenosis after coronary angioplasty  

Microsoft Academic Search

OBJECTIVESWe sought to investigate whether early and late outcome after percutaneous transluminal coronary angioplasty (PTCA) could be predicted by baseline levels of acute-phase reactants.BACKGROUNDAlthough some risk factors for acute complications and restenosis have been identified, an accurate preprocedural risk stratification of patients undergoing PTCA is still lacking.METHODSLevels of C-reactive protein (CRP), serum amyloid A protein (SAA) and fibrinogen were measured

Antonino Buffon; Giovanna Liuzzo; Luigi M Biasucci; Patrizio Pasqualetti; Vito Ramazzotti; Antonio G Rebuzzi; Filippo Crea; Attilio Maseri

1999-01-01

259

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

260

Bioinformatics characterization of differential proteins in serum of mothers carrying Down syndrome fetuses: combining bioinformatics and ELISA  

PubMed Central

Introduction Characterization of novel proteins in maternal serum derived from mothers carrying Down syndrome (DS) fetuses. Material and methods Based on last comparative proteomic analysis, five significant differences of expressed proteins in serum from four groups have been confirmed by ELISA. DAVID and GeneGo MetaCore were used to bioinformatically analyze candidate protein markers. Results The serum levels of ceruloplasmin (CP) and complement factor B (CFB) were significantly increased in mother carried DS fetuses (346.5 ng/ml and 466.8 ng/ml vs. 248.6 ng/ml and 293.5 ng/ml, p< 0.05). Twenty-nine proteins were mainly categorized into binding, catalytic activity and enzyme regulator activity proteins, and their biological roles were involved in biological regulation, metabolic processes, cellular processes, and response to stimuli. The immune response alternative complement pathway was the most significant GeneGo Pathway related to DS. Conclusions These 29 proteins have relations with the development of Down syndrome, especially CP and CFB play more important roles. PMID:22661988

Yu, Bin; Zhang, Bin; Shi, Ye; Shao, Shi-he; Huang, Rui-ping; Yang, Yu-qi

2012-01-01

261

Total Protein Concentration and Diagnostic Test Results for Gray Wolf (Canis lupus) Serum using Nobuto Filter Paper Strips.  

PubMed

Nobuto filter paper strips are widely used for storing blood-serum samples, but the recovery of proteins from these strips following rehydration is unknown. Poor recovery of proteins could reduce the concentration of antibodies and antigens and reduce the sensitivity of diagnostic assays. We compared the protein concentration, and its association with test sensitivity, of eluted Nobuto strip samples with paired sera. We collected and froze serum from five gray wolves (Canis lupus) for 8 mo. When thawed, we used a spectrophotometer (absorbance 280 nm) to determine the serum protein concentration for paired sera and Nobuto eluates for each animal in 2-fold serial dilutions. Total protein concentration was similar for both sample storage methods (Nobuto eluates and control sera), except for the undiluted samples in which Nobuto eluates had higher total protein concentrations. Both sample storage methods appear to produce similar results using the SNAP® 4Dx® Test to detect antibodies against pathogens causing Lyme disease, anaplasmosis, and ehrlichiosis as well as antigen for canine heartworm disease. PMID:25574804

Jara, Rocío F; Sepúlveda, Carolina; Ip, Hon S; Samuel, Michael D

2015-04-01

262

Effect of Langmuir monolayer of bovine serum albumin protein on the morphology of calcium carbonate  

E-print Network

Bovine serum albumin (BSA) Langmuir monolayer, as a model of biomineralization-associated proteins, was used to study its effect on regulated biomineralization of calcium carbonate. The effects of the BSA Langmuir monolayer and the concentration of the subphase solution on the nucleation and growth processes and morphology of the calcium carbonate crystal were investigated. The morphology and polymorphic phase of the resulting calcium carbonate crystals were characterized by scanning electron microscopy (SEM) and X-ray diffraction analysis (XRD). Moreover, the interaction mechanisms of the subphase solution with the BSA Langmuir monolayer were discussed. It was found that BSA Langmuir monolayer could be used as a template to successfully manipulate the polymorphic phase and crystal morphology of calcium carbonate and had obvious influence on the oriented crystallization and growth. The final morphology or aggregation mode of the calcite crystal was closely dependent on the concentration of calcium bicarbonate solution. It is expected that this research would help to better understand the mechanism of biomineralization by revealing the interactions between protein matrices and crystallization of calcium carbonate crystal.

Zhong-Hui Xue; Shu-Xi Dai; Bin-Bin Hu; Zu-Liang Du

2011-06-14

263

The electrophoretical determination of serum protein fractions in lycopene treated experimental diabetic rats.  

PubMed

This study was planned to determine the effects of lycopene treatment on serum protein fractions in experimental diabetic rats. In order to induce diabetes in rats in the diabetes (D) and diabetes + lycopene (DL) groups, rats were given 45 mg/kg single-dose streptozotocin intraperitoneally. Lycopene (10 mg/kg/day dissolved in sunflower oil) was administered to the rats in the lycopene-only (L) and DL groups. Blood glucose levels and HbA1c% in DL group and diabetes group increased (p < 0.05) compared to control and L group. Total protein, albumin, ?1, ?2, and ? globulin fractions of diabetic and DL groups were lower than control and L groups (p < 0.05). D group had lowest gamma (?) globulin levels among other groups (p < 0.05). The ? globulin levels was slightly increased than diabetic groups (D and DL), but it was still lower than control and L groups (p < 0.05). The highest value of A/G ratio was observed in diabetic group. Similarly, the % level of A/G ratio of D group was higher than other groups. It was noted that the A/G ratio decreased and reached to control group levels after lycopene treatment. PMID:23712872

Yuksek, Veysel; Dede, Semiha; Ceylan, Ebubekir

2013-01-01

264

Association of Ca×PO4 product with levels of serum C-reactive protein in regular hemodialysis patients  

PubMed Central

Introduction: Numerous studies have attempted to identify risk factors for mortality and morbidity in maintenance hemodialysis patients. In this study we sought to examine the association of the levels of serum C-reactive protein (CRP) with value of Ca×PO4 product, in stable hemodialysis patients. Patients and Methods: Based on the severity of secondary hyperparathyroidism, patients being treated with oral active vitamin D3, calcium carbonate/Renagel tablets at various doses. Fasting serum 25-hydroxy vitamin D and intact serum parathormone and also serum blood urea nitrogen, CRP, calcium, phosphorus, alkaline phosphatase was measured. Results: A total of 41 patients, enrolled to the study. The mean patients’ age were 46(17.6) years. The value of serum CRP of patients was 8.6 (6.6) mg/l (median 6 mg/l). The value of Ca×PO4 product was 50.5(15.5) mg2/dl2 (median: 50 mg2/dl2). In this study, a significant inverse association between Ca×PO4 product and the age of the patients was seen. A significant positive correlation of logarithm of serum CRP with Ca×PO4 product was found. Conclusion: The result of this study, revealed the need to further attention to hyperphosphatemia in hemodialysis patients. PMID:25340109

Nasri, Hamid

2012-01-01

265

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

266

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

267

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

268

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

269

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

270

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

PubMed

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. 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

2014-02-01

271

Transcortin and vitamin D-binding protein levels in mouse serum.  

PubMed

The influence of age, sex and strain on the serum concentration of transcortin (corticosteroid-binding globulin) and vitamin D-binding protein (DBP) in mice was investigated. The effect of age was studied in two strains, C57BL/6JPfd and BALB/cmHeAPfd. The concentration of transcortin and DBP increased with age. In young animals the concentration of each protein showed a significant strain difference, which disappeared in older mice for DBP, but not for transcortin. In 7-day-old animals, no sex difference was observed for either protein, but in older animals a clear sex difference was found for transcortin. Adult males tended to have somewhat higher levels of DBP than adult females, but this difference was significant only on day 70. The variation in transcortin and DBP levels was further investigated in a large number of mouse strains. The DBP concentration did not markedly vary among strains (5.98-9.65 mumol/l in males and 5.08-8.85 mumol/l in females). Transcortin, however, showed marked strain variations, ranging from 0.72 to 2.06 mumol/l in males and from 1.02 to 4.55 mumol/l in females and there was a significant correlation (r = 0.66, n = 26, P less than 0.001) between the mean transcortin levels in males and females of different strains. Interstrain variation was much higher than intrastrain variation or variation among related strains, suggesting that the transcortin concentration is largely controlled by genetically determined factors. There was a significant correlation (r = 0.82, n = 9, P less than 0.01) between the mean corticosterone and transcortin concentrations (measured at 21.00 h).(ABSTRACT TRUNCATED AT 250 WORDS) PMID:3711757

Faict, D; De Moor, P; Bouillon, R; Heyns, W; Heiniger, H J; Corrow, D; Lesaffre, E

1986-05-01

272

Immunising with the transmembrane envelope proteins of different retroviruses including HIV-1: a comparative study.  

PubMed

The induction of neutralizing antibodies is a promising way to prevent retrovirus infections. Neutralizing antibodies are mainly directed against the envelope proteins, which consist of two molecules, the surface envelope (SU) protein and the transmembrane envelope (TM) protein. Antibodies broadly neutralizing the human immunodeficiencvy virus-1 (HIV-1) and binding to the TM protein gp41 of the virus have been isolated from infected individuals. Their epitopes are located in the membrane proximal external region (MPER). Since there are difficulties to induce such neutralizing antibodies as basis for an effective AIDS vaccine, we performed a comparative analysis immunising with the TM proteins of different viruses from the family Retroviridae. Both subfamilies, the Orthoretrovirinae and the Spumaretrovirinae were included. In this study, the TM proteins of three gammaretroviruses including (1) the porcine endogenous retrovirus (PERV), (2) the Koala retrovirus (KoRV), (3) the feline leukemia virus (FeLV), of two lentiviruses, HIV-1, HIV-2, and of two spumaviruses, the feline foamy virus (FFV) and the primate foamy virus (PFV) were used for immunisation. Whereas in all immunisation studies binding antibodies were induced, neutralizing antibodies were only found in the case of the gammaretroviruses. The induced antibodies were directed against the MPER and the fusion peptide proximal region (FPPR) of their TM proteins; however only the antibodies against the MPER were neutralizing. Most importantly, the epitopes in the MPER were localized in the same position as the epitopes of the antibodies broadly neutralizing HIV-1 in the TM protein gp41 of HIV-1, indicating that the MPER is an effective target for the neutralization of retroviruses. PMID:23249763

Denner, Joachim

2013-03-01

273

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

274

Serum and cerebrospinal fluid antibodies against myelin basic protein and their IgG subclass distribution in multiple sclerosis.  

PubMed Central

IgG class antibodies reactive with myelin basic protein (MBP) were determined by enzyme-linked immunosorbent assay (ELISA) in serum and cerebrospinal fluid (CSF) of 37 patients with multiple sclerosis and a control group of 32 patients with tension headache or psychoneurosis. Using standardised amounts of IgG from CSF and serum in ELISA, significantly higher mean antibody levels were found in CSF as well as in serum from the patients with multiple sclerosis. Ten (27%) of the multiple sclerosis CSF samples and 15 (41%) of the multiple sclerosis sera revealed anti MBP antibody levels exceeding 2 SD of the control group. Seven patients (19%) showed exclusive or higher levels of anti MBP antibodies in CSF, suggesting synthesis within the central nervous system. Analysis by ELISA for IgG subclasses of anti MBP antibodies revealed that they were restricted to IgG 1 in four patients and IgG 3 in one. PMID:2428940

García-Merino, A; Persson, M A; Ernerudh, J; Díaz-Gil, J J; Olsson, T

1986-01-01

275

Exposure to wireless phone emissions and serum beta-trace protein.  

PubMed

The lipocalin type of prostaglandin D synthase or beta-trace protein is synthesized in the choroid plexus, lepto-meninges and oligodendrocytes of the central nervous system and is secreted into the cerebrospinal fluid. beta-trace protein is the key enzyme in the synthesis of prostaglandin D2, an endogenous sleep-promoting neurohormone in the brain. Electromagnetic fields (EMF) in the radio frequency (RF) range have in some studies been associated with disturbed sleep. We studied the concentration of beta-trace protein in blood in relation to emissions from wireless phones. This study included 62 persons aged 18-30 years. The concentration of beta-trace protein decreased with increasing number of years of use of a wireless phone yielding a negative beta coefficient = -0.32, 95% confidence interval -0.60 to -0.04. Also cumulative use in hours gave a negative beta coefficient, although not statistically significant. Of the 62 persons, 40 participated in an experimental study with 30 min exposure to an 890-MHz GSM signal. No statistically significant change of beta-trace protein was found. In a similar study of the remaining 22 participitants with no exposure, beta-trace protein increased significantly over time, probably due to a relaxed situation. EMF emissions may down-regulate the synthesis of beta-trace protein. This mechanism might be involved in sleep disturbances reported in persons exposed to RF fields. The results must be interpreted with caution since use of mobile and cordless phones were self-reported. Awareness of exposure condition in the experimental study may have influenced beta-trace protein concentrations. PMID:20596612

Hardell, Lennart; Söderqvist, Fredrik; Carlberg, Michael; Zetterberg, Henrik; Mild, Kjell Hansson

2010-08-01

276

Influence of clotting time on the protein composition of serum samples based on LC-MS data.  

PubMed

Many large, disease-related biobanks of serum samples have been established prior to the widespread use of proteomics in biomarker research. These biobanks may contain relevant information about the disease process, response to therapy or patient classifications especially with respect to long-term follow-up that is otherwise very difficult to obtain based on newly initiated studies, particularly in the case of slowly developing diseases. An important parameter that may influence the composition of serum but that is often not exactly known is clotting time. We therefore investigated the influence of clotting time on the protein and peptide composition of serum by label-free and stable-isotope labeling techniques. The label-free analysis of trypsin-digested serum showed that the overall pattern of LC-MS data is not affected by clotting times varying from 2 to 8h. However, univariate and multivariate statistical analyses revealed that proteins that are directly involved in blood clot formation, such as the clotting-derived fibrinopeptides, change significantly. This is most easily detected in the supernatant of acid-precipitated, immunodepleted serum. Stable-isotope labeling techniques show that truncated or phosphorylated forms of fibrinopeptides A and B increase or decrease depending on clotting time. These patterns can be easily recognized and should be taken into consideration when analyzing LC-MS data using serum sample collections of which the clotting time is not known. Next to the fibrinopeptides, leucine-rich alpha-2-glycoprotein (P02750) was shown to be consistently decreased in samples with clotting times of more than 1h. For prospective studies, we recommend to let blood clot for at least 2h at room temperature using glass tubes with a separation gel and micronized silica to accelerate blood clotting. PMID:18996063

Govorukhina, Natalia I; de Vries, Marcel; Reijmers, Theo H; Horvatovich, Péter; van der Zee, Ate G J; Bischoff, Rainer

2009-05-01

277

Serum protein profile study of clinical samples using high performance liquid chromatography-laser induced fluorescence: case of cervical and oral cancers  

NASA Astrophysics Data System (ADS)

The serum protein profiles of normal subjects, patients diagnosed with cervical cancer, and oral cancer were recorded using High Performance Liquid Chromatography combined with Laser Induced Fluorescence detection (HPLC-LIF). Serum protein profiles of the above three classes were tested for establishing the ability of HPLC-LIF protein profiling technique for discrimination, using hard clustering and Fuzzy clustering methods. The clustering algorithms have quite successfully classified the profiles as belonging to normal, cancer of cervix, and oral cancer conditions.

