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Protein electrophoresis - serum  


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


A mouse hepatoma cell line which secretes several serum proteins including albumin and alpha-foetoprotein.  


A permanent cell line (BW) was established from a transplantable mouse hepatoma, BW7756, which produces alpha-foetoprotein (AFP). Three clones were isolated from the uncloned culture: BW1, BW2 and BWTG3. The cells of the latter clone, which was isolated after selection in the presence of thioguanine, are deficient in the enzyme hypoxanthine-guanine-phosphoribosyl transferase. Both BW1 and BWTG3 cells have mean chromosome number of 64 (60 telocentric and 4 metacentric chromosomes). All three clones secrete at least five serum proteins into the culture medium: albumin, AFP, and alpha 2 globulin, transferrin and C3, the third component of complement. The approximate rate of albumin secretion by BW1 and BWTG3 cells is 10 mug/24 h/10(6) cells. Both albumin and AFP can easily be detected in cell extracts. The simultaneous production of AFP and a hepatocyte specific marker (albumin) by cloned hepatoma cells show that the production of AFP by the tumour is due to the tumoural hepatocytes themselves. PMID:52568

Szpirer, C; Szpirer, J



Human serum amyloid P is a multispecific adhesive protein whose ligands include 6-phosphorylated mannose and the 3-sulphated saccharides galactose, N-acetylgalactosamine and glucuronic acid.  

PubMed Central

Carbohydrate recognition by amyloid P component from human serum has been investigated by binding experiments using several glycosaminoglycans, polysaccharides and a series of structurally defined neoglycolipids and natural glycolipids. Two novel classes of carbohydrate ligands have been identified. The first is 6-phosphorylated mannose as found on lysosomal hydrolases, and the second is the 3-sulphated saccharides galactose, N-acetyl-galactosamine and glucuronic acid as found on sulphatide and other acidic glycolipids that occur in neural or kidney tissues or on subpopulations of lymphocytes. Binding to mannose-6-phosphate containing molecules and inhibition of binding by free mannose-6-phosphate and fructose-1-phosphate are features shared with mannose-6-phosphate receptors involved in trafficking of lysosomal enzymes. However, only amyloid P binding is inhibited by galactose-6-phosphate, mannose-1-phosphate and glucose-6-phosphate. These findings strengthen the possibility that amyloid P protein has a central role in amyloidogenic processes: first in formation of focal concentrations of lysosomal enzymes including proteases that generate fibril-forming peptides from amyloidogenic proteins, and second in formation of multicomponent complexes that include sulphoglycolipids as well as glycosaminoglycans. The evidence that binding to all of the acidic ligands involves the same polypeptide domain on amyloid P protein, and inhibition data using diffusible, phosphorylated monosaccharides, is potentially important leads to novel drug designs aimed at preventing or even reversing amyloid deposition processes without interference with essential lysosomal trafficking pathways. Images

Loveless, R W; Floyd-O'Sullivan, G; Raynes, J G; Yuen, C T; Feizi, T



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


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

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



Serum heme-albumin: an allosteric protein.  


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

Ascenzi, Paolo; Fasano, Mauro



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.


Postpartum Changes in Milk Serum Protein Fractions  

Microsoft Academic Search

Polyaerylamide gel disc electrophoresis was used to study changes in the relative concentration of the fi-lactoglobulin, a- laetalbumin, blood serum albumin, and im- mune globulin in milk serum during the first 21 days post-partum. All four protein fractions were found to be concentrated in cotostrum serum at parturition, with im- mune globulin being a major fraction. A graphic analysis of

R. M. Porter; H. R. Conrad



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



Human serum protein fractionation by gel filtration  

PubMed Central

The development of a quantitative immunological technique using polyvalent antiserum permits a more logical approach to the fractionation of complex protein mixtures. In this study whole serum was separated by conventional gel filtration and the fractions obtained were analysed. This demonstrates over 60 immunologically distinct serum proteins. Because the current terminology is inadequate to describe this number of proteins, a temporary numerical nomenclature has been used. ImagesPLATE 4PLATE 5PLATE 1PLATE 2PLATE 3

Freeman, T.; Smith, J.



(PCG) Protein Crystal Growth Horse Serum Albumin  

NASA Technical Reports Server (NTRS)

Horse Serum Albumin crystals grown during the USML-1 (STS-50) mission's Protein Crystal Growth Glovebox Experiment. These crystals were grown using a vapor diffusion technique at 22 degrees C. The crystals were allowed to grow for nine days while in orbit. Crystals of 1.0 mm in length were produced. The most abundant blood serum protein, regulates blood pressure and transports ions, metabolites, and therapeutic drugs. Principal Investigator was Edward Meehan.



Proteomic evaluation of sheep serum proteins  

PubMed Central

Background The applications of proteomic strategies to ovine medicine remain limited. The definition of serum proteome may be a good tool to identify useful protein biomarkers for recognising sub-clinical conditions and overt disease in sheep. Findings from bovine species are often directly translated for use in ovine medicine. In order to characterize normal protein patterns and improve knowledge of molecular species-specific characteristics, we generated a two-dimensional reference map of sheep serum. The possible application of this approach was tested by analysing serum protein patterns in ewes with mild broncho-pulmonary disease, which is very common in sheep and in the peripartum period which is a stressful time, with a high incidence of infectious and parasitic diseases. Results This study generated the first reference 2-DE maps of sheep serum. Overall, 250 protein spots were analyzed, and 138 identified. Compared with healthy sheep, serum protein profiles of animals with rhino-tracheo-bronchitis showed a significant decrease in protein spots identified as transthyretin, apolipoprotein A1 and a significant increase in spots identified as haptoglobin, endopin 1b and alpha1B glycoprotein. In the peripartum period, haptoglobin, alpha-1-acid glycoprotein, apolipoprotein A1 levels rose, while transthyretin content dropped. Conclusions This study describes applications of proteomics in putative biomarker discovery for early diagnosis as well as for monitoring the physiological and metabolic situations critical for ovine welfare.