Karemore, Gopal; Sujatha, .; Rai, Lavanya; Pai, Keerthilatha M.; Kartha, V. B.; Santhosh C., .

2009-02-01

278

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

279

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

PubMed

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

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

2014-08-01

280

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

281

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

282

Serum insulin-like growth factor (IGF)-I and IGF-binding proteins in lung cancer patients.  

PubMed

Many studies have shown that insulin-like growth factors (IGF-I & IGF-II) are implicated in the autocrine and paracrine growth of various tumors. Alterations in serum IGFs and IGF-binding proteins (IGFBPs) profiles have been reported in lung cancer. In this study, we measured serum levels of IGF-I and IGFBPs in 41 patients with lung cancer (small cell lung cancer, SCLC, 9; non-small cell lung cancer, NSCLC, 32) by radioimmunoassay and Western ligand blot (WLB). The serum IGF-I level in patients with lung cancer was significantly lower than in controls (207.9+/-62.6 vs 281.3+/-53.9 ng/mL, p<0.01). Patients with NSCLC showed significantly lower serum levels of IGF-I compared with SCLC patients (194.0+/-62.9 vs 258.4+/-27.8 ng/mL, p<0.01). Patients with squamous cell carcinoma tended to show lower serum levels of IGF-I than in those with adenocarcinoma (187.9+/-63.6 vs 215.9+/-59.5 ng/mL, p>0.05). The concentration of IGFBP-3 in lung cancer was 48% of that found in controls by WLB. The serum level of IGFBP-2 was markedly elevated in patients with lung cancer compared with controls (1303.7+/-618.0 vs 696.2+/-300.5, p<0.01). However, there was no significant difference between SCLC and NSCLC groups. This result showed that serum level of IGF-I/IGFBPs may be useful markers for diagnosing and identifying tumor types in lung cancer and further studies are needed. PMID:10485619

Lee, D Y; Kim, S J; Lee, Y C

1999-08-01

283

Postmortem serum nitrogen compounds and C-reactive protein levels with special regard to investigation of fatal hyperthermia.  

PubMed

The present study analyzed serum levels of urea nitrogen (UN), creatinine (Cr), and C-reactive protein (CRP), which are very stable during the early postmortem period, for investigation of the cause of death with special regard to hyperthermia (heat stroke) in serial medico-legal autopsy cases (n = 429), excluding fatal injury, intoxication, and fire fatality. In this series, mechanical asphyxiation, drowning, and sudden cardiac death cases (n = 56, n = 43, and n = 212, respectively) usually showed low levels within postmortem reference ranges for these serum markers, although UN and CRP levels were mildly elevated in cases of sudden cardiac death and cerebrovascular stroke. There were concomitant significant elevations in serum levels of UN (>50 mg/dL), Cr (>2 mg/dL), and CRP (>2 mg/dL) for chronic renal failure, gastrointestinal bleeding, pneumonia, and hypothermia (cold exposure). UN and CRP were especially high for chronic renal failure and pneumonia, respectively. However, hyperthermia cases showed an isolated elevation in the serum Cr level, suggesting an influence of systemic skeletal muscle damage. These serum markers may be practically useful for postmortem investigation of death due to hyperthermia (heat stroke), for which specific pathological and toxicological evidence may not be available. PMID:19291458

Maeda, Hitoshi; Zhu, Bao-Li; Bessho, Yasumori; Ishikawa, Takaki; Quan, Li; Michiue, Tomomi; Zhao, Dong; Li, Dong-Ri; Komatsu, Ayumi

2008-01-01

284

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

285

Maternal serum interleukin-6, C-reactive protein, and matrix metalloproteinase-9 concentrations as risk factors for preterm birth <32 weeks and adverse neonatal outcomes.  

PubMed

Elevated concentrations of interleukin-6 (IL-6), C-reactive protein (CRP), and matrix metalloproteinase-9 (MMP-9) in fetal and neonatal compartments have been associated with an increased risk for preterm birth (PTB) and/or neonatal morbidity. The purpose of this study was to determine if the maternal serum concentration of IL-6, CRP, and MMP-9 in women at risk for PTB, who are not in labor and have intact membranes, are associated with an increased risk for PTB <32 weeks and/or neonatal morbidity. Maternal serum samples collected from 475 patients enrolled in a multicenter randomized controlled trial of single versus weekly corticosteroids for women at increased risk for preterm delivery were assayed. Serum was collected at randomization (24 to 32 weeks' gestation). Maternal serum concentrations of IL-6, CRP, and MMP-9 were subsequently determined using enzyme-linked immunoassays. Multivariate logistic regression analysis was performed to explore the relationship between maternal serum concentrations of IL-6, CRP, and MMP-9 and PTB <32 weeks, respiratory distress syndrome (RDS), chronic lung disease (CLD), intraventricular hemorrhage (IVH), necrotizing enterocolitis (NEC), and any sepsis. Maternal serum concentrations of IL-6 and CRP, but not MMP-9, above the 90th percentile at the time of randomization were associated with PTB <32 weeks. In contrast, there was no significant relationship between RDS and NEC and the maternal serum concentration of IL-6, CRP, or MMP-9 (univariate analysis). The development of CLD was associated with a high (above 90th percentile) IL-6 and CRP in maternal serum, even after adjustment for gestational age (GA) at randomization and treatment group. However, when GA at delivery was added to the model, this finding was nonsignificant. Neonatal sepsis was more frequent in neonates born to mothers with a high maternal serum concentration of CRP (>90th percentile). However, there was no significant association after adjustment for GA at randomization and treatment group. Logistic regression analysis for each analyte indicated that high maternal serum concentrations of IL-6 and CRP, but not MMP-9, were associated with an increased risk of IVH (odds ratio [OR] 4.60, 95% confidence interval [CI] 1.86 to 10.68; OR 4.07, 95% CI 1.63 to 9.50) after adjusting for GA at randomization and treatment group. Most babies (25/30) had grade I IVH. When GA at delivery was included, elevated IL-6 remained significantly associated with IVH (OR 2.77, 95% CI 1.02 to 7.09). An elevated maternal serum concentration of IL-6 and CRP are risk factors for PTB <32 weeks and subsequent development of neonatal IVH. An elevated maternal serum IL-6 appears to confer additional risk for IVH even after adjusting for GA at delivery. PMID:20195952

Sorokin, Yoram; Romero, Roberto; Mele, Lisa; Wapner, Ronald J; Iams, Jay D; Dudley, Donald J; Spong, Catherine Y; Peaceman, Alan M; Leveno, Kenneth J; Harper, Margaret; Caritis, Steve N; Miodovnik, Menachem; Mercer, Brian M; Thorp, John M; O'Sullivan, Mary Jo; Ramin, Susan M; Carpenter, Marshall W; Rouse, Dwight J; Sibai, Baha

2010-09-01

286

Serum-induced translocation of mitogen-activated protein kinase to the cell surface ruffling membrane and the nucleus  

PubMed Central

The mitogen-activated protein (MAP) kinase signal transduction pathway represents an important mechanism by which growth factors regulate cell function. Targets of the MAP kinase pathway are located within several cellular compartments. Signal transduction therefore requires the localization of MAP kinase in each sub-cellular compartment that contains physiologically relevant substrates. Here, we show that serum treatment causes the translocation of two human MAP kinase isoforms, p40mapk and p41mapk, from the cytosol into the nucleus. In addition, we report that p41mapk (but not p40mapk) is localized at the cell surface ruffling membrane in serum-treated cells. To investigate whether the protein kinase activity of MAP kinase is required for serum-induced redistribution within the cell, we constructed mutated kinase-negative forms of p40mapk and p41mapk. The kinase-negative MAP kinases were not observed to localize to the cell surface ruffling membrane. In contrast, the kinase-negative MAP kinases were observed to be translocated to the nucleus. Intrinsic MAP kinase activity is therefore required only for localization at the cell surface and is not required for transport into the nucleus. Together, these data demonstrate that the pattern of serum-induced redistribution of p40mapk is different from p41mapk. Thus, in addition to common targets of signal transduction, it is possible that these MAP kinase isoforms may differentially regulate targets located in distinct sub-cellular compartments. PMID:8394846

1993-01-01

287

Pathogenic autoantibodies from patients with lupus nephritis cause reduced tyrosine phosphorylation of podocyte proteins, including tubulin  

PubMed Central

Introduction The tertiary structure of normal podocytes prevents protein from leaking into urine. Patients with lupus nephritis (LN) develop proteinuria, and kidney biopsies from these patients display a number of podocyte abnormalities including retraction of podocyte processes. Autoantibodies have been shown to deposit in the kidneys of patients and mice with LN and are believed to play a key role in causing renal inflammation and dysfunction. The objective of this research was to study the effects of IgG antibodies from patients with LN on cultured human podocytes. Methods We exposed a human podocyte cell line to heat-inactivated (HI) plasma and purified polyclonal IgG from the following groups of subjects; patients with LN, patients with lupus without nephritis, patients with rheumatoid arthritis and healthy controls. We measured expression and intracellular distribution of podocyte-specific proteins and global phosphorylation of tyrosine. We then used mass spectrometry to identify the major protein targets of this phosphorylation. Results HI LN plasma did not alter expression or cellular distribution of podocyte-specific proteins but caused a significant reduction in podocyte protein tyrosine phosphorylation compared with plasma from healthy controls (p=0.0008). This result was replicated using purified IgG but was not seen with plasma from rheumatoid arthritis or non-renal lupus patients. The dominant tyrosine phosphorylated protein in podocytes was 55?kDa in size and was identified as tubulin. Conclusions Since tubulin is an important component of podocyte major processes, these results suggest that autoantibodies from LN patients may exert an important pathogenic effect by dephosphorylation of this protein in podocytes. PMID:25396061

Manson, Jessica J; Mills, Kevin; Jury, Elizabeth; Mason, Lesley; D'Cruz, David P; Ni, Lan; Saleem, Moin; Mathieson, Peter; Isenberg, David; Rahman, Anisur

2014-01-01

288

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

289

The cyclic-AMP receptor protein (CRP) regulon in Aggregatibacter actinomycetemcomitans includes leukotoxin  

PubMed Central

The cyclic-AMP receptor protein (CRP) acts as a global regulatory protein among bacteria. Here, the CRP regulon has been defined in Aggregatibacter actinomycetemcomitans using microarray analysis of A. actinomycetemcomitans strain JP2 wild type cells compared to an isogenic crp deletion mutant. Genes whose expression levels changed at least 2-fold with p ? 0.05 were considered significant. Of the 300 genes identified as being CRP-regulated, 139 were CRP-activated, including leukotoxin, with the remaining being CRP-repressed. The 300 genes represent 14.2% of ORFs probed which is significantly higher than what has been reported for CRP regulons in other bacteria. If the CRP-regulated genes are put into 17 functional classes, all 17 categories had at least 1 CRP-regulated gene. Several functional categories, mainly transport and binding proteins and energy metabolism proteins, were disproportionately represented in the CRP-regulated subset of genes relative to their overall representation in the genome. This is similar to the patterns seen in other bacteria. Finally, quantitative RT-PCR was used to show that the leukotoxin RNA levels were repressed 16-fold in the CRP mutant indicating that CRP activates leukotoxin transcription. However, this regulation appears to be acting through another regulatory protein since the leukotoxin promoter, unlike ~129 other promoters of CRP-regulated genes, does not have a match to the consensus CRP binding site. Several candidate genes for this intermediary transcription factor have been identified in the CRP-regulon. PMID:21575705

Feuerbacher, Leigh A.; Burgum, Alex; Kolodrubetz, David

2011-01-01

290

Comparing serum responses to acute feedings of an extensively hydrolyzed whey protein concentrate versus a native whey protein concentrate in rats: a metabolomics approach.  

PubMed

We examined how gavage feeding extensively hydrolyzed whey protein (WPH) versus a native whey protein concentrate (WPC) transiently affected serum biochemical profiles in rodents. Male Wistar rats (250-300 g) were 8 h fasted and subsequently fed isonitrogenous amounts of WPH or WPC, or remained unfed (control). Animals were sacrificed 15 min, 30 min, and 60 min post-gavage for serum extraction, and serum was analyzed using untargeted global metabolic profiling via gas chromatography/mass spectrometry (MS) and liquid chromatography/MS/MS platforms. We detected 333 serum metabolites amongst the experimental and control groups. Both WPH and WPC generally increased amino acids (1.2-2.8-fold), branched-chain amino acids (1.2-1.7-fold), and serum di- and oligo-peptides (1.1-2.7-fold) over the 60 min time course compared with control (q < 0.05). However, WPH increased lysine (false discovery rate using a q-value <0.05) and tended to increase isoleucine and valine 15 min post-feeding (q < 0.10) as well as aspartylleucine 30 min post-feeding compared with WPC (q < 0.05). While both protein sources led to a dramatic increase in free fatty acids compared with control (up to 6-fold increases, q < 0.05), WPH also uniquely resulted in a 30 min post-feeding elevation in free fatty acids compared with WPC (q < 0.05), an effect which may be due to the robust 30 min postprandial increase in epinephrine in the WPH cohort. These data provide a unique postprandial time-course perspective on how WPH versus WPC feedings affect circulating biochemicals and will guide future research comparing these 2 protein sources. PMID:24476471

Roberts, Michael D; Cruthirds, Clayton L; Lockwood, Christopher M; Pappan, Kirk; Childs, Thomas E; Company, Joseph M; Brown, Jacob D; Toedebusch, Ryan G; Booth, Frank W

2014-02-01

291

A fluorescence-based high throughput assay for the determination of small molecule–human serum albumin protein binding  

PubMed Central

Herein, we describe the development of a fluorescence-based high throughput assay to determine the small molecule binding towards human serum albumin (HSA). This innovative competition assay is based on the use of a novel fluorescent small molecule Red Mega 500 with unique spectroscopic and binding properties. The commercially available probe displays a large fluorescence intensity difference between the protein-bound and protein-unbound state. The competition of small molecules for HSA binding in the presence of probe resulted in low fluorescence intensities. The assay was evaluated with the LOPAC small molecule library of 1280 compounds identifying known high protein binders. The small molecule competition of HSA–Red Mega 500 binding was saturable at higher compound concentrations and exhibited IC50 values between 3–24 ?M. The compound affinity towards HSA was confirmed by isothermal titration calorimetry indicating that the new protein binding assay is a valid high throughput assay to determine plasma protein binding. PMID:24390461

McCallum, Megan M.; Pawlak, Alan J.; Shadrick, William R.; Simeonov, Anton; Jadhav, Ajit; Yasgar, Adam; Maloney, David J.; Arnold, Leggy A.