Differential serum protein markers and the clinical severity of asthma  

PubMed Central

Background Asthma is a heterogeneous disease characterized by different clinical phenotypes and the involvement of multiple inflammatory pathways. During airway inflammation, many cytokines and chemokines are released and some are detectable in the sera. Objective Serum chemokines and cytokines, involved in airway inflammation in asthma patients, were investigated. Methods A total of 191 asthma patients were classified by hierarchical cluster analysis, including the following parameters: forced expiratory volume in 1 second (FEV1), eosinophil cationic protein (ECP) serum levels, blood eosinophils, Junipers asthma symptom score, and the change in FEV1, ECP serum levels, and blood eosinophils after 3 weeks of asthma therapy. Serum proteins were measured by multiplex analysis. Receiver operating characteristic (ROC) curves were used to evaluate the validity of serum proteins for discriminating between asthma clusters. Results Classification of asthma patients identified one cluster with high ECP serum levels, increased blood eosinophils, low FEV1 values, and good FEV1 improvement in response to asthma therapy (n=60) and one cluster with low ECP serum levels, low numbers of blood eosinophils, higher FEV1 values, and no FEV1 improvement in response to asthma therapy (n=131). Serum interleukin (IL)-8, eotaxin, vascular endothelial growth factor (VEGF), cutaneous T-cell-attracting chemokine (CTACK), growth-related oncogene (GRO)-?, and hepatocyte growth factor (HGF) were significantly different between the two clusters of asthma patients. ROC analysis for serum proteins calculated a sensitivity of 55.9% and specificity of 75.8% for discriminating between them. Conclusion Serum cytokine and chemokine levels might be predictors for the severity of asthmatic inflammation, asthma control, and response to therapy, and therefore might be useful for treatment optimization.

Meyer, Norbert; Nuss, Sarah Janine; Rothe, Thomas; Siebenhuner, Alexander; Akdis, Cezmi A; Menz, Gunter



Biomarker Amplification by Serum Carrier Protein Binding  

PubMed Central

Mass spectroscopic analysis of the low molecular mass (LMM) range of the serum/plasma proteome is a rapidly emerging frontier for biomarker discovery. This study examined the proportion of LMM biomarkers, which are bound to circulating carrier proteins. Mass spectroscopic analysis of human serum following molecular mass fractionation, demonstrated that the majority of LMM biomarkers exist bound to carrier proteins. Moreover, the pattern of LMM biomarkers bound specifically to albumin is distinct from those bound to non-albumin carriers. Prominent SELDI-TOF ionic species (m/z 6631.7043) identified to correlate with the presence of ovarian cancer were amplified by albumin capture. Several insights emerged: a) Accumulation of LMM biomarkers on circulating carrier proteins greatly amplifies the total serum/plasma concentration of the measurable biomarker, b) The total serum/plasma biomarker concentration is largely determined by the carrier protein clearance rate, not the unbound biomarker clearance rate itself, and c) Examination of the LMM species bound to a specific carrier protein may contain important diagnostic information. These findings shift the focus of biomarker detection to the carrier protein and its biomarker content.

Mehta, Arpita I.; Ross, Sally; Lowenthal, Mark S.; Fusaro, Vincent; Fishman, David A.; Petricoin, Emanuel F.; Liotta, Lance A.



(PCG) Protein Crystal Growth Human Serum Albumin  

NASA Technical Reports Server (NTRS)

(PCG) Protein Crystal Growth Human Serum Albumin. Contributes to many transport and regulatory processes and has multifunctional binding properties which range from various metals, to fatty acids, hormones, and a wide spectrum of therapeutic drugs. The most abundant protein of the circulatory system. It binds and transports an incredible variety of biological and pharmaceutical ligands throughout the blood stream. Principal Investigator on STS-26 was Larry DeLucas.



Quantitating metabolites in protein precipitated serum using NMR spectroscopy.  


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



A proteomic reference map for pig serum proteins as a prerequisite for diagnostic applications  

Microsoft Academic Search

A reference protein map for pig serum was set up using two-dimensional electrophoresis (2-DE). Thirty-nine protein chains or spots deriving from 26 different proteins were identified by immunological and mass spectrometric methods. Thus, the positions of most medium to higher abundance serum proteins could be determined on the 2-DE gels. The plasma protein fibrinogen was also included in our study.

Ingrid Miller; Robin Wait; Wolfgang Sipos; Manfred Gemeiner



Anion exchange fractionation of serum proteins versus albumin elimination  

Microsoft Academic Search

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

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



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

PubMed Central

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 the host response. Methods Eighty-four antibodies, targeting a wide range of serum proteins, were spotted onto nitrocellulose-coated microscope slides. The abundances of the corresponding proteins were measured in 80 serum samples, from 24 newly diagnosed subjects with lung cancer, 24 healthy controls, and 32 subjects with chronic obstructive pulmonary disease (COPD). Two-color rolling-circle amplification was used to measure protein abundance. Results Seven of the 84 antibodies gave a significant difference (p < 0.01) for the lung cancer patients as compared to healthy controls, as well as compared to COPD patients. Proteins that exhibited higher abundances in the lung cancer samples relative to the control samples included C-reactive protein (CRP; a 13.3 fold increase), serum amyloid A (SAA; a 2.0 fold increase), mucin 1 and ?-1-antitrypsin (1.4 fold increases). The increased expression levels of CRP and SAA were validated by Western blot analysis. Leave-one-out cross-validation was used to construct Diagonal Linear Discriminant Analysis (DLDA) classifiers. At a cutoff where all 56 of the non-tumor samples were correctly classified, 15/24 lung tumor patient sera were correctly classified. Conclusion Our results suggest that a distinctive serum protein profile involving abundant proteins may be observed in lung cancer patients relative to healthy subjects or patients with chronic disease and may have utility as part of strategies for detecting lung cancer.