2014-01-01

292

Comparison of Measured GFR, Serum Creatinine, Cystatin C, and Beta-Trace Protein to Predict ESRD in African Americans With Hypertensive CKD  

PubMed Central

Background Identification of persons with chronic kidney disease (CKD) who are at highest risk to progress to end stage renal disease (ESRD) is necessary to reduce the burden of kidney failure. The relative utility of traditional markers of kidney function, including estimated glomerular filtration rate (GFR) and serum creatinine, and emerging markers of kidney function, including cystatin C and beta-trace protein (BTP), to predict ESRD and mortality has yet to be established. Study Design Randomized clinical trial followed by an observational cohort study. Setting & Participants 865 African American individuals with hypertensive CKD enrolled in a clinical trial of two levels of blood pressure control and three different antihypertensive drugs as initial therapy and subsequently followed by an observational cohort study. Predictors Quintile of measured GFR (mGFR) by iothalamate clearance, serum creatinine, serum creatinine-based estimated GFR (eGFRSCr), cystatin C, and BTP. Outcomes and Measurements Incidence of ESRD and mortality. Results A total of 246 participants reached ESRD over a median follow-up of 102 months. The incidence rate of ESRD was higher with higher quintiles of each marker. The association between higher BTP and ESRD was stronger than those for the other markers, including mGFR. All the markers remained significantly associated with ESRD after adjustment for mGFR and relevant covariates (all p<0.05), with BTP retaining the strongest association (HR for highest versus lowest quintile, 5.7; 95% CI, 2.2-14.9). Associations with the combined endpoint of ESRD or mortality (n=390) were weaker, but remained significant for cystatin C (p=0.05) and BTP (p=0.004). Limitations The ability of these markers to predict ESRD and mortality in other racial and ethnic groups and among individuals with CKD due to other causes is unknown. Conclusions Plasma BTP and cystatin C may be useful adjuncts to serum creatinine and mGFR in evaluating risk for progression of kidney disease. PMID:21944667

Bhavsar, Nrupen A.; Appel, Lawrence J.; Kusek, John W.; Contreras, Gabriel; Bakris, George; Coresh, Josef; Astor, Brad C.

2011-01-01

293

Change in serum proteome during allogeneic hematopoietic stem cell transplantation and clinical significance of serum C-reactive protein and haptoglobin  

PubMed Central

Successful hematopoietic stem cell transplantation (HSCT) involves the restoration of hematopoietic function after engraftment, arising from the differentiation and proliferation of hematopoietic stem cells. Several factors could influence the course of allogeneic-HSCT (allo-HSCT). Therefore, knowledge of serum proteome changes during the allo-HSCT period might increase the efficacy of diagnosis and disease prevention efforts. This study conducted proteomic analyses to find proteins that were significantly altered in response to allo-HSCT. Sera from five representative patients who underwent allo-HSCT were analyzed by 2-dimensional gel electrophoresis and liquid chromatography tandem mass spectrometry, and were measured on a weekly basis before and after allo-HSCT in additional 78 patients. Fourteen protein spots showing changes in expression were further examined, and most proteins were identified as acute phase proteins (APPs). Studies of 78 additional patients confirmed that C-reactive protein (CRP) and haptoglobin undergo expression changes during allo-HSCT and thus may have the potential to serve as representative markers of clinical events after allo-HSCT. Maximal CRP level affected the development of major transplant-related complications (MTCs) and other problems such as fever of unknown origin. Particularly, an increase in CRP level 21 days after allo-HSCT was found to be an independent risk factor for MTC. Maximal haptoglobin and haptoglobin level 14 days after allo-HSCT were predictive of relapses in underlying hematologic disease. Our results indicated that CRP and haptoglobin were significantly expressed during allo-HSCT, and suggest that their level can be monitored after allo-HSCT to assess the risks of early transplant-related complications and relapse. PMID:20716902

Ryu, Joohyun; Lee, Se Ryeon; Park, Sung Goo; Kang, Sunghyun

2010-01-01

294

Clara Cell Secretory Protein in Tracheobronchial Aspirates and Umbilical Cord Serum of Extremely Premature Infants with Systemic Inflammation  

Microsoft Academic Search

Background: A systemic fetal inflammatory response, reflected by chorioamnionitis with funisitis, is a risk factor for bronchopulmonary dysplasia. Clara cell secretory protein (CC10), a product of pulmonary Clara cells, has anti-inflammatory properties. Local down-regulation of CC10 has been associated with inflammatory lung disease. Increased serum levels of CC10 can indicate injury to alveolar-capillary integrity. Objective: We hypothesized that extremely premature

Wolfgang Thomas; Silvia Seidenspinner; Maria Chmielnicka-Kopaczyk; Alexander Marx; Johannes Wirbelauer; Marta Szymankiewicz; Christian P. Speer

2010-01-01

295

Serum cartilage oligomeric matrix protein and clinical signs and symptoms of potential pre-radiographic hip and knee pathology  

Microsoft Academic Search

Objective To examine the cross-sectional relationship between serum cartilage oligomeric matrix protein (COMP) and hip and knee clinical signs and symptoms in a sample of adults without radiographic hip or knee osteoarthritis (OA).Design A total of 145 persons with available sera and no evidence of radiographic hip or knee OA (Kellgren–Lawrence grade 0) were randomly selected from the Caucasian participants

A. D. Dragomir; V. B. Kraus; J. B. Renner; G. Luta; A. Clark; V. Vilim; M. C. Hochberg; C. G. Helmick; J. M. Jordan

2002-01-01

296

Growth Factors and Steroid Mediated Regulation of Cytoskeletal Protein Expression in Serum-Deprived Primary Astrocyte Cultures  

Microsoft Academic Search

In this research we aimed to investigate the interactions between growth factors (GFs) and dexamethasone (DEX) on cytoskeletal\\u000a proteins GFAP and vimentin (VIM) expression under different experimental conditions. Condition I: 24 h pretreatment with bFGF,\\u000a subsequent 72 h switching in serum-free medium (SFM) and final addition of GFs, alone or by two in the last 24 h, after a\\u000a prolonged (60 h) DEX treatment.

Vincenzo Bramanti; Daniela Bronzi; Daniele Tomassoni; Antonino Costa; Giuseppina Raciti; Marcello Avitabile; Francesco Amenta; Roberto Avola

2008-01-01

297

Serum amyloid A induces IL-8 secretion through a G protein-coupled receptor, FPRL1/LXA4R.  

PubMed

Host response to injury and infection is accompanied by a rapid rise in the blood of acute-phase proteins such as serum amyloid A (SAA). Although SAA has been used as a marker for inflammatory diseases, its role in the modulation of inflammation and immunity has not been defined. Human neutrophils respond to SAA with secretion of the proinflammatory cytokines interleukin 8 (IL-8) and, to a lesser extent, tumor necrosis factor alpha (TNF-alpha). The induction of IL-8 secretion by SAA involves both transcription and translation and correlates with activation of nuclear factor kappaB (NF-kappaB). The proximal signaling events induced by SAA include mobilization of intracellular Ca(2+) and activation of the mitogen-activated protein kinases ERK1/2 and p38, both required for the induced IL-8 secretion. Pertussis toxin effectively blocks SAA-induced IL-8 secretion indicating involvement of a Gi-coupled receptor. Overexpression of FPRL1/LXA4R in HeLa cells results in a significant increase of the expression of NF-kappaB and IL-8 luciferase reporters by SAA, and an antibody against the N-terminal domain of FPRL1/LXA4R inhibits IL-8 secretion. Lipoxin A4, which binds to FPRL1/LXA4R specifically, decreases SAA-induced IL-8 secretion significantly. Collectively, these results indicate that the cytokine-like property of SAA is manifested through activation of the Gi-coupled FPRL1/LXA4R, which has been known to mediate the anti-inflammatory effects of lipoxin A4. The ability of FPRL1/LXA4R to mediate 2 drastically different and opposite functions suggests that it plays a role in the modulation of inflammatory and immune responses. PMID:12393391

He, Rong; Sang, Hairong; Ye, Richard D

2003-02-15

298

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

299

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

300

Stromal cells from human long-term marrow cultures, but not cultured marrow fibroblasts, phagocytose horse serum constituents: studies with a monoclonal antibody that reacts with a species-specific epitope common to multiple horse serum proteins.  

PubMed

This report describes an IgG1 mouse monoclonal antibody derived after immunization of mice with washed stromal cells from human, long-term bone marrow cultures. The antigen recognized by the antibody (BMS-1) is a carbohydrate-containing prosthetic group that is common to and specific for multiple horse serum proteins. These proteins are avidly ingested by stromal cells and concentrated in endocytic vesicles. Cultured smooth muscle cells took up the horse proteins in a similar manner to marrow stromal cells while cultured marrow fibroblasts, endothelial cells, and hepatoma cells did not. These data indicate that marrow stromal cells specifically accumulate horse serum proteins which might partially explain the horse serum requirement for long-term marrow culture maintenance. The data also suggest further similarities between marrow stromal and smooth muscle cells and additional differences between marrow fibroblasts and marrow stromal cells. PMID:3780891

Charbord, P; Tippens, D; Wight, T S; Gown, A M; Singer, J W

1987-01-01

301

Association of pretreatment serum interferon ? inducible protein 10 levels with sustained virological response to peginterferon plus ribavirin therapy in genotype 1 infected patients with chronic hepatitis C  

PubMed Central

Background Increased serum and intrahepatic interferon ? inducible protein 10 (IP?10) levels in patients with chronic hepatitis C (CHC) have been described. Aim To analyse the possible association of serum IP?10 levels with different outcomes to antiviral therapy. Patients A total of 137 CHC patients treated with peginterferon plus ribavirin. Methods Serum IP?10 levels were determined by enzyme linked immunosorbent assay before therapy, after 12?weeks of treatment, and 24?weeks after cessation of therapy. Variables significantly associated with a sustained virological response (SVR) on univariate analysis were included in a multivariate logistic regression model. Results Pretreatment serum IP?10 levels in patients with SVR were significantly lower than in non?responders (NR) (332.4 (222.1) v 476.8 (305.3)?pg/ml, respectively; p?=?0.004). Serum IP?10 concentrations significantly decreased in patients with SVR (pretreatment: 332.4 (222.1)?pg/ml; post?treatment: 170.2 (140.1)?pg/ml; p<0.001) but not in NR (pretreatment: 476.8 (305.3)?pg/ml; post treatment: 387.3 (268.1)?pg/ml; p?=?0.06). By multivariate analysis, non?1 genotype (odds ratio (OR) 3.5 (95% confidence interval (CI) 1.1–10.4); p?=?0.003) and low viral load at baseline (OR 0.34 (95% CI 0.14–0.79); p?=?0.01) were independent predictors of SVR in all patients. When multivariate analysis was restricted to patients with genotype 1, only baseline viral load (OR 0.38 (95% CI 0.155–0.96); p?=?0.04) and pretreatment serum IP?10 levels (OR 0.99 (95% CI 0.996–0.999); p?=?0.03) were identified as predictive factors of SVR. Conclusion Pretreatment serum IP?10 behaves as a predictive factor of SVR to peginterferon plus ribavirin therapy in genotype 1 infected patients. PMID:16150856

Diago, M; Castellano, G; García?Samaniego, J; Pérez, C; Fernández, I; Romero, M; Iacono, O L; García?Monzón, C

2006-01-01

302

Serum concentrations of IGF-I, estradiol-17?, testosterone, and relative amounts of IGF binding proteins (IGFBP) in growing boars, barrows, and gilts1,2  

Microsoft Academic Search

The purpose of this research was to de- termine whether serum concentrations of steroids, IGF- I, and relative amounts of serum IGF-binding proteins (IGFBP) differ in growing boars (n = 11), barrows (n = 11), and gilts (n = 12) from 70 to 140 d of age. Pigs of similar age and weight were housed in pens of three or

J. A. Clapper; T. M. Clark; L. A. Rempel

303

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

304

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

305

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

306

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. PMID:25684985

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

307

Fluctuations of Serum Neuron Specific Enolase and Protein S-100B Concentrations in Relation to the Use of Shunt during Carotid Endarterectomy  

PubMed Central

Objective To evaluate the changes in serum neuron specific enolase and protein S-100B, after carotid endarterectomy performed using the conventional technique with routine shunting and patch closure, or eversion technique without the use of shunt. Materials and Methods Prospective non-randomized study included 43 patients with severe (>80%) carotid stenosis undergoing carotid endarterectomy in regional anesthesia. Patients were divided into two groups: conventional endarterectomy with routine use of shunt and Dacron patch (csCEA group) and eversion endarterectomy without the use of shunt (eCEA group). Protein S-100B and NSE concentrations were measured from peripheral blood before carotid clamping, after declamping and 24 hours after surgery. Results Neurologic examination and brain CT findings on the first postoperative day did not differ from preoperative controls in any patients. In csCEA group, NSE concentrations decreased after declamping (P<0.01), and 24 hours after surgery (P<0.01), while in the eCEA group NSE values slightly increased (P=ns), accounting for a significant difference between groups on the first postoperative day (P=0.006). In both groups S-100B concentrations significantly increased after declamping (P<0.05), returning to near pre-clamp values 24 hours after surgery (P=ns). Sub-group analysis revealed significant decline of serum NSE concentrations in asymptomatic patients shunted during surgery after declamping (P<0.05) and 24 hours after surgery (P<0.01), while no significant changes were noted in non-shunted patients (P=ns). Decrease of NSE serum levels was also found in symptomatic patients operated with the use of shunt on the first postoperative day (P<0.05). Significant increase in NSE serum levels was recorded in non-shunted symptomatic patients 24 hours after surgery (P<0.05). Conclusion Variations of NSE concentrations seemed to be influenced by cerebral perfusion alterations, while protein S-100B values were unaffected by shunting strategy. Routine shunting during surgery for symptomatic carotid stenosis may have the potential to prevent postoperative increase of serum NSE levels, a potential marker of brain injury. PMID:25859683

Dragas, Marko; Koncar, Igor; Opacic, Dragan; Ilic, Nikola; Maksimovic, Zivan; Markovic, Miroslav; Ercegovac, Marko; Simic, Tatjana; Pljesa-Ercegovac, Marija; Davidovic, Lazar

2015-01-01

308

Combined inflammatory and metabolic defects reflected by reduced serum protein levels in patients with Buruli ulcer disease.  

PubMed

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

Phillips, Richard O; Sarfo, Fred S; Landier, Jordi; Oldenburg, Reid; Frimpong, Michael; Wansbrough-Jones, Mark; Abass, Kabiru; Thompson, William; Forson, Mark; Fontanet, Arnaud; Niang, Fatoumata; Demangel, Caroline

2014-04-01

309

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

310

Lower total serum protein, albumin, and beta- and gamma-globulin in major and treatment-resistant depression: Effects of antidepressant treatments  

Microsoft Academic Search

Strong evidence has recently been reported that major depression is accompanied by an acute phase response (APR), characterized by elevated levels of positive acute phase proteins (APPs) and decreased levels of negative APPs. The APR is also reflected in lowered total serum protein (TSP) and specific changes in the major electrophoretically separated protein fractions. The present study examined pretreatment and

Fran Van Hunsel; Annick Wauters; Eric Vandoolaeghe; Hugo Neels; Paul Demedts; Michael Maes

1996-01-01

311

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

PubMed

Biologic reactivity to orthopedic implant debris mediates long-term clinical performance of total joint arthroplasty implants. However, the reasons that some facets of implant debris (e.g., particle size, shape, base material, etc.) are more pro-inflammatory remain controversial. 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 proteinfilm adsorption, and adsorbed protein-film-dependent cytokine release using equal surface areas of different sized cobalt-chromium alloy (CoCr-alloy) particles and in vitro challenge of human macrophages (THP-1 and human primary). Smaller (5 ?m diameter) versus larger (70 ?m diameter) 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 a difference in cytokine expression in which albumin-coated particles induced more TNF-? release and IgG-coated particles induced more IL-1? release from human monocytes/macrophages. In these preliminary in vitro studies, we have 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, these findings suggest that adsorbed protein differences on different-sized particles of the same material may be a contributing mechanism by which certain particles induce different reactivities. PMID:24941408

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

2014-01-01

312

Embryo culture in teratological surveillance and serum proteins in development. Final technical report  

SciTech Connect

An overview of the authors research into teratogenesis of blood serum of patients on medication or rats injected with drugs is presented. In addition studies concerning the role of methionine in the developing fetus is given. 68 refs.

Klein, N.W.

1986-01-01

313

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

314

Developmental regulation of insulin-like growth factor binding protein-2 in chick embryo serum and vitreous humor.  

PubMed

The chick embryo is a useful vertebrate model for studying developmental embryogenesis. Insulin-like growth factor I (IGF-I), a potent mitogen, is thought to contribute to the general growth of the embryo as an endocrine factor, and as a paracrine factor to the development of the early embryo and of specific organs such as the eye. Recent data suggest that a family of at least six IGF binding proteins (IGFBPs) complex IGF-I and modulate its biological actions. In the present study, we examine the expression of IGFBPs in chicken serum and vitreous humor at different stages of embryonic development, and compare it with that of IGF-I. As determined by ligand blotting, the predominant IGFBP in chick serum and vitreous humor between embryonic days 4 and 22 (E4-E22) is a 30 kDa IGFBP. This IGFBP was specifically immunoprecipitated by a polyclonal antiserum raised against rat IGFBP-2, the predominant IGFBP in fetal human and rat serum. Although IGFBP-2 is present in both chick fluids at all times examined, serum IGFBP-2 increased progressively between E10-E22, whereas vitreous IGFBP-2 was highest during eye organogenesis (E4-E8). This suggests that vitreous IGFBP-2 is synthesized locally. Like serum IGFBP-2, levels of immunoreactive IGF-I in serum are higher in the second week of embryogenesis than the first. Despite this correlation, changes in IGFBP-2 do not appear to be regulated by IGF-I: (a) serum IGF-I decreases after day 15, whereas IGFBP-2 levels remain stable until hatching; (b) vitreous IGF-I, like serum IGF-I, is higher in the second week of embryogenesis, whereas vitreous IGFBP-2 is highest in the first week; (c) embryos cultured ex ovo express IGFBP-2 at E15-E19, although they lack the normal mid-embryogenesis surge in IGF-I. We conclude that vitreous IGFBP-2 is synthesized locally in the eye, and that the expression of IGFBP-2 in chick embryos is not directly regulated by IGF-I. PMID:7505461

Yang, Y W; Brown, D R; Robcis, H L; Rechler, M M; de Pablo, F

1993-10-20

315

The quantification of oxygen toxicity by the technique of cellulose acetate electrophoresis of rat serum proteins  

E-print Network

by cardiac puncture before and after exposure periods and serum was aspirated from the clotted and centrifuged samples. Serum samples were subjected to electrophoresis on cellulose acetate and examined for qualitative and quantitative changes... be of importance in oxygen toxicity studies and in the treatment of newborns with hyaline membrane disease. Although the process of hyalinization is not well understood, work done by Phillips, Wyte, Weeks and Soloway (116) and by Fujikura (63) indicated a...

Barker, Marcia Wagner

1979-01-01

316

Alexithymia and its relationships with C-reactive protein and serum lipid levels among drug naïve adult outpatients with major depression  

Microsoft Academic Search

Several studies have investigated the relationship between C-reactive protein (CRP) and serum lipid levels in Major Depression (MD), but no study has, to date, evaluated the impact of alexithymia on these parameters. Therefore, the aim of the present cross-sectional study was to evaluate the relationship between alexithymia, suicide risk, C-reactive protein (CRP) and serum lipid levels in adult outpatients suffering

Domenico De Berardis; Nicola Serroni; Daniela Campanella; Alessandro Carano; Francesco Gambi; Alessandro Valchera; Chiara Conti; Gianna Sepede; Marco Scali; Mario Fulcheri; Rosa Maria Salerno; Filippo Maria Ferro

2008-01-01

317

Enzyme-linked immunosorbent assay for detection of Plasmodium falciparum histidine-rich protein 2 in blood, plasma, and serum.  

PubMed

Microscopy, the gold standard for the detection and quantification of malaria parasites in blood, is in many aspects deficient for this purpose. The method is poorly reproducible and can be inaccurate because Plasmodium falciparum parasites sequester for a portion of each asexual cycle. Due to these deficiencies, biomarkers such as P. falciparum histidine-rich protein 2 (PfHRP2) are increasingly being used. In this study, we evaluated the use of a commercial PfHRP2 enzyme-linked immunosorbent assay (ELISA) kit with some procedural modifications. We determined the linear range of the assay, including the lower limits of detection and quantitation, using recombinant PfHRP2 (rPfHRP2). In 10 repeat experiments, the linear range of optical densities (ODs) at 450 to 650 nm was from 0.05 +/- 0.002 to 2.28 +/- 0.042, corresponding to 3.91 to 250 ng/ml of rPfHRP2. The coefficient of variation (CV) at each target concentration ranged from 1.93 to 8.07%. Using cultured parasites, we confirmed the linear range of ODs as well as the association between the PfHRP2 ELISA results and the microscopic parasite densities. For whole-blood samples spiked with cultured, washed, ring-stage-infected red blood cells (iRBCs), the linear range was 11.7 to 750 iRBCs/microl, with CVs of 0.29 to 7.56%. The same spiked samples evaluated by microscopists had similar sensitivities, but the CVs were unacceptably high (20.7 to 161.6%). Stock rPfHRP2 was stable through four freeze-thaw cycles (P < 0.05; paired t test). When different patient sample types at different concentrations within the linear range of the assay are compared, the recoveries of PfHRP2 from blood and serum were within +/-20%, whereas the recoveries from plasma ranged between +35 and -41%. We conclude that PfHRP2 ELISA using whole-blood and serum samples is a suitable adjunct to microscopy and could ultimately benefit malaria intervention trials. PMID:18367583

Kifude, Carolyne M; Rajasekariah, Halli G; Sullivan, David J; Stewart, V Ann; Angov, Evelina; Martin, Samuel K; Diggs, Carter L; Waitumbi, John N

2008-06-01

318

Value of serum pregnancy-associated plasma protein A for predicting cardiovascular events among patients presenting with cardiac chest pain  

PubMed Central

Background: Pregnancy-associated plasma protein A (PAPP-A) has been suggested as a candidate marker for the identification of unstable plaques in coronary arteries. We assessed the value of PAPP-A for predicting short-term cardiovascular events in a large cohort of patients presenting with cardiac chest pain. Methods: We included consecutive patients who presented to a teaching hospital in Germany with chest pain of cardiac origin confirmed by coronary angiography. We analyzed PAPP-A levels from serum samples drawn within 30 minutes after arrival in the emergency department or in the catheterization laboratory. Patients were followed for 90 days or until death for major adverse cardiovascular events, defined as the combined outcome of stent thrombosis, myocardial (re)infarction, ischemic stroke or cardiovascular-related death. Results: A total of 2568 patients (mean age [± standard deviation (SD)] 68 ± 11 years; 74% male) presented with cardiac chest pain: 55% had stable angina and 45% had acute coronary syndrome. The PAPP-A levels ranged from 4 to 2154 mIU/L (median 14.0 mIU/L, interquartile range 9.3–25.2 mIU/L). Major adverse cardiovascular events occurred in 203 patients (7.9%). The mean PAPP-A level was higher among patients who had an event than among those who did not (62 ± 156 v. 21 ± 23 mIU/L, p < 0.001). In a multivariable analysis, PAPP-A remained a significant independent predictor of the primary outcome within 90 days (hazard ratio per 1 SD increase in PAPP-A level 1.96, 95% confidence interval 1.74–2.21). The optimal prognostic cutoff value was a PAPP-A level of 34.6 mIU/L. Interpretation: Higher levels of serum PAPP-A were independently associated with an increased short-term risk of cardiovascular events in patients presenting with cardiac chest pain. Further studies are required to validate the use of PAPP-A in routine clinical practice to predict future cardiovascular events. PMID:23509133

von Haehling, Stephan; Doehner, Wolfram; Jankowska, Ewa A.; Ponikowski, Piotr; Stellos, Konstantinos; Puntmann, Valentina O.; Nagel, Eike; Anker, Stefan D.; Gawaz, Meinrad; Bigalke, Boris

2013-01-01

319

Serum levels of S100B and NSE proteins in Alzheimer's disease patients  

PubMed Central

Background Alzheimer's disease is the most common dementia in the elderly, and the potential of peripheral biochemical markers as complementary tools in the neuropsychiatric evaluation of these patients has claimed further attention. Methods We evaluated serum levels of S100B and neuron-specific enolase (NSE) in 54 mild, moderate and severe Alzheimer's disease (AD) patients and in 66 community-dwelling elderly. AD patients met the probable NINCDS-ADRDA criteria. Severity of dementia was ascertained by the Clinical Dementia Rating (CDR) scale, cognitive function by the Mini Mental State Examination (MMSE), and neuroimage findings with magnetic resonance imaging. Serum was obtained from all individuals and frozen at -70°C until analysis. Results By comparing both groups, serum S100B levels were lower in AD group, while serum NSE levels were the same both groups. In AD patients, S100B levels were positively correlated with CDR scores (rho = 0.269; p = 0.049) and negatively correlated with MMSE scores (rho = -0.33; P = 0.048). NSE levels decreased in AD patients with higher levels of brain atrophy. Conclusions The findings suggest that serum levels of S100B may be a marker for brain functional condition and serum NSE levels may be a marker for morphological status in AD. PMID:20105309

2010-01-01

320

Effect of 5-hydroxytryptamine on protein synthesis in gastrointestinal and other tissues and on serum gastrin concentrations in rats.  

PubMed Central

1 The effect of 5-hydroxytryptamine (5-HT) on protein synthesis in the gastrointestinal tissues as well as in the liver, heart and brain was studied by measuring [3H]-leucine incorporation into total tissue protein in vivo in rats. 2 A single injection of 5-HT (10 mg/kg) produced a marked inhibition (45 to 65%) in protein synthesis in the stomach (oxyntic gland area), intestine (jejunum + ileum), colon and brain, but not in the liver and heart. 3 Dose- and time-dependent studies of 5-HT-induced changes in protein synthesis in the stomach of fed rats revealed that the maximal inhibition of about 65% occurred 1 h after a dose of 12.5 mg/kg. 4 Animals deprived of food for 24 h differed from their fed counterparts only in the degree of responsiveness, in that a greater inhibition (75%) of [3H]-leucine incorporation into total protein of the stomach was observed 30 min after 5-HT injection. 5 Pretreatment of animals with methysergide (0.25 mg/kg) essentially abolished the 5-HT-mediated inhibition of protein synthesis in the stomach. 6 Serum gastrin concentration in fasted animals remained slightly elevated during the initial period of 5-HT treatment (up to 1 h). 7 The results demonstrate that in certain tissues, 5-HT markedly inhibits protein synthesis. The inhibition in protein synthesis in the gastrointestinal tract cannot be attributed to changes in the concentration of gastrin. PMID:465872

Nandi Majumdar, A. P.; Nakhla, A. M.

1979-01-01

321

Effect of protein dose on serum glucose and insulin response to sugars13  

Microsoft Academic Search

fasting decreased with increasing protein dose. Protein-containing meals produced significantly lower (p < 0.01) areas than the protein-free meal and the relationship between blood glucose area and protein dose was significant (p < 0.001). Protein-containing meals produced significantly greater (p < 0.0 1) insulin areas compared with the protein-free meal. However, no differences in insulin areas among the protein-containing meals

Gene A Spiller; Christopher D Jensen; Thomas S Pattison; Carol S Chuck; James H Whittam; James Scala

322

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. 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

2014-01-01

323

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

324

Surface shaving as a versatile tool to profile global interactions between human serum proteins and the Staphylococcus aureus cell surface.  

PubMed

The human commensal bacterium Staphylococcus aureus is renowned as a causative agent of severe invasive diseases. Upon entering the bloodstream, S. aureus can infect almost every tissue and organ system in the human body. To withstand insults from the immune system upon invasion, several immune-evasive mechanisms have evolved in S. aureus, such as complement inhibition by secreted proteins and IgG-binding by surface-exposed protein A. While it is generally accepted that S. aureus cells bind a range of host factors for various purposes, no global analyses to profile staphylococcal host factor binding have so far been performed. Therefore, we explored the possibility to profile the binding of human serum proteins to S. aureus cells by "surface shaving" with trypsin and subsequent MS analysis of liberated peptides. This resulted in the identification of several components of the complement system, the platelet factor 4 and the isoform 1 of the inter-?-trypsin inhibitor heavy chain H4 on the staphylococcal cell surface. We conclude that surface shaving is a versatile tool to profile global interactions between human serum proteins and the S. aureus cell surface. PMID:21674804

Dreisbach, Annette; van der Kooi-Pol, Magdalena M; Otto, Andreas; Gronau, Katrin; Bonarius, Hendrik P J; Westra, Hans; Groen, Herman; Becher, Dörte; Hecker, Michael; van Dijl, Jan M

2011-07-01

325

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

326

The cellular uptake of meta-tetra(hydroxyphenyl)chlorin entrapped in organically modified silica nanoparticles is mediated by serum proteins  

NASA Astrophysics Data System (ADS)

Nanosized objects made of various materials are gaining increasing attention as promising vehicles for the delivery of therapeutic and diagnostic agents for cancer. Photodynamic therapy (PDT) appears to offer a very attractive opportunity to implement drug delivery systems since no release of the sensitizer is needed to obtain the therapeutic effect and the design of the nanovehicle should be much easier. The aim of our study was to investigate the use of organic-modified silica nanoparticles (NPs) for the delivery of the second-generation photosensitizer meta-tetra(hydroxyphenyl)chlorin (mTHPC) to cancer cells in vitro. mTHPC was entrapped in NPs (~33 nm diameter) in a monomeric form which produced singlet oxygen with a high efficiency. In aqueous media with high salt concentrations, the NPs underwent aggregation and precipitation but their stability could be preserved in the presence of foetal bovine serum. The cellular uptake, localization and phototoxic activity of mTHPC was determined comparatively in human oesophageal cancer cells after its delivery by the NPs and the standard solvent ethanol/poly(ethylene glycol) 400/water (20:30:50, by vol). The NP formulation reduced the cellular uptake of mTHPC by about 50% in comparison to standard solvent while it did not affect the concentration-dependent photokilling activity of mTHPC and its intracellular localization. Fluorescence resonance energy transfer measurements, using NPs with mTHPC physically entrapped and a cyanine covalently linked, and ultracentrifugation experiments indicated that mTHPC is transferred from NPs to serum proteins when present in the medium. However, the coating of the NP surface with poly(ethylene glycol) largely prevented the transfer to proteins. In conclusion, mTHPC is rapidly transferred from the uncoated nanoparticles to the serum proteins and then internalized by the cells as a protein complex, irrespective of its modality of delivery.