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



Serum alkaline phosphatase levels associate with elevated serum C-reactive protein in chronic kidney disease  

Microsoft Academic Search

High serum alkaline phosphatase concentrations are associated with elevated serum C-reactive protein (CRP) levels in the general population. To examine whether this association is independent of serum vitamin D levels or modified in chronic kidney disease (CKD), we determined if such associations exist using data from the National Health and Nutrition Examination Survey III of 14,420 adult participants in which

Sriharsha Damera; Kalani L Raphael; Bradley C Baird; Alfred K Cheung; Tom Greene; Srinivasan Beddhu



Sensing of Proteins in Human Serum using Nanoparticle-Green Fluorescent Protein Conjugates  

PubMed Central

There is a direct correlation between protein levels and disease states in human serum making it an attractive target for sensors and diagnostics. However this is made challenging because serum features more than 20,000 proteins with an overall protein content of greater than 1 mM. Here we report a hybrid synthetic-biomolecule based sensor that uses green fluorescent protein-nanoparticle arrays to detect proteins at biorelevant concentrations in both buffer and human serum. Distinct and reproducible fluorescence response patterns were obtained from five serum proteins (human serum albumin, immunoglobulin G, transferrin, fibrinogen and ?-antitrypsin) in buffer and when spiked into human serum. Using linear discriminant analysis we identified these proteins with an identification accuracy of 100% in buffer and 97% in human serum. The arrays were also able to discriminate between different concentrations of the same protein as well as a mixture of different proteins in human serum.




Characterization of Brevin, a Serum Protein That Shortens Actin Filaments  

Microsoft Academic Search

We have purified a protein from rabbit serum with a molecular weight of 90,000 that inhibits the polymerization of actin measured viscometrically and that we have named ``brevin'' (from the Latin breviare, to shorten). From the extent of purification, we estimate that this inhibitor constitutes 0.3% of the total protein in plasma and serum. Brevin is also present in sera

David A. Harris; James H. Schwartz




PubMed Central

1. By the technique of quantitative plasmapheresis the effects of single proteins in artificial synthetic diets were studied with respect to their value in promoting the regeneration of serum protein. 2. The ratio of (a) the amount of serum protein per week removed by bleeding above that regenerated by the dog when eating the protein-free diet, to (b) the dietary protein increment (i.e., above that required for nitrogen equilibrium) was termed the potency ratio. The results indicated that serum protein was slightly superior to casein and lactalbumin in promoting the regeneration of serum protein. However, the respective potency ratios, varying from approximately 0.51 to 0.36, were comparable and not widely divergent as those reported by others. It was concluded that, whereas in some individuals dietary proteins may be able to produce a significant increase in the serum protein concentration, the potency ratios are not sufficiently different to warrant the administration of any one protein in preference to another. 3. The inhibitory effect of the basal protein-free diet with respect to serum protein regeneration in the dog was also demonstrated by the inability of the protein concentration to attain the normal level in spite of discontinued plasmapheresis. However, a subsequent fasting period resulted in a progressive rise in the serum protein concentration until the normal value was approximated. These observations are interpreted as indicating that the products of tissue protein catabolism can be utilized in the formation of new serum protein. 4. The experimental production of what seems to be an inhibition of the serum protein regenerating mechanism was described. This observation together with the hypothetical evidence presented by Bloomfield (17) and Weech and his associates (9) suggests that the most profitable line of approach to solution of the problem of hypoproteinemia lies not so much in the evaluation of dietary factors but in finding a way for stimulating internally the serum protein regenerating mechanism, which seems to involve in some manner the capacity of the tissue to furnish protein for the needs of the plasma. 5. A hypothesis explaining the mechanisms responsible for serum protein formation was presented and the experimental support for it discussed. The rôle of tissue protein catabolism in this function was emphasized.

Melnick, Daniel; Cowgill, George R.; Burack, Ethel



Serum acute phase proteins and swine health status  

PubMed Central

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

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



Serum tau protein level serves as a predictive factor for neurological prognosis in neonatal asphyxia.  


Background: Tau protein is a microtubule-associated protein that is present in axons. Elevated tau protein levels in cerebrospinal fluid or serum are associated with several central nervous system diseases and can indicate neuronal injury. Objective: In the present study, we measured and then compared serum tau protein levels between infants with neonatal asphyxia and control subjects. We examined these data to investigate the correlation between serum tau protein levels and neurological outcomes after neonatal asphyxia. Patients and methods: Serum tau protein levels were determined by an enzyme-linked immunosorbent assay in 19 neonates with neonatal asphyxia. Of these 19 neonates, 3 had severe spastic tetraplegia, and 1 had west syndrome. A group of 19 unaffected neonates was included in the study as a control group. Results: Serum tau protein levels on postnatal day 3 were significantly higher in the poor outcome group than those in the good outcome (p=0.010) and control groups (p=0.006). On postnatal day 7, serum tau protein levels again were significantly higher in the poor outcome group than those in the good outcome (p=0.007) and control groups (p=0.006). Conclusions: The present findings indicate serum tau protein levels measured on postnatal days 3 and 7 can predict neurological prognosis following neonatal asphyxia. PMID:24268747

Takahashi, Kazumasa; Hasegawa, Shunji; Maeba, Shinji; Fukunaga, Shinnosuke; Motoyama, Masashi; Hamano, Hiroki; Ichiyama, Takashi



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


Characterization of an Anticryptococcal Protein Isolated from Human Serum  

PubMed Central

Human serum at low concentrations inhibits the growth of Cryptococcus neoformans in vitro. Fractionation of serum yielded a purified inhibitory protein with a molecular mass of ?81.8 kDa, a pI of ?6.2, and an amino acid sequence that matched that of human transferrin. The inhibitory activity and that of apotransferrin and 5% human serum were reversed by 10 ?M freshly prepared FeCl3.

Sridhar, Sridevi; Ahluwalia, Mala; Brummer, Elmer; Stevens, David A.



Grizzly bear corticosteroid binding globulin: Cloning and serum protein expression  

Microsoft Academic Search

Serum corticosteroid levels are routinely measured as markers of stress in wild animals. However, corticosteroid levels rise rapidly in response to the acute stress of capture and restraint for sampling, limiting its use as an indicator of chronic stress. We hypothesized that serum corticosteroid binding globulin (CBG), the primary transport protein for corticosteroids in circulation, may be a better marker

Brian A. Chow; Jason Hamilton; Derek Alsop; Marc R. L. Cattet; Gordon Stenhouse; Mathilakath M. Vijayan



Binding between lead ions and the high-abundance serum proteins.  


The interaction between three of the most abundant bovine serum proteins (serum albumin, transferrin and IgG) with Pb(2+) was investigated using electrochemistry. The data was used to construct a new theoretical model of Pb(2+) binding to the high-abundance serum proteins under non-ideal conditions. The binding constants (?) of Pb(2+) to the individual proteins and a mixture of proteins were measured according to a new theoretical equation (non-ideal state) as well as the McGhee-Von Hippel equation (ideal state). Differences between the models suggested that the ? values obtained using the non-ideal state model was more realistic. Protein-protein interactions and micro-environmental influences affected binding between Pb(2+) and the high-abundance serum proteins. We included a micro-environmental influence factor for the model (Fm), which accurately quantified the effect of micro-environment of the proteome of Pb(2+) binding with the serum proteins. This research provides a useful reference of theoretical and experimental work regarding heavy-metal binding interactions with serum proteins. PMID:25048942

Guo, Ming; He, Ling; Strong, P J; Wang, Hailong




Microsoft Academic Search

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

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



Raised serum S100B protein levels in neuropsychiatric lupus  

Microsoft Academic Search

Objective: To test serum S100B protein levels in patients with and without neuropsychiatric systemic lupus erythematosus (NPSLE) and controls.Methods: 87 patients with SLE, 23 with and 64 without neuropsychiatric involvement, and 25 control subjects were prospectively evaluated. NPSLE diagnosis was made according to the American College of Rheumatology nomenclature and case definitions for neuropsychiatric lupus syndromes. Serum S100B protein levels

C B Schenatto; R M Xavier; M Bredemeier; L V C Portela; A B L Tort; T L Dedavid e Silva; D O Souza; J C T Brenol



Spectroscopic imaging of serum proteins using quantum cascade lasers.  


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/cm˛ 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/cm˛. 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



Spectroscopic imaging of serum proteins using quantum cascade lasers  

NASA Astrophysics Data System (ADS)

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.

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



Usual Intake of Total protein foods including beans and peas

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%


Inactivation of Anthracyclines by Serum Heme Proteins  

Microsoft Academic Search

We have previously shown that the anticancer agent doxorubicin undergoes oxidation and inactivation when exposed to myeloperoxidase-containing human leukemia HL-60 cells, or to isolated myeloperoxidase, in the presence of hydrogen peroxide and nitrite. In the current study we report that commercial fetal bovine serum (FBS) alone oxidizes doxorubicin in the presence of hydrogen peroxide and that nitrite accelerates this oxidation.

Brett A. Wagner; Lynn M. Teesch; Garry R. Buettner; Bradley E. Britigan; C. Patrick Burns; Krzysztof J. Reszka



Interaction of Serum Proteins with Surface of Hemodialysis Fiber Membranes  

NASA Astrophysics Data System (ADS)

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

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



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



Relationship of Blood Serum Protein and Protein Fractions to Milk Constituents and Temperature-Season  

Microsoft Academic Search

Blood serum protein and protein frac- tions were determined in 265 samples from 62 lactating Holstein cows over one calendar year. Temperature-seasonal changes (coo), optimum, hot) were re- sponsible for significant linear and quad- ratic fluctuation in serum protein frac- tions. Serum albumin, fl-globulin, a2- globulin, and the A:G ratio significantly decreased as seasons progressed from cool to hot. Further,

J. D. Roussel; K. L. Koonce; M. A. Pinero



Erythropoietin binding protein from mammalian serum  


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

Clemons, Gisela K. (Berkeley, CA)



Erythropoietin binding protein from mammalian serum  


Purified mammalian erythropoietin binding-protein is disclosed, and its isolation, identification, characterization, purification, and immunoassay are described. The erythropoietin binding protein can be used for regulation of erythropoiesis by regulating levels and half-life of erythropoietin. A diagnostic kit for determination of level of erythropoietin binding protein is also described. 11 figs.

Clemons, G.K.



Influence of serum protein, serum albumin concentrations and dose on midazolam anaesthesia induction times  

Microsoft Academic Search

Individual variation occurs in time to induction of anaesthesia with intravenous drugs. Less free drug is available to cross\\u000a the blood-brain barrier when the drug is highly protein bound. Since this may prolong time to sleep, we correlated the induction\\u000a time, serum albumin and total protein concentrations, and doses of midazolam, which is a highly protein bound intravenous\\u000a anaesthetic. There

J. G. Ręves; Philippa Newfield; Lloyd R. Smith



Association of Leucogenenol with Specific Proteins in Serum.  

National Technical Information Service (NTIS)

Determination of the concentration of leucogenenol in the IgM, IgG, albumin, and low molecular weight protein fractions of serum demonstrated that leucogenenol is found only in protein fractions of molecular weights in the range of IgM and IgG. Determinat...

F. A. H. Rice B. R. Griffin P. D. Dass



Sea lamprey (Petromyzon marinus) contain four developmentally regulated serum thyroid hormone distributor proteins.  


Thyroid hormones (THs) are very lipophilic molecules which require a distribution network for efficient transport in serum. Despite observations that THs function in a wide variety of processes, including aspects of fish development (i.e., flat fish metamorphosis and smoltification), the proteins responsible for TH distribution in fish serum remain poorly studied. We chose to investigate the serum TH distributor proteins (THDPs) in lampreys. As one of only two extant agnathans, data on lamprey THDPs may offer new insights into the evolution of the vertebrate TH distribution network and serum proteins in general. Moreover, lampreys appear to contradict the vertebrate model of an increase in TH concentrations initiating and driving vertebrate metamorphosis. We show for the first time that sea lamprey serum contains at least four THDPs and that their presence in serum is temporally regulated throughout the life cycle. The albumin, glycoprotein AS is the dominant THDP present in the sera of larval and metamorphosing sea lamprey. In stage seven of metamorphosis, three additional THDPs appear, including the albumin, glycoprotein SDS-1; the glycolipoprotein CB-III; and an unidentified low molecular weight protein temporarily named Spot-5. The sera of parasitic and upstream migrant sea lampreys lack AS; their serum THDPs are SDS-1, CB-III, and Spot-5. Our data indicate that despite the change in type and number of THDPs, the overall total TH binding capacity of sea lamprey serum remains fairly stable until stage 7 of metamorphosis when a only modest decrease in total binding capacity is observed. Collectively these data indicate that the decline in serum TH concentrations observed during lamprey metamorphosis is not a consequence of a reduction in the distribution and storage capacity of the serum. PMID:21163261

Gross, Tianna Natalia; Manzon, Richard Giuseppe



Complement regulator-acquiring surface protein 1 imparts resistance to human serum in Borrelia burgdorferi.  