Compagnin, Chiara; Baù, Luca; Mognato, Maddalena; Celotti, Lucia; Miotto, Giovanni; Arduini, Maria; Moret, Francesca; Fede, Caterina; Selvestrel, Francesco; Rio Echevarria, Iria M.; Mancin, Fabrizio; Reddi, Elena

2009-08-01

327

Free serum concentrations of the protein-bound retention solute p-cresol predict mortality in hemodialysis patients.  

PubMed

Based on in vitro data, protein-bound uremic retention solutes have increasingly been recognized to play a pathophysiological role in the uremic syndrome. p-Cresol, a representative of this group of molecules, has been shown to be implicated in uremic immunodeficiency and endothelial dysfunction, potentially linking its serum levels to mortality. Thus far, however, no clinical information on this issue is available. To determine the relationship between p-cresol and all-cause mortality, 175 prevalent hemodialysis (HD) patients were enrolled in a prospective study. At baseline, serum levels of the water-soluble solutes urea, creatinine, and phosphate, the middle molecule beta2-microglobulin, total and free concentrations of the protein-bound solute p-cresol, and several risk factors for mortality were evaluated. During a median follow-up of 34 months, 60 patients died. Baseline comorbidity (Davies score) (hazard ratio (HR), 1.49; 95% confidence interval (95% CI), 1.19-1.86), impaired nutritional status (HR, 4.22; 95% CI, 2.15-8.29), time since initiation of dialysis (HR, 0.98; 95% CI, 0.97-1.00), and higher free concentrations of the protein-bound solute p-cresol (HR, 2.28; 95% CI, 1.12-4.64) were independently associated with mortality (multivariate Cox proportional hazards analysis). Our data suggest that free serum levels of p-cresol, a representative of the protein-bound uremic retention solutes, are associated with mortality in HD patients. These findings may encourage nephrologists to widen their field of interest beyond the scope of small water-soluble uremic solutes and middle molecules. PMID:16421516

Bammens, B; Evenepoel, P; Keuleers, H; Verbeke, K; Vanrenterghem, Y

2006-03-01

328

Monocyte chemoattractant protein-1 gene polymorphism and its serum level have an impact on anthropometric and biochemical risk factors of metabolic syndrome in Indian population.  

PubMed

Monocyte chemoattractant protein-1 (MCP-1), encoded by gene CCL-2 (Chemokine C-C motif 2), is the ligand of chemokine receptor CCR-2. Concurrent clinical alteration in several metabolic aspects, including central obesity, dysglycemia, dyslipidemia and hypertension, is clinically characterized as metabolic syndrome (MetS). Role of MCP-1 in each of these aspects has been established in vitro and in animal studies as well. We here report genetic association of -2518 A>G MCP-1 (rs 1024611) gene polymorphism and level of MCP-1 with MetS in North Indian subjects. We analysed (n = 386, controls and n = 384, MetS subjects) for MCP-1 gene polymorphism using PCR-RFLP, its serum level using ELISA, anthropometric (body mass index, waist and hip circumferences, waist-hip ratio and blood pressure) and biochemical (serum lipids, plasma glucose and insulin levels) variables in a genetic association study. The body mass index, waist circumference, hip circumference, waist-hip ratio, blood pressure, serum lipids, insulin and fasting plasma glucose level were significantly high in MetS subjects. Regression analysis showed significant correlation of body mass index, waist and hip circumference, systolic/diastolic blood pressure, fasting glucose, total cholesterol, high-density lipoprotein, low-density lipoprotein fasting insulin and HOMA-IR with MetS. MCP-1 allele and genotype were significantly associated with MetS. Serum MCP-1 level was high in overall cases. In conclusions, the MCP-1 2518A>G (rs 1024611) polymorphism has significant impact on risk of MetS, and MCP-1 level correlates with anthropometric and biochemical risk factors of MetS. PMID:25639755

Madeshiya, A K; Singh, S; Dwivedi, S; Saini, K S; Singh, R; Tiwari, S; Konwar, R; Ghatak, A

2015-04-01

329

Prognostic significance of low serum levels of Clara cell phospholipid-binding protein in occupational aluminium neurotoxicity.  

PubMed

The relationship between respiratory and neurological effects of exposure to aluminium (Al) was investigated in a group of foundry workers exposed to Al at concentrations below the threshold limit value (TLV) binding in Poland (2.0 mg Al2O3 m(-3)). Neurological and neurophysiological parameters indicated subclinical effects of Al exposure on the nervous system. The measurement of serum anti-inflammatory Clara cell protein (CC16) was employed as a peripheral marker of the lung epithelium function. There was a strong inverse relationship between serum Al (Al-S) and CC16 concentrations (p = 0.006). The lowest CC16 concentrations were found in serum of workers characterised by subjective symptoms of the central nervous system (CNS) and abnormal results of neurophysiological examinations (EEG and VEP). Low serum CC16 concentrations and enhanced Al and iron (Fe) levels were also observed in the younger age group of workers with the subjective CNS symptoms and abnormal VEP results, which suggests that Fe is implicated in strengthening of the neurotoxic Al potential. The results of our study support the hypothesis that subclinical neurological symptoms (especially abnormal VEP) are most likely associated with internalisation of Al ions with lipid fractions of the lung epithelium, which in turn may help Al ions overcome the blood-brain barrier. Low serum CC16 concentrations (<10 microg L(-1)) were noted in workers with the abnormal results of neurological (CNS) and neurophysiological (EEG and VEP) examinations as well as with Al body burden manifested by urinary excretion (Al-U) below 60 microg L(-1) and Al-S concentration of 2 microg L(-1). This concentration may be considered as a threshold allowable biological concentration of aluminium. PMID:16099050

Halatek, Tadeusz; Sinczuk-Walczak, H; Rydzynski, K

2005-09-01

330

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

331

Calcium-dependent and -independent binding of the pentraxin serum amyloid P component to glycosaminoglycans and amyloid proteins: enhanced binding at slightly acid pH.  

PubMed

Serum amyloid P component (SAP), a member of the pentraxin family of proteins, binds calcium-dependently to several ligands including glycosaminoglycans (GAG's). We have investigated the influence of pH on the Ca2(+)-dependent binding of SAP to solid phase GAG's and amyloid fibril proteins (AA and beta2M) by ELISA. An increase in the dose-dependent binding of SAP to heparan sulfate, AA-protein and beta2M was observed as the pH decreased from 8.0 to 5.0. Furthermore, a lower, but significant Ca2(+)-independent binding of SAP to heparan sulfate, dermatan sulfate, AA protein and the amyloid precursor protein beta2M was observed. This binding was also enhanced at slightly acid pH, most pronounced at pH 5.0. The results of this study indicate that SAP can exhibit both Ca2(+)-dependent and -independent binding to ligands involved in amyloid fibril formation and that the binding is enhanced under conditions of slightly lowered pH. PMID:9165101

Danielsen, B; Sørensen, I J; Nybo, M; Nielsen, E H; Kaplan, B; Svehag, S E

1997-04-25

332

Rho GTPase protein expression and activation in murine monocytes/macrophages is not modulated by model biomaterial surfaces in serum-containing in vitro cultures  

PubMed Central

The Rho GTPase cellular signaling cascade was investigated in pro-monocyte and (monocyte-)macrophage cells by examining GTPase expression and activation in serum-containing cultures on model biomaterials. Abundance of Rho GDI and the Rho GTPase proteins RhoA, Cdc42 and Rac1 was determined in cells grown on tissue culture polystyrene, polystyrene, poly-l-lactide and Teflon® AF surfaces. Protein expression was compared based on cell maturity (pro-monocyte to monocyte to macrophage lineages) and by model surface chemistry: Rho proteins were present in the majority of macrophage cells tested on model surfaces suggesting that a pool of Rho proteins is readily available for signaling events in response to numerous activating cues, including biomaterials surface encounter. Rho GTPase activation profiles in these cell lines indicate active Cdc42 and Rho proteins in RAW 264.7, Rac1 and Rho in J774A.1, and Cdc42 and Rac1 in IC-21 cell lines, respectively. Collectively, these proteins are known to play critical roles in all actin-based cytoskeletal rearrangement necessary for cell adhesion, spreading and motility, and remain important to establishing cellular responses required for foreign body reactions in vivo. Differences in Rho GTPase protein expression levels based on cell sourcing (primary versus secondary-derived cell source), or as a function of surface chemistry were insignificant. Rho GTPase expression profiles varied between pro-monocytic non-adherent precursor cells and mature adherent monocyte/macrophage cells. The active GTP-bound forms of the Rho GTPase proteins were detected from monocyte-macrophage cell lines RAW 264.7 and J774A.1 on all polymer surfaces, suggesting that while these proteins are central to cell adhesive behavior, differences in surface chemistry are insufficient to differentially regulate GTPase activation in these cell types. Active Cdc42 was detected from cells cultured on the more-polar tissue culture polystyrene and poly-l-lactide surfaces after several days, but absent from those grown on apolar polystyrene and Teflon® AF, indicating some surface influence on this GTPase in serum-containing cultures. PMID:17235380

GODEK, M. L.; SAMPSON, J. A.; DUCHSHERER, N. L.; McELWEE, Q.; GRAINGER, D. W.

2006-01-01

333

Influence of renal impairment, chemical form, and serum protein binding on intravenous and oral aluminum kinetics in the rabbit.  

PubMed

The influence of renal impairment on the intravenous kinetics of aluminum (Al) lactate and the oral absorption of eight representative Al forms was determined. The serum protein binding of Al was assessed. Creatinine clearance in renally impaired rabbits was 23% of controls. Systemic clearance of Al was less in renally impaired rabbits (39 vs. 53 ml/hr/kg), as were the steady-state volume of distribution (516 vs. 1175 ml/kg), the half-life of elimination (14 vs. 27 hr), and the mean residence time of Al (14 vs. 25 hr). The shorter Al half-life and mean residence time in renally impaired rabbits were due to a diminished volume of Al distribution. Oral bioavailability of Al in renally intact rabbits ranged from 0.3 to 2.2% (Al borate less than glycinate less than hydroxide less than chloride less than sucralfate less than lactate less than nitrate less than citrate). Renal impairment had little influence on oral bioavailability of most Al forms, although it increased Al citrate absorption to 4.6%. In vitro and in vivo determination of Al ultrafilterability (less than 30,000 D) as an estimate of serum protein binding suggested a greater percentage of ultrafilterable Al species in renally impaired rabbit serum than in control rabbit serum. The increase in ultrafilterable Al species produced the less than expected reduction in Al clearance in renally impaired rabbits. The ultrafilterability of various Al concentrations was greater for citrate greater than lactate greater than nitrate greater than chloride, perhaps partially explaining the similar rank order of oral absorption of these Al forms. The physicochemistry of the eight Al forms was further characterized by determination of their octanol/water partitioning coefficients and their water solubility. There was a significant correlation between the percentage absorbed and the log of the octanol/water partition coefficient. Knowledge of the physicochemistry of Al aids in the understanding of Al pharmacokinetics. PMID:3413794

Yokel, R A; McNamara, P J

1988-08-01

334

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

335

DECREASED HEART RATE IS ASSOCIATED WITH CARBAMATE-INDUCED ACTIVATION OF PRO-INFLAMMATORY SERUM PROTEINS.  

EPA Science Inventory

Previously we reported that chlorpyrifos (CHP), an irreversible cholinesterase (ChE) inhibitor, induces hypertension in rats. Concomitant with hypertension, we found an increase in C-reactive protein, macrophage inflammatory protein-2 , monocyte chemotactic protein-5 and interfer...

336

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

337

Serum-free production of recombinant proteins and adenoviral vectors by 293SF-3F6 cells.  

PubMed

This article describes the step-wise approach undertaken to select a serum-free medium (SFM) for the efficient production of a recombinant adenoviral vectors expressing beta-galactosidase (Ad5 CMV-LacZ), in the complementing human embryonic kidney 293S cells. In the first step, a 293S-derived transfectoma, secreting a soluble epidermal growth factor receptor sEGFr (D2-22), was used to estimate the potential of selected serum-free formulations to support the production of a recombinant protein as compared to serum-containing medium. Assays showed that only one among six commercial serum-free formulations could support both sEGFr production and cell growth in static or suspension culture. In commercially available calcium-containing serum-free formulations, the cell aggregates reached up to 3 mm in diameter. In the second step, 293S cells were gradually adapted to a low-calcium version of the selected medium (LC-SFM). Cells were cloned, then screened according to their ability to grow at a rate and an extent comparable to parental cells in serum-containing medium (standard) as single cells or small aggregates. The 293SF-3F6 clone, first adapted to and then cloned in the selected serum-free medium, was selected for further experiments. Bioreactor run performed with the 293SF-3F6 clone showed similar growth curve as in the shake-flask controls. In the final step, the recombinant viral vector productivity of the 293S cells and the 293SF-3F6 clone was tested. The 293SF-3F6 cells infected by Ad5 CMV-LacZ in 3 L-scale bioreactor maintained the specific productivities of both beta-galactosidase and adenoviral vector equivalent to the shake-flask controls in suspension culture. Results from this study clearly demonstrate that the 293SF-3F6 cell line thus selected may be used either for establishing stable transfected cell line or for the production of adenoviral vectors required for gene therapy studies. PMID:10099373

Côté, J; Garnier, A; Massie, B; Kamen, A

1998-09-01

338

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

339

Gene-specific DNA methylation association with serum levels of C-reactive protein in African Americans.  

PubMed

A more thorough understanding of the differences in DNA methylation (DNAm) profiles in populations may hold promise for identifying molecular mechanisms through which genetic and environmental factors jointly contribute to human diseases. Inflammation is a key molecular mechanism underlying several chronic diseases including cardiovascular disease, and it affects DNAm profile on both global and locus-specific levels. To understand the impact of inflammation on the DNAm of the human genome, we investigated DNAm profiles of peripheral blood leukocytes from 966 African American participants in the Genetic Epidemiology Network of Arteriopathy (GENOA) study. By testing the association of DNAm sites on CpG islands of over 14,000 genes with C-reactive protein (CRP), an inflammatory biomarker of cardiovascular disease, we identified 257 DNAm sites in 240 genes significantly associated with serum levels of CRP adjusted for age, sex, body mass index and smoking status, and corrected for multiple testing. Of the significantly associated DNAm sites, 80.5% were hypomethylated with higher CRP levels. The most significant Gene Ontology terms enriched in the genes associated with the CRP levels were immune system process, immune response, defense response, response to stimulus, and response to stress, which are all linked to the functions of leukocytes. While the CRP-associated DNAm may be cell-type specific, understanding the DNAm association with CRP in peripheral blood leukocytes of multi-ethnic populations can assist in unveiling the molecular mechanism of how the process of inflammation affects the risks of developing common disease through epigenetic modifications. PMID:23977389

Sun, Yan V; Lazarus, Alicia; Smith, Jennifer A; Chuang, Yu-Hsuan; Zhao, Wei; Turner, Stephen T; Kardia, Sharon L R

2013-01-01

340

Cryopreservation for bovine embryos in serum-free freezing medium containing silk protein sericin.  