Factor H and factor H-like protein 1 (FH/FHL-1) are soluble serum proteins that negatively regulate the alternative pathway of complement. It is now well recognized that many pathogenic bacteria, including Borrelia burgdorferi, bind FH/FHL-1 on their cell surface to evade complement-mediated destruction during infection. Recently, it was suggested that B. burgdorferi open reading frame bbA68, known as complement regulator-acquiring surface protein 1 (CRASP-1), encodes the major FH/FHL-1-binding protein of B. burgdorferi. However, because several other proteins have been identified on the surface of B. burgdorferi that also can bind FH/FHL-1, it is presently unclear what role CRASP-1 plays in serum resistance. To examine the contribution of CRASP-1 in serum resistance, we generated a B. burgdorferi mutant that does not express CRASP-1. The B. burgdorferi CRASP-1 mutant, designated B31cF-CRASP-1, was found to be as susceptible to human serum as a wild-type strain of Borrelia garinii 50 known to be sensitive to human serum. To further examine the role of CRASP-1 in serum resistance, we also created a shuttle vector that expresses CRASP-1 from the native B. burgdorferi gene, which was designated pKFSS-1::CRASP-1. When the pKFSS-1::CRASP-1 construct was transformed into the B. burgdorferi B31cF-CRASP-1 mutant, wild-type levels of serum resistance were restored. Additionally, when pKFSS-1::CRASP-1 was transformed into the serum-sensitive B. garinii 50 isolate, human serum resistance was imparted on this strain to a level indistinguishable from wild-type B. burgdorferi. The combined data led us to conclude that CRASP-1 expression is necessary for B. burgdorferi to resist killing by human serum. PMID:16116222

Brooks, Chad S; Vuppala, Santosh R; Jett, Amy M; Alitalo, Antti; Meri, Seppo; Akins, Darrin R



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)



A serum protein involved in aging?  

Microsoft Academic Search

Aging of the brain is characterized, in part, by the appearance of protein anomalies. The proteins deposited within the nervous\\u000a system structures are hardly soluble. This physiological phenomenon turns out to be pathological, quantitatively at least,\\u000a and perhaps even qualitatively, in dementia of the Alzheimer’s type (DAT). One might wonder whether the brain protein anomalies\\u000a are related to a general

K. Z. Ai; K. Vermuyten; P. P. De Deyn; A. Lowenthal; D. Karcher




PubMed Central

1. From a consideration of previous work related to the problem of the influence of diet upon the regeneration of serum protein, a plan of study is described which eliminates the many variables shown to be operating in the studies conducted by these pioneer workers. 2. By the administration of a protein-free diet at a high level of caloric intake to the dog subjected to plasmapheresis during which one-fourth of the blood volume of the animal is withdrawn daily, it is possible to reduce the serum protein concentration to the basal level (3.5 to 4.2 per cent) and to deplete the organism of its reserve stores of this protein within 1 week. The subsequent week has been demonstrated to be an equilibrium period. 3. The dog contains a reserve store of serum protein building material equivalent to about 30 to 40 per cent of the total amount normally present in the circulation. 4. When fed the protein-free diet and when subjected to quantitative plasmapheresis, whereby the basal level of the serum protein concentration is maintained constant, the dog is able in 1 week to regenerate approximately 20 to 30 per cent of the total amount of this blood protein normally present in the plasma. 5. The administration of a diet favorable for promoting the regeneration of serum protein requires approximately 4 to 5 days before a constant and maximal response to the dietary stimulus is obtained. Equilibrium data are yielded during the 2nd week, and these are employed in calculating the potency ratio of the dietary protein.

Melnick, Daniel; Cowgill, George R.; Burack, Ethel



Identification of differentially expressed serum proteins in infectious purpura fulminans.  


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

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



Screening serum hepatocellular carcinoma-associated proteins by SELDI-based protein spectrum analysis  

Microsoft Academic Search

AIM: To find out potential serum hepatocellular carcinoma (HCC)-associated proteins with low molecular weight and low abundance by SELDI-based serum protein spectra analysis, that will have much application in the diagnosis or differentiated diagnosis of HCC, as well as giving a better understanding of the mechanism of hepato-carcinogenesis. METHODS: Total serum samples were collected with informed consent from 81 HCC

Jie-Feng Cui; Yin-Kun Liu; Hai-Jun Zhou; Xiao-Nan Kang; Cheng Huang; Yi-Feng He; Zhao-You Tang; Toshimasa Uemura



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.

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



Serum Protein Profile at Remission Can Accurately Assess Therapeutic Outcomes and Survival for Serous Ovarian Cancer  

PubMed Central

Background Biomarkers play critical roles in early detection, diagnosis and monitoring of therapeutic outcome and recurrence of cancer. Previous biomarker research on ovarian cancer (OC) has mostly focused on the discovery and validation of diagnostic biomarkers. The primary purpose of this study is to identify serum biomarkers for prognosis and therapeutic outcomes of ovarian cancer. Experimental Design Forty serum proteins were analyzed in 70 serum samples from healthy controls (HC) and 101 serum samples from serous OC patients at three different disease phases: post diagnosis (PD), remission (RM) and recurrence (RC). The utility of serum proteins as OC biomarkers was evaluated using a variety of statistical methods including survival analysis. Results Ten serum proteins (PDGF-AB/BB, PDGF-AA, CRP, sFas, CA125, SAA, sTNFRII, sIL-6R, IGFBP6 and MDC) have individually good area-under-the-curve (AUC) values (AUC?=?0.69–0.86) and more than 10 three-marker combinations have excellent AUC values (0.91–0.93) in distinguishing active cancer samples (PD & RC) from HC. The mean serum protein levels for RM samples are usually intermediate between HC and OC patients with active cancer (PD & RC). Most importantly, five proteins (sICAM1, RANTES, sgp130, sTNFR-II and sVCAM1) measured at remission can classify, individually and in combination, serous OC patients into two subsets with significantly different overall survival (best HR?=?17, p<10?3). Conclusion We identified five serum proteins which, when measured at remission, can accurately predict the overall survival of serous OC patients, suggesting that they may be useful for monitoring the therapeutic outcomes for ovarian cancer.

Ghamande, Sharad A.; Bush, Stephen; Ferris, Daron; Zhi, Wenbo; He, Mingfang; Wang, Meiyao; Wang, Xiaoxiao; Miller, Eric; Hopkins, Diane; Macfee, Michael; Guan, Ruili; Tang, Jinhai; She, Jin-Xiong



Serum protein signatures differentiating autoimmune pancreatitis versus pancreatic cancer.  


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

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



Serum Protein Signatures Differentiating Autoimmune Pancreatitis versus Pancreatic Cancer  

PubMed Central

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

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



Detection of serum proteins in the electrophoretic patterns of total proteins of mycoplasma cells.  

PubMed Central

The contamination of mycoplasma cell preparations by serum proteins originating from culture medium was studied. A. laidlawii and M. arthritidis cells were grown in the presence of [14C]-aminoacids, and the cells were washed with 0-9% NaC1 by threefold centrifugation. Total proteins of the washed cells were analysed by SDS gel electrophoresis. Coomassie-stained electrophoretic patterns were compared with autoradiographs of the same gels. The stained electrophoretic pattern of washed A. laidlawii grown without serum was identical with autoradiographs of the same cells grown without or with serum. That of washed A. laidlawii grown with serum differed from the corresponding autoradiography by the presence of extra protein bands I, II, III, and IV with molecular weights of over 160,000, 80,000-87,000, 55,000 and 25,000, respectively. The same extra bands were found in stained electrophoretic patterns of washed: (a) A. laidlawii cells grown without serum and mixed with serum in the stationary phase, (b) M. arthritidis cells, as compared with their autoradiographs, (c) serum precipitate. The bands III and IV may be due to the heavy and light chains of gamma-globulin, the band II might belong to transferrin or to some component of complement. Acidification of serum to pH 5 brought about 100-fold rise of amount of serum precipitate, the number of bands in the electrophoretic pattern of the precipitate being also increased. Stained electrophoretic patterns of cells purified by twofold centrifugation in step sucrose density gradient (1-20-1-27 g./cm.3 for A. laidlawii, and 1-15-1-25 for M. arthritidis) contained no extra bands and matched completely with their autoradiographs. It was concluded that contamination of washed mycoplasma cells by serum proteins is mainly due to co-precipitation of aggregated serum proteins together with cells during centrifugation rather than to adsorption of serum proteins on the cell surface. Images Plate 1

Yaguzhinskaya, O. E.



[Physiocochemical properties of blood serum proteins of coal miners].  


Using disk electrophoresis in the polyacrylamide gel, blood serum proteins were studied in miners working under conditions of the combine (the control group) and drilling-and-blasting (the contact with carbon oxide, nitrogen oxides) driving technique under normal temperature conditions. 26--27 protein fractions characterized by mobility, thermolability under definite conditions of the experiment and the contitative content were obtained. It is shown that the contact with carbon oxide and nitrogen oxides causes changes in the rpoperties of certain proteins (II3, globulins--2 alpha 1, 3 alpha 1, 2 beta, 2 alpha 2, 5 alpha 2, 6 alpha 2, 7 alpha 2) of miners blood serum. Some of these proteins are supposed to participate in the adaptation reactions of the organism. PMID:473394

Nandakova, V N; Zemliakova, L F; Sukhanov, V V; Min'ko, L A



Fractionation of Serum Protein by cm-Cellulose Column.  

National Technical Information Service (NTIS)

Employing CM-cellulose column, serum protein is eluted with pH 5.4 acetate buffer, and the composition of each fraction is identified by the paper electrophoresis of each fraction and the comparison of its chromatogram with that of standard albumin and gl...

Y. Tanaka Y. Iwashita Y. Nakakima T. Kohi S. Noguchi



Fractionation of Serum Proteins with a Quaternary Ammonium Detergent  

Microsoft Academic Search

THE fractionation of serum proteins with the aid of quaternary ammonium compounds has met with difficulties, because the detergent\\/protein ratio must be maintained at a very constant level1. We have shown that this is no longer necessary if `Desogen' (methylphenyl dodecyltrimethyl-ammonium-methosulphate, Geigy S.A., Basle) is used. Separation into at least four components can be performed by the following method.

F. J. Loomeijer



Chemical labelling of active serum thioester proteins for quantification  

PubMed Central

The complement serum proteins C3 and C4 and the protease inhibitor ?-2 macroglobulin are all members of the C3/?-2M thioester protein family, an evolutionarily ancient and conserved family that contains an intrachain thioester bond. The chemistry of the thioester bond is a key to the function of the thioester proteins. All these proteins function by covalently linking to their target by acyl transfer of the protein via the thioester moiety. We show that the signature thioester bond can be targeted with nucleophiles linked to a bioreporter molecule, site-specifically modifying the whole, intact thioester protein. Conditions were optimised to label selectively and efficiently pull-down unprocessed thioester-containing proteins from serum. We demonstrated pull-down of full-length C3, ?-2M and C4 from sera in high salt, using a biotinylated nucleophile and streptavidin-coated resin, confirmed by MALDI-TOF MS identification of the gel bands. The potential for the development of a quantitative method for measuring active C3 in serum was investigated in patient sera pre and post operation. Quantifying active C3 in clinical assays using current methods is difficult. Methods based on antibody detection (e.g. nephelometry) do not distinguish between active C3 and inactive breakdown products. C3-specific haemolytic assays can be used, but these require use of relatively unstable reagents. The current work represents a promising robust, enzyme- and antibody-free chemical method for detecting active thioester proteins in blood, plasma or serum.