PubMed

Because the use of serum in the embryo cryopreservation increases the probability of animal health problems such as bovine spongiform encephalopathy (BSE) and viral infections, this study was conducted to examine the effects of sericin supplementation for serum-free freezing medium on the survival and development of bovine embryos after freezing-thawing and direct transfer to recipients. When in vitro-produced bovine embryos were frozen conventionally in the freezing medium supplemented with various concentrations (0.1%, 0.5%, and 1.0%) of sericin, the percentages of damaged zona pellucida, survival, and development of frozen-thawed embryos were similar to those of embryos frozen in freezing medium supplemented with 0.4% bovine serum albumin (BSA) and 20% fetal bovine serum (FBS) (0.4BSA/20F; control). When in vivo-derived embryos were frozen with 0.4BSA/20F (control), 0.5% sericin +20% FBS (0.5S/20F) or 0.5% sericin (0.5S) and were subsequently transferred directly to recipients, the percentages of recipients with pregnancy and normal calving in the 0.5S/20F group were higher than those in the control group (47.3% vs. 40.1% and 94.6% vs. 87.3%, respectively). Moreover, the percentages of recipients with pregnancy and normal calving (42.2% and 92.4%, respectively) in the 0.5S group were similar with those of other groups. In conclusion, these results indicate that serum-free freezing medium supplemented with sericin is available for the cryopreservation of bovine embryos and that it is beneficial for the elimination of a potential source of biological contamination by serum or BSA. PMID:23850826

Isobe, Tomohiro; Ikebata, Yoshihisa; Onitsuka, Takeshi; Do, Lanh Thi Kim; Sato, Yoko; Taniguchi, Masayasu; Otoi, Takeshige

2013-10-01

341

Serum protein layers on parylene-C and silicon oxide: effect on cell adhesion  

E-print Network

for adsorption of Fn at 50 #2;g/mL (A) and alb actor) supplemented DMEM (Dulbecco’s modified Eagle edia (10% fetal calf serum, 1% penicillin/streptomycin, ine, 100 #2;M 2-mercaptoethanol). The mES cells were d split (ratio 1:8) every 2 days. On day 0 mES cells... were non-tissue culture treated petri-dishes at a density of /mL and allowed to aggregate into embryoid bodies (EB) DMEM media (ADMEM/F12:neurobasal medium (1:1), ut serum replacement, 1% penicillin/streptomycin, 1% , 100 #2;M 2-mercaptoethanol...

Delivopoulos, Evangelos; Ouberai, Myriam M.; Coffey, Paul D.; Swann, Marcus J.; Shakesheff, Kevin M.; Welland, Mark E.

2014-12-16

342

Acute Sleep Deprivation Increases Serum Levels of Neuron-Specific Enolase (NSE) and S100 Calcium Binding Protein B (S-100B) in Healthy Young Men  

PubMed Central

Study Objectives: To investigate whether total sleep deprivation (TSD) affects circulating concentrations of neuron-specific enolase (NSE) and S100 calcium binding protein B (S-100B) in humans. These factors are usually found in the cytoplasm of neurons and glia cells. Increasing concentrations of these factors in blood may be therefore indicative for either neuronal damage, impaired blood brain barrier function, or both. In addition, amyloid ? (A?) peptides 1-42 and 1-40 were measured in plasma to calculate their ratio. A reduced plasma ratio of A? peptides 1-42 to 1-40 is considered an indirect measure of increased deposition of A? 1-42 peptide in the brain. Design: Subjects participated in two conditions (including either 8-h of nocturnal sleep [22:30-06:30] or TSD). Fasting blood samples were drawn before and after sleep interventions (19:30 and 07:30, respectively). Setting: Sleep laboratory. Participants: 15 healthy young men. Results: TSD increased morning serum levels of NSE (P = 0.002) and S-100B (P = 0.02) by approximately 20%, compared with values obtained after a night of sleep. In contrast, the ratio of A? peptides 1-42 to 1-40 did not differ between the sleep interventions. Conclusions: Future studies in which both serum and cerebrospinal fluid are sampled after sleep loss should elucidate whether the increase in serum neuron-specific enolase and S100 calcium binding protein B is primarily caused by neuronal damage, impaired blood brain barrier function, or is just a consequence of increased gene expression in non-neuronal cells, such as leukocytes. Citation: Benedict C; Cedernaes J; Giedraitis V; Nilsson EK; Hogenkamp PS; Vågesjö E; Massena S; Pettersson U; Christoffersson G; Phillipson M; Broman JE; Lannfelt L; Zetterberg H; Schiöth HB. Acute Sleep Deprivation Increases Serum Levels of Neuron-Specific Enolase (NSE) and S100 Calcium Binding Protein B (S-100B) in Healthy Young Men. SLEEP 2014;37(1):195-198. PMID:24470708

Benedict, Christian; Cedernaes, Jonathan; Giedraitis, Vilmantas; Nilsson, Emil K.; Hogenkamp, Pleunie S.; Vågesjö, Evelina; Massena, Sara; Pettersson, Ulrika; Christoffersson, Gustaf; Phillipson, Mia; Broman, Jan-Erik; Lannfelt, Lars; Zetterberg, Henrik; Schiöth, Helgi B.

2014-01-01

343

Ligusticum chuanxiong prevents rat pheochromocytoma cells from serum deprivation-induced apoptosis through a protein kinase A-dependent pathway.  

PubMed

Ligusticum chuanxiong (LC) is a traditional Chinese herbal medicine used to treat various cardiovascular diseases. In this study, the butanol extract of LC was found to protect neuronal-like pheochromocytoma cells from serum deprivation-induced apoptosis. Both a serine/threonine kinase inhibitor and a specific protein kinase A (PKA) inhibitor blocked the protective effect of LC. A transcription inhibitor (actinomycin D) and a protein synthesis inhibitor (cyclohexamide) also attenuated the protective effect of LC, suggesting the requirement of gene expression for the protection of LC. On the other hand, LC increased both the formation of cyclic-AMP and the phosphorylation of the cyclic-AMP response element-binding protein (CREB), a downstream target of PKA and a nuclear transcription factor known for neuroprotective mechanism. Furthermore, LC-induced CREB phosphorylation and protective effect could be blocked by a PKA inhibitor and overexpression of the dominant negative CREB, respectively. Taken together, the protective mechanism of LC in antagonizing serum deprivation-induced PC12 cell apoptosis might be mediated through a PKA/CREB-dependent pathway. PMID:16973320

Lin, Yun-Lian; Lee, Yi-Chao; Huang, Chuen-Lin; Lai, Wen-Lin; Lin, Yen-Ru; Huang, Nai-Kuei

2007-02-12

344

Chemistry-dependent adsorption of serum proteins onto polyanhydride microparticles differentially influences dendritic cell uptake and activation.  

PubMed

The delivery of antigen-loaded microparticles to dendritic cells (DCs) may benefit from surface optimization of the microparticles themselves, thereby exploiting the material properties and introducing signals that mimic pathogens. Following in vivo administration microparticle surface characteristics are likely to be significantly modified as proteins are quickly adsorbed onto their surface. In this work we describe the chemistry-dependent serum protein adsorption patterns on polyanhydride particles and the implications for their molecular interactions with DCs. The enhanced expression of MHC II and CD40 on DCs after incubation with amphiphilic polyanhydride particles, and the increased secretion of IL-6, TNF-?, and IL-12p40 by hydrophobic polyanhydride particles exemplified the chemistry-dependent activation of DCs by sham-coated particles. The presence of proteins such as complement component 3 and IgG further enhanced the adjuvant properties of these vaccine carriers by inducing DC maturation (i.e. increased cell surface molecule expression and cytokine secretion) in a chemistry-dependent manner. Utilizing DCs derived from complement receptor 3-deficient mice (CR3(-/-) mice) identified a requirement for CR3 in the internalization of both sham- and serum-coated particles. These studies provide valuable insights into the rational design of targeted vaccine platforms aimed at inducing robust immune responses and improving vaccine efficacy. PMID:22684115

Carrillo-Conde, Brenda R; Ramer-Tait, Amanda E; Wannemuehler, Michael J; Narasimhan, Balaji

2012-10-01

345

Each row in the table corresponds to one protein group. A. Protein IDs: SGD ORFs, including all proteins that could be identified from the peptides identified.  

E-print Network

: SGD ORFs, including all proteins that could be identified from the peptides number of peptide identified. C. SGD Id: Mapping from first id to SGD identifier D. Gene Name: Mapped from SGD E. ControlCDC48 Ratio: Ratio of cdc48

Martin, Alain

346

Investigation of serum protein profiles in scrapie infected sheep by means of SELDI-TOF-MS and multivariate data analysis  

PubMed Central

Background Classical scrapie in sheep is a fatal neurodegenerative disease associated with the conversion PrPC to PrPSc. Much is known about genetic susceptibility, uptake and dissemination of PrPSc in the body, but many aspects of prion diseases are still unknown. Different proteomic techniques have been used during the last decade to investigate differences in protein profiles between affected animals and healthy controls. We have investigated the protein profiles in serum of sheep with scrapie and healthy controls by SELDI-TOF-MS and LC-MS/MS. Latent Variable methods such as Principal Component Analysis, Partial Least Squares-Discriminant Analysis and Target Projection methods were used to describe the MS data. Results The serum proteomic profiles showed variable differences between the groups both throughout the incubation period and at the clinical end stage of scrapie. At the end stage, the target projection model separated the two groups with a sensitivity of 97.8%, and serum amyloid A was identified as one of the protein peaks that differed significantly between the groups. Conclusions At the clinical end stage of classical scrapie, ten SELDI peaks significantly discriminated the scrapie group from the healthy controls. During the non-clinical incubation period, individual SELDI peaks were differently expressed between the groups at different time points. Investigations of differences in -omic profiles can contribute to new insights into the underlying disease processes and pathways, and advance our understanding of prion diseases, but comparison and validation across laboratories is difficult and challenging. PMID:24229425

2013-01-01

347

Systemic and lung protein changes in sarcoidosis. Lymphocyte counts, gallium uptake values, and serum angiotensin-converting enzyme levels may reflect different aspects of disease activity  

SciTech Connect

BAL lymphocyte percentages, quantitated gallium-67 lung uptake, and SACE levels have all been proposed as measures of disease activity in sarcoidosis. We analyzed 32 paired sera and BAL fluids from sarcoidosis patients by high-resolution agarose electrophoresis to look for protein changes characteristic of systemic or local inflammation and compared the results with those from the above tests. Nine patients (group 1) had serum inflammatory protein changes and increased total protein, albumin, beta 1-globulin (transferrin), and gamma-globulin levels in fluid recovered by BAL. Thirteen patients (group 2) had normal protein levels in sera but abnormal protein levels in BAL specimens. Ten patients (group 3) had normal protein levels in sera and in BAL specimens. Patients in groups 1 and 2 had a disproportionate increase in beta 1-globulin (transferrin) and gamma-globulin levels in their BAL specimens. The BAL lymphocyte percentage changes paralleled the BAL protein level changes, suggesting relationships among the immunoregulatory role of these cells, increased local immunoglobulin synthesis, and the pathogenesis of altered alveolar permeability. Gallium-67 uptake was highest in patients with serum inflammatory protein changes. Thus, systemic inflammation may facilitate pulmonary gallium-67 uptake, possibly by changes in BAL fluid or serum transferrin saturation and/or kinetics. SACE levels showed no relationship to changes in the levels of serum or BAL proteins. These data suggest that the various proposed measures of disease activity reflect different aspects of inflammation in sarcoidosis.

Check, I.J.; Kidd, M.R.; Staton, G.W. Jr.

1986-01-01

348

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

349

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

350

Preparation and characterization of a novel exendin-4 human serum albumin fusion protein expressed in Pichia pastoris.  

PubMed

A novel recombinant exendin-4 human serum albumin fusion protein (rEx-4/HSA) expressed in Pichia pastoris was prepared and characterized. Ex-4 is a 39-amino acid peptide isolated from the salivary gland of the lizard Heloderma suspectum and is thought to be a novel therapeutic agent for type 2 diabetes. But to gain a continued effect, the peptide has to be injected twice a day owing to its short plasma half-life (t(1/2) = 2.4 h). To extend the half-life of Ex-4 molecule in vivo, we designed a genetically engineered Ex-4/HSA fusion protein. Between Ex-4 and HSA, a peptide linker GGGGS was inserted and the fusion protein was expressed in methylotrophic yeast P. pastoris with native HSA secretion signal sequence. The recombinant protein was secreted correctly and was obtained with high purity (typically > 98%) by a three-step purification procedure. cAMP assay demonstrated that the fusion protein had a bioactivity similar to Ex-4 for interaction with GLP-1 receptors in vitro. Results from oral glucose tolerance test indicated that rEx-4/HSA could effectively improve glucose tolerance in diabetic db/db mice. Pharmacokinetics studies in cynomologus monkeys also showed that rEx-4/HSA had a much longer plasma half-life. Therefore, rEx-4/HSA fusion protein could potentially be used as a new recombinant biodrug for type 2 diabetes therapy. PMID:17994612

Huang, Yan-Shan; Chen, Zhi; Chen, Yi-Qiong; Ma, Guo-Chang; Shan, Jian-Feng; Liu, Wei; Zhou, Lin-Fu

2008-05-01

351

Serum Proteins and Alkaline Phosphatase Levels in Patients with Tuberous Sclerosis  

ERIC Educational Resources Information Center

Six 4- to 37-year-old patients with tuberosis sclerosis (a chronic condition characterized by siezures, intercranial calcification, a reddish-yellow sebaceous glandular mass on the face, and frequent crises in early years), did not exhibit an elevation of the (alpha + beta) globulin fraction in their serum. (Author/MC)

Fischer, M. H.; And Others

1974-01-01

352

Polymorphism of serum proteins (C3, BF, HP and TF) of six populations in Colombia.  

PubMed

Five hundred and eighty-five serum samples from six populations in Colombia (Baranoa, Choco, Uitoto Indians, Subachoque, Pasto and Urban Bogotan) were investigated for four genetic markers. For the HP, C3 and BF systems but not for TF there is a wide range of gene frequency variation and these differences are compared with those in the few previous studies. PMID:2759637

Bernal, J E; Sarmiento, P; Briceno, I; Papiha, S S

1989-01-01

353

Regulation of heat shock protein message in Jurkat cells cultured under serum-starved and gravity-altered conditions  

NASA Technical Reports Server (NTRS)

Although our understanding of effects of space flight on human physiology has advanced significantly over the past four decades, the potential contribution of stress at the cellular and gene regulation level is not characterized. The objective of this ground-based study was to evaluate stress gene regulation in cells exposed to altered gravity and environmentally suboptimal conditions. We designed primers to detect message for both the constitutive and inducible forms of the heat shock protein, HSP-70. Applying the reverse transcriptase-polymerase chain reaction (RT-PCR), we probed for HSP-70 message in human acute T-cell leukemia cells, Jurkat, subjected to three types of environmental stressors: (1) altered gravity achieved by centrifugation (hypergravity) and randomization of the gravity vector in rotating bioreactors, (2) serum starvation by culture in medium containing 0.05% serum, and (3) temperature elevation (42 degrees C). Temperature elevation, as the positive control, significantly increased HSP-70 message, while centrifugation and culture in rotating bioreactors did not upregulate heat shock gene expression. We found a fourfold increase in heat shock message in serum-starved cells. Message for the housekeeping genes, actin and cyclophilin, were constant and comparable to unstressed controls for all treatments. We conclude that gravitational perturbations incurred by centrifugal forces, exceeding those characteristic of a Space Shuttle launch (3g), and culture in rotating bioreactors do not upregulate HSP-70 gene expression. In addition, we found RT-PCR useful for evaluating stress in cultured cells. Copyright 2000 Wiley-Liss, Inc.