Holm, Lotta; Ackland, Gareth L.; Edwards, Mark R.; Breckenridge, Ross A.; Sim, Robert B.; Offer, John



Sensing of proteins in human serum using conjugates of nanoparticles and green fluorescent protein  

Microsoft Academic Search

There is a direct correlation between protein levels and disease states in human serum, which makes it an attractive target for sensors and diagnostics. However, this is challenging because serum features more than 20,000 proteins, with an overall protein content greater than 1 mM. Here we report a sensor based on a hybrid synthetic-biomolecule that uses arrays of green fluorescent

Mrinmoy de; Subinoy Rana; Handan Akpinar; Oscar R. Miranda; Rochelle R. Arvizo; Uwe H. F. Bunz; Vincent M. Rotello



Serum angiopoietin-like protein 3 concentrations in rheumatic diseases.  


Angiopoietin-like protein 3 (Angptl3) is one of the angiogenic cytokines that stimulates endothelial cell adhesion, migration, and neovascularization. No link has been established between Angptl3 and rheumatic diseases such as systemic sclerosis or dermatomyositis (DM). In this study, we determined the serum Angptl3 levels in patients with various rheumatic diseases, and tried to evaluate the possibility that serum levels of Angptl3 can be a useful disease marker. Serum samples were collected from 21 SSc patients, 10 systemic lupus erythematosus (SLE) patients, 21 DM patients, 5 polymyositis (PM) patients and 11 patients with clinically amyopathic DM (CADM) as well as 12 healthy volunteers. Levels of serum Angptl3 were measured with a specific ELISA kit. There was a significant increase of serum Angptl3 levels in patients with SSc or DM (p<0.05). Levels of serum Angptl3 were also slightly higher in patients with ADM, PM or SLE compared with healthy controls, but not statistically significant. Myoglobin levels were significantly higher in DM patients with increased serum Angptl3 levels than those with normal levels (p<0.05). In addition, among patients with SSc, the prevalence of cutaneous ulcers was significantly greater in patients with elevated Angptl3 levels than those with normal levels (p<0.05). Serum Angptl3 levels may be associated with the pathogenesis of muscle involvement in DM patients and microangiopathy in SSc patients. Clarifying the role of Angptl3 in each rheumatic disease may lead to further understanding of the pathogenesis and new therapeutic approaches. PMID:22652544

Hayashi, Yasuyo; Jinnin, Masatoshi; Makino, Takamitsu; Kajihara, Ikko; Makino, Katsunari; Honda, Noritoshi; Nakayama, Wakana; Inoue, Kuniko; Fukushima, Satoshi; Ihn, Hironobu



Identification of protein carbonyls in serum of the fetal and neonatal pig  

Microsoft Academic Search

Oxidation of serum proteins leads to non-reversible carbonyl formation which alters their function and is associated with stress-related disease processes. The primary objective of this study was to quantify and identify oxidized serum proteins in fetal and newborn piglets. Protein carbonyls were converted to hydrazones with dinitrophenyl hydrazine and quantified spectrophotometrically. For identification, serum protein carbonyls were derivatized with biotin

Thomas J. Caperna; Amy E. Shannon; Le Ann Blomberg; Wesley M. Garrett; Timothy G. Ramsay



Interactions of lecithinized superoxide dismutase with serum proteins and cells.  


Superoxide dismutase covalently bound to four lecithin molecules (PC-SOD) is known to be retained in circulating blood for a prolonged period and has a high affinity for cells, resulting in beneficial therapeutic effects in animal disease models. In this study, we evaluated the interaction of PC-SOD with biological components, such as serum proteins and cells, to clarify the mechanism underlying the improved pharmacokinetics of SOD induced by lecithin chemical modification (lecithinization). PC-SOD was distributed in the plasma but not in blood cells after being added to the blood. PC-SOD formed a complex with serum protein(s) such as albumin, whereas unmodified SOD did not. The cellular content of PC-SOD was markedly higher than that of unmodified SOD, and was distributed in lysosomes. The pathway associated with the cellular uptake was found to involve clathrin-/caveolae-independent and cholesterol-sensitive endocytosis. Overall, our data indicated that the increased hydrophobicity of lecithinized SOD enhanced its association to both serum protein(s) and plasma membrane microdomains. The former inhibited SOD excretion and promoted long-term retention in circulating blood, whereas the latter enhanced internalization into cells via endocytosis. © 2014 Wiley Periodicals, Inc. and the American Pharmacists Association J Pharm Sci 103:1987-1994, 2014. PMID:24867597

Ishihara, Tsutomu; Nara, Shunsuke; Mizushima, Tohru



Identification of serum proteins bound to industrial nanomaterials.  


Nanoparticles (NPs) are decorated with proteins and other biomolecules when they get into contact with biological systems. The presence of proteins in cell culture medium can therefore have effects on the biological outcome in cell-based tests. In this study, the manufactured nanomaterials silicon dioxide (SiO(2)), titanium dioxide (TiO(2)), iron-III-oxide (Fe(2)O(3)), and carbon black (CB) were used to study their interaction with single proteins from bovine and human plasma (albumin, fibrinogen and IgG) as well as with complete human serum. The protein binding capacity of the material was investigated and 1D gel electrophoresis was used to separate the bound proteins and to identify the bands by matrix-assisted laser desorption/ionisation-time-of-flight (MALDI-TOF) mass spectrometry. We found that the NP surface chemistry had a great impact on the amount of bound protein with distinct ligands for each of the tested particles. The hydrophobic CB NPs bound much more protein than the hydrophilic metal oxide NPs. Among the single proteins investigated, fibrinogen showed the strongest affinity for SiO(2), TiO(2) and CB NPs. The identified proteins from human serum adsorbed to these NPs were very different. Only apolipoprotein A1 was found to be adsorbed to all NPs. These studies will help to explain the different degree of biological responses observed after in vitro exposure of cells in the absence or presence of serum and might also support the interpretation of in vivo experiments were NPs come directly into contact with blood plasma. PMID:22001751

Ruh, Hermelindis; Kühl, Boris; Brenner-Weiss, Gerald; Hopf, Carsten; Diabaté, Silvia; Weiss, Carsten