Lewis, M. L.; Hughes-Fulford, M.

2000-01-01

354

Serum thymidine kinase 1 and C-reactive protein as biomarkers for screening clinically healthy dogs for occult disease.  

PubMed

Thymidine kinase (TK1) is a biomarker that correlates well with diagnosis and prognosis in certain canine cancers. Canine C-reactive protein (cCRP) is a widely accepted marker of inflammation correlated with increased risk and severity of various diseases. We evaluated serum TK1 and cCRP concentrations in apparently healthy dogs (n?=?360). All dogs were followed up for a minimum of 6?months by health questionnaire. All dogs with cancer were identified using a proprietary dual-biomarker algorithm [termed Neoplasia Index (NI)]. Specificity of positive NI is 0.91 and high positive is 0.98. All-cause mortality was 20% in dogs with elevated cCRP and 3% in dogs with low cCRP. The performance of serum TK1 and cCRP as tools for screening for occult cancer is improved when evaluated together. Serum TK1 and cCRP (unified in the NI) are useful in the screening of occult canine cancer. cCRP is useful in screening for other serious diseases. PMID:23859156

Selting, K A; Sharp, C R; Ringold, R; Knouse, J

2013-07-16

355

DNA-protein interactions between mammalian nuclear proteins and a GCC-element included in a composite cis-acting element of mouse ribosomal protein L32 promoter.  

PubMed

DNA-protein complex formation between the sequence GC(GCC)4 (GCC-element) of mouse ribosomal protein L32 (rpL32) promoter and nuclear proteins of mouse and human cells has been studied using gel retardation and South-Western blotting methods. The rpL32 promoter fragment (-24.+11) was able to form specific complexes with mouse and human nuclear proteins mainly due to the presence of the GCC-element (-19.-6). DNA-protein complex patterns exhibited marked tissue-specificity. Three nuclear polypeptides of approximately 18, 28, and 50 kD that bind to the rpL32 promoter region (-24.+11) have been detected in HeLa cells by ligand blotting. At least one of them (18 kD) interacted with the GCC-element directly. The same fragment of the promoter interacted only with one nuclear polypeptide (28-31 kD) from human fibroblasts. DNA-protein complex formation between the investigated rpL32 promoter fragment containing the GCC-element and human fibroblast nuclear proteins is Zn2+-dependent. The method of functional titration (in vivo competition in the CAT-test) revealed that the GCC-element within the rpL32 promoter functions as a positive cis-acting transcriptional element in NIH 3T3 cells. Thus, our data characterize the sequence GC(GCC)4 as a functionally active cis-element included as a component in the more complex (composite) cis-element of mouse rpL32 promoter exhibiting tissue-specific properties. In various mammalian cell types the GCC-element can interact with various nuclear proteins, and the mode of these interactions can be determined by its relative position to other cis-elements in the regulatory sites of the genome. PMID:10187914

Orlov, S V; Kuteikin, K B; Dizhe, E B; Kuryshev, V Y; Shpakovich, V M; Perevozchikov, A P

1999-02-01

356

Effects of serum amyloid A protein (SAA) on composition, size, and density of high density lipoproteins in subjects with myocardial infarction  

Microsoft Academic Search

The acute phase reactant serum amyloid A protein (SAA) circulates in plasma as a constituent of high density lipo- proteins (HDL). Advantage has been taken of the induction of SAA in human subjects with myocardial infarction to study the effect of SAA on the physical and chemical properties of HDL. HDL were isolated by sequential ultracentrifugation and assayed for chemical

Peter M. Clifton; A. Malcolm Mackinnon; Phillip J. Barter

357

Role of serum and induced sputum surfactant protein D in predicting the response to treatment in chronic obstructive pulmonary disease  

PubMed Central

This study was designed to determine the expression of serum and sputum surfactant protein D (SP-D) in chronic obstructive pulmonary disease (COPD) and its association with treatment response. Sixty-five treatment-naive patients with COPD and 26 normal control subjects were recruited in the study. The concentrations of serum and sputum SP-D were measured, and the associations of SP-D with pulmonary function and the modified Medical Research Council dyspnea scale (mMRC) and the St. George’s Respiratory Questionnaire (SGRQ) scores before and after three months of treatment with an inhaled corticosteroid and a long-acting ?2-agonist were analyzed. The concentrations of serum and sputum SP-D in the COPD group (45.46±37.78 and 173.23±186.93 ng/ml, respectively) were significantly higher than those of the normal control group (31.68±12.04 and 89.59±70.29 ng/ml, respectively). After three months of treatment, serum SP-D levels were reduced to 30.7±13.9 ng/ml and were significantly lower than the baseline levels (t=2.217, P=0.031). However, no significant reduction in sputum SP-D levels was observed following the treatment (P>0.05). A significant association between baseline sputum SP-D and change in SGRQ activity scores (r=?0.652, P=0.012) was observed; however no association was established with the changes in other clinical profiles following the treatment (P>0.05). This result suggested that an increased baseline sputum SP-D may be a weak predictive indicator of response to treatment with inhaled corticosteroids and long-acting ?2-agonists in patients with COPD. PMID:25187846

LIU, WEI; JU, CHUN-RONG; CHEN, RONG-CHANG; LIU, ZHI-GUANG

2014-01-01

358

Cultivation of Bowes melanoma cells in the Opticell system: influence of the addition of protein supplements to the serum-free medium on the production of plasminogen activator.  

PubMed

The influence of a protein-free and a protein-rich, supplemented serum-free medium on the production of plasminogen activator (t-PA) from Bowes melanoma cells was investigated in the Opticell culture system and compared to tissue culture flask cultures. In the presence of medium supplements metabolic activity and t-PA production were favoured in both systems. The addition of supplements was apparently more effective in the Opticell than in flask cultures, because t-PA activity obtained in the Opticell was 2-3 times higher in protein-rich medium, but 2 times lower in unsupplemented medium than in flasks. These results indicate that the protein content in a serum-free medium is important for product formation in the Opticell, and serum-free media which work at small scale in tissue culture flasks are not always suited for technical culture systems such as the Opticell but have to be adapted to them. PMID:3108052

Bödeker, B G; Klimetzek, V; Klein, U; Hewlett, G; Schlumberger, H D

1987-01-01

359

N-Glycomic Changes in Serum Proteins in Type 2 Diabetes Mellitus Correlate with Complications and with Metabolic Syndrome Parameters  

PubMed Central

Background Glycosylation, i.e the enzymatic addition of oligosaccharides (or glycans) to proteins and lipids, known as glycosylation, is one of the most common co-/posttranslational modifications of proteins. Many important biological roles of glycoproteins are modulated by N-linked oligosaccharides. As glucose levels can affect the pathways leading to glycosylation of proteins, we investigated whether metabolic syndrome (MS) and type 2 diabetes mellitus (T2DM), pathological conditions characterized by altered glucose levels, are associated with specific modifications in serum N-glycome. Methods We enrolled in the study 562 patients with Type 2 Diabetes Mellitus (T2DM) (mean age 65.6±8.2 years) and 599 healthy control subjects (CTRs) (mean age, 58.5±12.4 years). N-glycome was evaluated in serum glycoproteins. Results We found significant changes in N-glycan composition in the sera of T2DM patients. In particular, ?(1,6)-linked arm monogalactosylated, core-fucosylated diantennary N-glycans (NG1(6)A2F) were significantly reduced in T2DM compared with CTR subjects. Importantly, they were equally reduced in diabetic patients with and without complications (P<0.001) compared with CTRs. Macro vascular-complications were found to be related with decreased levels of NG1(6)A2F. In addition, NG1(6)A2F and NG1(3)A2F, identifying, respectively, monogalactosylated N-glycans with ?(1,6)- and ?(1,3)-antennary galactosylation, resulted strongly correlated with most MS parameters. The plasmatic levels of these two glycans were lower in T2DM as compared to healthy controls, and even lower in patients with complications and MS, that is the extreme “unhealthy” phenotype (T2DM+ with MS). Conclusions Imbalance of glycosyltransferases, glycosidases and sugar nucleotide donor levels is able to cause the structural changes evidenced by our findings. Serum N-glycan profiles are thus sensitive to the presence of diabetes and MS. Serum N-glycan levels could therefore provide a non-invasive alternative marker for T2DM and MS. PMID:25793407

Bonfigli, Anna Rita; Boemi, Massimo; Olivieri, Fabiola; Ceriello, Antonio; Genovese, Stefano; Spazzafumo, Liana; Borelli, Vincenzo; Bacalini, Maria Giulia; Salvioli, Stefano; Garagnani, Paolo; Dewaele, Sylviane; Libert, Claude; Franceschi, Claudio

2015-01-01

360

Denaturation of human serum albumin under the action of cetyltrimethylammonium bromide according to fluorescence polarization data of protein  

NASA Astrophysics Data System (ADS)

Denaturation of human serum albumin (HSA) under the action of cationic detergent cetyltrimethylammonium bromide (CTAB) is studied at different pH values by estimating the rotational diffusion of protein via fluorescence polarization. The degree of polarization of HSA tryptophan fluorescence, the rotational relaxation time, the rotational diffusion coefficient and the effective Einstein radius of the HSA molecules in solutions with different CTAB concentrations at different pH values are determined. The obtained rotational diffusion parameters of the HSA molecules show that under the action of CTAB, HSA denaturation has a one-stage character and proceeds more intensely and effectively at pH values higher than the p I value of protein (4.7).

Vlasova, I. M.; Zhuravleva, V. V.; Saletskii, A. M.

2012-03-01

361

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

362

The inducible form of heat shock protein 70 in the serum, colon and small intestine of the pig: comparison to conventional stress markers.  

PubMed

Modern rearing conditions may cause stress to pigs. At the cellular level all animals respond to stress by synthesizing heat shock proteins (HSP), which protect cells from injury. The objective of this study was to examine the concentrations of stress-inducible HSP72 in porcine small intestine and colon, known to be stress sensitive tissues, and to compare the findings with HSP72 concentrations in serum and with conventional markers of stress, namely blood lactate and serum cortisol, glucose, free fatty acids and acute phase proteins. HSP72 in the colon correlated with serum HSP72 but there was a negative correlation with carcass weight (growth). The results suggest that the colon may be a significant source of serum HSP72, the concentration of which may reflect changes in the permeability of intestinal epithelium due to stressors, such as transport and handling. PMID:16624719

Sepponen, K; Pösö, A R

2006-05-01

363

The LIM-only protein FHL2 is a serum-inducible transcriptional coactivator of AP-1.  

PubMed

Proteins with LIM domains have been implicated in transcriptional regulation. The four and half LIM domain (FHL) group of LIM-only proteins is composed of five members, some of which have been shown to have intrinsic activation function. Here we show that FHL2 is the only member of the family whose expression is inducible upon serum stimulation in cultured cells. Induction of FHL2 is coordinated in time with the increased levels of two early-response products, the oncoproteins Fos and Jun. FHL2 associates with both Jun and Fos, in vitro and in vivo. The FHL2-Jun interaction requires the Ser-63-Ser-73 JNK phosphoacceptor sites in c-Jun, but not their phosphorylation. FHL2 powerfully stimulates Fos- and Jun-dependent transcription, thereby acting as an inducible coactivator of AP-1 function. Moreover, we show that intracellular localization of FHL2 is controlled by signaling events and a Crm1-dependent active nuclear export mechanism. Thus, FHL2, as an inducible coactivator of AP-1, coordinately participates with Fos and Jun in the early transcriptional response to serum factors. PMID:12644711

Morlon, Aurore; Sassone-Corsi, Paolo

2003-04-01

364

Hevea latex lectin binding protein in C-serum as an anti-latex coagulating factor and its role in a proposed new model for latex coagulation.  

PubMed

A distinct protein specifically recognized by its strong interaction with Hevea latex lectin (HLL) was detected in the aqueous C-serum fraction of centrifuged fresh latex. This C-serum lectin binding protein (CS-HLLBP) exhibited strong inhibition of HLL-induced hemagglutination. The CS-HLLBP was purified to homogeneity by a protocol that included ammonium sulfate fractionation, size exclusion and ion exchange chromatography. The purified CS-HLLBP had a specific HI titer of 0.23microg ml(-1). Its M(r)s analyzed by SDS-PAGE was ca. 40kDa and that by gel filtration was ca. 204kDa. It has a pI value of 4.7, an optimum activity between pH 6 and10 and was heat stable up to 50 degrees C. The HI activity of CS-HLLBP was abolished upon treatment with chitinase. The CS-HLLBP inhibited HLL-induced rubber particle aggregation in a dose dependent manner. A highly positive correlation between CS-HLLBP activity and rubber yield per tapping was found. The correlations for fresh latex (r=0.98, P<0.01) and dry rubber (r=0.95, P<0.01) were both highly significant. This indicated that the CS-HLLBP might be used as a reliable marker for the mass screening of young seedlings to identify and select clones with potential to be superior producers of rubber. A latex anti-coagulating role of the CS-HLLBP is proposed. The findings described in this 3 paper series have been used to propose a new model of rubber latex coagulation that logically describes roles for the newly characterized latex lectin and the two lectin binding proteins. PMID:17983633

Wititsuwannakul, Rapepun; Pasitkul, Piyaporn; Jewtragoon, Pattavuth; Wititsuwannakul, Dhirayos

2008-02-01

365

Orthogonal assembly of a designed ankyrin repeat protein-cytotoxin conjugate with a clickable serum albumin module for half-life extension.  

PubMed

The generation of drug conjugates for safe and effective tumor targeting requires binding proteins tolerant to functionalization by rational engineering. Here, we show that Designed Ankyrin Repeat Proteins (DARPins), a novel class of binding proteins not derived from antibodies, can be used as building blocks for facile orthogonal assembly of bioconjugates for tumor targeting with tailored properties. DARPin Ec1, which targets the Epithelial Cell Adhesion Molecule (EpCAM), was genetically modified with a C-terminal cysteine for conjugation of the small molecule cytotoxin monomethylauristatin F (MMAF). In addition, it was N-terminally functionalized by metabolic introduction of the non-natural amino acid azidohomoalanine to enable linkage of site-specifically dibenzocyclooctyne-modified mouse serum albumin (MSA) for half-life extension using Cu(I)-free click chemistry. The conjugate MSA-Ec1-MMAF was assembled to obtain high yields of a pure and stable drug conjugate as confirmed by various analytical methods and in functional assays. The orthogonality of the assembly led to a defined reaction product and preserved the functional properties of all modules, including EpCAM-specific binding and internalization, FcRn binding mediated by MSA, and cytotoxic potency. Linkage of MMAF to the DARPin increased receptor-specific uptake of the drug while decreasing nonspecific uptake, and further coupling of the conjugate to MSA enhanced this effect. In mice, albumin conjugation increased the serum half-life from 11 min to 17.4 h, resulting in a more than 22-fold increase in the area-under-the-curve (AUC). Our data demonstrate the promise of the DARPin format for facile modular assembly of drug conjugates with improved pharmacokinetic performance for tumor targeting. PMID:24168270

Simon, Manuel; Frey, Raphael; Zangemeister-Wittke, Uwe; Plückthun, Andreas

2013-11-20

366

Potential Protein Toxicity of Synthetic Pigments: Binding of Poncean S to Human Serum Albumin  

Microsoft Academic Search

Using various methods, e.g., spectrophotometry, circular dichroism, and isothermal titration calorimetry, the interaction of poncean S (PS) with human serum albumin (HSA) was characterized at pH 1.81, 3.56, and 7.40 using the spectral correction technique, and Langmuir and Temkin isothermal models. The consistency among results concerning, e.g., binding number, binding energy, and type of binding, showed that ion pair electrostatic

Hong-Wen Gao; Qing Xu; Ling Chen; Shi-Long Wang; Yuan Wang; Ling-Ling Wu; Yuan Yuan

2008-01-01

367

Pre-analytical operating procedures for serum Low Molecular Weight protein profiling  

Microsoft Academic Search

Biological specimen collection and storage are an integral component of serum proteomics research. Although many efforts have been posed to address the effects of pre-analytical procedures, standardized protocols for collection and storage of samples for Low Molecular Weight (LMW) proteome profiling are still needed.Here we report a systematic analysis on the influence of pre-analytical factors [clotting times, temperature and time

Francesco Di Girolamo; Jhessica Alessandroni; Paolo Somma; Fiorella Guadagni

2010-01-01

368

Capillary electrophoresis for clinical problem solving: analysis of urinary diagnostic metabolites and serum proteins  

Microsoft Academic Search

Many clinical laboratories employ gas chromatography-mass spectrometry (GC-MS) and high-performance liquid chromatography (HPLC) to detect abnormal compounds occurinng in urine and serum due to disease. The methods, particularly GC-MS, often require laborious sample pre-treatment, and separation times may exceed an hour. We describe the use of capillary electrophoresis (CE) equipped a with a diode-array detector in an attempt to improve

Egil Jellum; Herman Dollekamp; Cynthia Blessum

1996-01-01

369

Intuitive, but not simple: including explicit water molecules in protein-protein docking simulations improves model quality.  

PubMed

Characterizing the nature of interaction between proteins that have not been experimentally cocrystallized requires a computational docking approach that can successfully predict the spatial conformation adopted in the complex. In this work, the Hydropathic INTeractions (HINT) force field model was used for scoring docked models in a data set of 30 high-resolution crystallographically characterized "dry" protein-protein complexes and was shown to reliably identify native-like models. However, most current protein-protein docking algorithms fail to explicitly account for water molecules involved in bridging interactions that mediate and stabilize the association of the protein partners, so we used HINT to illuminate the physical and chemical properties of bridging waters and account for their energetic stabilizing contributions. The HINT water Relevance metric identified the "truly" bridging waters at the 30 protein-protein interfaces and we utilized them in "solvated" docking by manually inserting them into the input files for the rigid body ZDOCK program. By accounting for these interfacial waters, a statistically significant improvement of ?24% in the average hit-count within the top-10 predictions the protein-protein dataset was seen, compared to standard "dry" docking. The results also show scoring improvement, with medium and high accuracy models ranking much better than incorrect ones. These improvements can be attributed to the physical presence of water molecules that alter surface properties and better represent native shape and hydropathic complementarity between interacting partners, with concomitantly more accurate native-like structure predictions. PMID:24214407

Parikh, Hardik I; Kellogg, Glen E

2014-06-01

370

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

371

Bronchoalveolar Lavage Fluid and Serum Canine Surfactant Protein A Concentrations in Dogs with Chronic Cough by Bronchial and Interstitial Lung Diseases  

PubMed Central

ABSTRACT We measured bronchoalveolar lavage fluid (BALF) and serum canine surfactant protein (cSP)-A concentrations in dogs with chronic cough. There were no significant differences between bronchial and interstitial lung diseases in BALF cSP-A concentrations. However, serum cSP-A concentrations in dogs with the interstitial lung disease as diffuse panbronchiolitis and idiopathic pulmonary fibrosis were significantly higher than those in dogs with the bronchial disease as chronic bronchitis. These results suggest that serum cSP-A concentrations may be a useful and noninvasive biomarker to understand the existence of interstitial lung damage in dogs with chronic cough. PMID:24366151

YAMAYA, Yoshiki; SUZUKI, Kazuyuki; WATARI, Toshihiro; ASANO, Ryuji

2013-01-01

372

SNAP family of NSF attachment proteins includes a brain-specific isoform  

Microsoft Academic Search

THE soluble NSF attachment proteins (SNAPs) enable N-ethyl-maleimide-sensitive fusion protein (NSF) to bind to target mem-branes1-4. Here we report the cloning and sequencing of com-plementary DNAs encoding alpha-, beta- and gamma-SNAPs. Two of these proteins, alpha and gamma, are found in a wide range of tissues, and act synergistically in intra-Golgi transport. The third, beta, is a brain-specific isoform of

Sidney W. Whiteheart; Irene C. Griff; Michael Brunner; Douglas O. Clary; Thomas Mayer; Susan A. Buhrow; James E. Rothman

1993-01-01

373

Accurate Quantification of Cardiovascular Biomarkers in Serum Using Protein Standard Absolute Quantification (PSAQ™) and Selected Reaction Monitoring*  

PubMed Central

Development of new biomarkers needs to be significantly accelerated to improve diagnostic, prognostic, and toxicity monitoring as well as therapeutic follow-up. Biomarker evaluation is the main bottleneck in this development process. Selected Reaction Monitoring (SRM) combined with stable isotope dilution has emerged as a promising option to speed this step, particularly because of its multiplexing capacities. However, analytical variabilities because of upstream sample handling or incomplete trypsin digestion still need to be resolved. In 2007, we developed the PSAQ™ method (Protein Standard Absolute Quantification), which uses full-length isotope-labeled protein standards to quantify target proteins. In the present study we used clinically validated cardiovascular biomarkers (LDH-B, CKMB, myoglobin, and troponin I) to demonstrate that the combination of PSAQ and SRM (PSAQ-SRM) allows highly accurate biomarker quantification in serum samples. A multiplex PSAQ-SRM assay was used to quantify these biomarkers in clinical samples from myocardial infarction patients. Good correlation between PSAQ-SRM and ELISA assay results was found and demonstrated the consistency between these analytical approaches. Thus, PSAQ-SRM has the capacity to improve both accuracy and reproducibility in protein analysis. This will be a major contribution to efficient biomarker development strategies. PMID:22080464

Huillet, Céline; Adrait, Annie; Lebert, Dorothée; Picard, Guillaume; Trauchessec, Mathieu; Louwagie, Mathilde; Dupuis, Alain; Hittinger, Luc; Ghaleh, Bijan; Le Corvoisier, Philippe; Jaquinod, Michel; Garin, Jérôme; Bruley, Christophe; Brun, Virginie

2012-01-01

374

High-performance liquid chromatography of proteins: purification of alpha-fetoprotein from fetal calf serum.  

PubMed

High-performance liquid chromatography was utilized for the purification of bovine alpha-fetoprotein (BAFP) from fetal calf serum (FCS). An initial step in the purification involved absorption of charcoal delipidated FCS on Cibacron Blue F3GA gel. The Cibacron Blue pre-purified FCS was then chromatographed on a Polyanion SI weak anion-exchange column. The BAFP isolated had a purity of greater than 93% with an overall yield of 48% from FCS. The procedure was applicable for semi-preparative scale purification of BAFP. PMID:6209297

Wong, L T; Hsia, J C

1984-09-14

375

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

E-print Network

in the mammalian body. These proteins are essential constituents of food, drugs and cosmetics, and their dynamics, metabo- lites, lipophilic compounds, hormones and drugs, including anesthetics and anti-coagulants.1

376

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

377

Soybean protein lowers serum cholesterol levels in hamsters: Effect of debittered undigested fraction  

Microsoft Academic Search

the undigested fraction (UDF) of soybean protein exerts a marked hypocholesterolemic effect in relation to soybean protein (SOY) in rats. The present study was conducted to confirm whether UDF was effective in hamsters as in rats in combination with different fat sources, either perilla oil (PER) or safflower oil (SAF). Because the hamster, unlike the rat, disliked the bitter taste

Marites Gatchalian-Yee; Yoko Arimura; Eiko Ochiai; Koji Yamada; Michihiro Sugano

1997-01-01

378

Serum Tau Protein Level as a Marker of Axonal Damage in Acute Ischemic Stroke  

Microsoft Academic Search

Biochemical markers of brain damage, e.g. ischemic stroke, should reflect the volume of irreversibly damaged brain parenchyma and the clinical outcome in a single patient in order to allow estimation of prognosis at an early stage. Tau protein, which derives predominantly from neurons and axons, is elevated in the cerebrospinal fluid of patients with neurodegenerative disease. This makes tau protein

Andreas Bitsch; Claudia Horn; Yvonne Kemmling; Maria Seipelt; Uwe Hellenbrand; Michael Stiefel; Barbara Ciesielczyk; Lukas Cepek; Erik Bahn; Peter Ratzka; Hilmar Prange; Markus Otto

2002-01-01

379

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

380

Identification of a locus modulating serum C-reactive protein levels on chromosome 5p15  

PubMed Central

Objective: Individual propensity to chronic, low–grade inflammation – a determinant of atherosclerosis – is in part under the control of genetic factors. To identify genes involved in this modulation, we performed a 10 cM genome screen for linkage with plasma C-reactive protein in 38 extended families including 317 non-diabetic and 177 type 2 diabetic family members (2,547 relative pairs). Methods and results: In a variance component analysis, heritability of CRP values was significant (h2=0.39, p<0.0001). This effect was independent of BMI and was present in both diabetic (h2=0.42, p=0.003) and non-diabetic (h2=0.34, p=0.0015) relatives. The strongest evidence of linkage with CRP was on chromosome 5p15, where the LOD score reached genome-wide significance (LOD=3.41, genome-wide p=0.013). Both diabetic and non-diabetic family members contributed to linkage at this location. Smaller linkage peaks were detected on chromosomes 5q35 (LOD=1.35) and 17p11 (LOD=1.33). When the analysis was restricted to diabetic family members, another peak of moderate intensity (LOD=2.17) was evident at 3p21. Conclusions: A major gene influencing CRP levels appears to be located on chromosome 5p15, with an effect that is independent of diabetes. Another gene on 3p21 may control CRP variation but only in the presence of a diabetic or insulin-resistant environment. PMID:17343862

Keenan, Hillary A.; Poznik, G. David; Varo, Nerea; Schneider, Jennifer; Almasy, Laura; Warram, James H.; Duggirala, Ravindranath; Schoenbeck, Uwe; Krolewski, Andrzej S.; Doria, Alessandro

2008-01-01

381

PIAS1 activates the expression of smooth muscle cell differentiation marker genes by interacting with serum response factor and class I basic helix-loop-helix proteins.  

PubMed

Although a critical component of vascular disease is modulation of the differentiated state of vascular smooth muscle cells (SMC), the mechanisms governing SMC differentiation are relatively poorly understood. We have previously shown that E-boxes and the ubiquitously expressed class I basic helix-loop-helix (bHLH) proteins, including E2-2 and E12, are important in regulation of the SMC differentiation marker gene, the SM alpha-actin gene. The aim of the present study was to identify proteins that bind to class I bHLH proteins in SMC and modulate transcriptional regulation of SMC differentiation marker genes. Herein we report that members of the protein inhibitor of activated STAT (PIAS) family interact with class I bHLH factors as well as serum response factor (SRF). PIAS1 interacted with E2-2 and E12 based on yeast two-hybrid screens, mammalian two-hybrid assays, and/or coimmunoprecipitation assays. Overexpression of PIAS1 significantly activated the SM alpha-actin promoter and mRNA expression, as well as SM myosin heavy chain and SM22alpha, whereas a small interfering RNA for PIAS1 decreased activity of these promoters, as well as endogenous mRNA expression, and SRF binding to SM alpha-actin promoter within intact chromatin in cultured SMC. Of significance, PIAS1 bound to SRF and activated SM alpha-actin promoter expression in wild-type but not SRF(-/-) embryonic stem cells. These results provide novel evidence that PIAS1 modulates transcriptional activation of SMC marker genes through cooperative interactions with both SRF and class I bHLH proteins. PMID:16135793

Kawai-Kowase, Keiko; Kumar, Meena S; Hoofnagle, Mark H; Yoshida, Tadashi; Owens, Gary K

2005-09-01

382

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

383

Detection of hepatitis C virus core protein in serum by atomic force microscopy combined with mass spectrometry  

PubMed Central

A method for detection and identification of core antigen of hepatitis C virus (HCVcoreAg)-containing particles in the serum was proposed, with due account taken of the interactions of proteotypic peptides with Na+, K+, and Cl? ions. The method is based on a combination of reversible biospecific atomic force microscopy (AFM)-fishing and mass spectrometry (MS). AFM-fishing enables concentration, detection, and counting of protein complexes captured on the AFM chip surface, with their subsequent MS identification. Biospecific AFM-fishing of HCVcoreAg-containing particles from serum samples was carried out using AFM chips with immobilized antibodies against HCVcoreAg (HCVcoreAgim). Formation of complexes between anti-HCVcoreAgim and HCVcoreAg-containing particles on the AFM chip surface during the fishing process was demonstrated. These complexes were registered and counted by AFM. Further MS analysis allowed reliable identification of HCVcoreAg within the complexes formed on the AFM chip surface. It was shown that MS data processing, with account taken of the interactions between HCVcoreAg peptides and Na+, K+ cations, and Cl? anions, allows an increase in the number of peptides identified.

Ivanov, Yuri D; Kaysheva, Anna L; Frantsuzov, Pavel A; Pleshakova, Tatyana O; Krohin, Nikolay V; Izotov, Alexander A; Shumov, Ivan D; Uchaikin, Vasiliy F; Konev, Vladimir A; Ziborov, Vadim S; Archakov, Alexander I

2015-01-01

384

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

385

Discovery of serum protein biomarkers in rheumatoid arthritis using MALDI-TOF-MS combined with magnetic beads.  

PubMed

The aim of this study was to discover potential biomarkers for rheumatoid arthritis (RA) using Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry combined with magnetic beads. Proteomic fingerprint technology combining magnetic beads with MALDI-TOF-MS was used to profile and compare the proteomes in serum samples from 60 patients with RA, 35 patients with osteoarthritis and 36 healthy controls. The proteomic pattern associated with RA was identified by Biomarker Patterns Software. Model of biomarkers was constructed and evaluated through the Biomarker Patterns Software. A total of 33 discriminative peaks were identified to be related with RA, in which the 5 peaks with the mass-charge ratio (m/z) peaks at 15,715.5, 7,771.4, 8,959.4, 8,469.8 and 8,710.8 Da were used to construct a model for the diagnosis of RA by pattern recognition software. The blind testing data indicated a sensitivity of 86.7% and a specificity of 90.0% in RA diagnosis. These results demonstrated that potential protein biomarkers for RA could be discovered in serum by MALDI-TOF-MS combined with WCX magnetic beads. The diagnosis mode tree based on the five candidate biomarkers could provide a powerful and reliable diagnostic method for RA with high sensitivity and specificity. PMID:21922190

Zhang, Xiaoxue; Yuan, Zhaolin; Shen, Bo; Zhu, Min; Liu, Chibo; Xu, Wei

2012-09-01