Distribution of selenium-containing proteins in human serum  

Microsoft Academic Search

Selenium-containing proteins in human serum of four volunteers in Beijing were separated and purified by preparative sodium\\u000a dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Selenium contents in the proteins were quantified by high-performance\\u000a liquid chromatography (HPLC) coupled with a fluorescence detector (FLD) after pretreatment with a microwave digestion system\\u000a and derivatization with 2,3-diaminonaphthalene (DAN). Five selenium-containing proteins with apparent molecular weights (MWs)

Yuxi Gao; Yingbin Liu; Guilong Deng; Zijian Wang



Heparin chromatography to deplete high-abundance proteins for serum proteomics  

Microsoft Academic Search

BackgroundSerum is a very informative sample for disease diagnosis. However, a few of the high-abundance proteins existing in serum make the identification of disease-specific serum biomarkers extremely challenging using currently available technologies. A highly promising first step for most analytical approaches of serum is to deplete as many of the high-abundance proteins as possible.

Ting Lei; Qing-Yu He; Yi-Li Wang; Lu-Sheng Si; Jen-Fu Chiu



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


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



Serum proteins and paraproteins in women with silicone implants and connective tissue disease: a case–control study  

Microsoft Academic Search

Prior studies have suggested abnormalities of serum proteins, including paraproteins, in women with silicone implants but did not control for the presence of connective-tissue disease (CTD). This retrospective case–control study, performed in tertiary-care academic centers, assessed possible alterations of serum proteins, including paraproteins, in such a population. Seventy-four women with silicone implants who subsequently developed CTD, and 74 age-matched and

Gyorgy Csako; Rene Costello; Ejaz A Shamim; Terrance P O'Hanlon; Anthony Tran; Daniel J Clauw; H James Williams; Frederick W Miller



Etude Systematique des Proteines Plasmatiques et Seriques du Porc (Systematic Study of Plasma and Serum Proteins in the Pig).  

National Technical Information Service (NTIS)

The serum and plasma proteins of a normal pig were studied to discover whether the qualitative and quantitative changes in these proteins can make a significant contribution to the establishment of a biological dosimetry for irradiated pigs. The serum and...

F. Daburon C. Hatchikan J. P. Schmidt P. Nizza



Elevated expression of RGS19 impairs the responsiveness of stress-activated protein kinases to serum.  


Regulators of G protein signaling (RGS proteins) serve as GTPase activating proteins for the signal transducing G? subunits. RGS19, also known as G?-interacting protein (GAIP), has been shown to subserve other functions such as the regulation of macroautophagy and growth factor signaling. We have recently demonstrated that the expression of RGS19 in human embryonic kidney (HEK) 293 cells resulted in the disruption of serum-induced mitogenic response along the classical Ras/Raf/MEK/ERK pathway. Here, we further examined the effect of RGS19 expression on the stress-activated protein kinases (SAPKs). Both c-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase (MAPK) became non-responsive to serum in 293/RGS19 cells, yet the two SAPKs responded to UV irradiation or osmotic stress induced by sorbitol. Kinases upstream of JNK and p38 MAPK, including MKK3/6, MKK4, and MLK3, also failed to respond to serum stimulation in 293/RGS19 cells. Serum-induced activation of the small GTPases Rac1 and Cdc42 was similarly suppressed in these cells. Our results indicate that elevated expression of RGS19 can severely disrupt the regulation of MAPKs by small GTPases. PMID:22045062

Ip, Angel K C; Tso, Prudence H; Lee, Maggie M K; Wong, Yung H



Hydroactive dressings and serum proteins: an in vitro study.  


An in vitro approach was used to obtain information about the compatibility of hydroactive dressing materials with the serum proteins which are thought to be relevant to wound healing. Artificial wound fluid was incubated with different hydroactive dressings (Cutinova hydro, Varihesive E, Comfeel Ulcer Dressing and Allevyn), and concentrations of total protein, albumin, immunoglobulin and growth factors were measured after one day of incubation. Cutinova hydro and Allevyn absorbed considerable amounts of fluid. Fluid uptake was lower for the hydrocolloid dressings. An unexpected finding was that Cutinova hydro showed an approximately two-fold increase over control values in the concentration of all proteins tested, indicating a selective absorption of water by this dressing. For the other dressings tested, zero or very low absorption of proteins was found, indicating a basically satisfactory protein compatibility. PMID:8697137

Achterberg, V; Meyer-Ingold, W



Search for serum protein-binding disaccharides and disaccharide-binding serum proteins by affinity capillary electrophoresis.  


The potential use of affinity capillary electrophoresis in a microscale search for mutually interacting substances in biological fluid is demonstrated. Some disaccharides, especially gentiobiose (Gen), derivatized with 1-phenyl-3-methyl-5-pyrazolone, caused peak retardation when electrophoresed in a neutral running buffer, containing human serum. Gen, the most significantly retarded disaccharide, was converted to its negatively charged bis-mercaptoethanesulfonate derivative (MerESGen), and a serum sample was analyzed in a neutral buffer containing the derivatized disaccharide. Two peaks, belonging to the beta-globulin fraction, were found to be remarkably retarded in the buffer containing MerES-Gen in a concentration-dependent way. These findings prove an interaction between disaccharides and serum proteins. PMID:15004849

Taga, Atsushi; Yamamoto, Yuka; Maruyama, Rie; Honda, Susumu



[Concentration of serum proteins in children with down syndrome].  


In the serum of 41 children with trisomy 21 (Down syndrome) the concentration was determined of 10 proteins by the radial immunodiffusion method. It was found that the concentration of prealbumin and alpha 2-macroglobulin was lower than in healthy children. The concentration of acid alpha 1-glycoprotein, alpha 1-antitrypsin, haptoglobin, C4 complement component, and hemopexin was higher than in healthy children. The concentration of ceruloplasmin, C3 complement component, and transferrin was similar as in healthy children. PMID:1295255

Bartelik, S



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.



Study of arsenic-protein binding in serum of patients on continuous ambulatory peritoneal dialysis  

Microsoft Academic Search

Arsenic (As) bound to serum proteins in patients on continuous ambulatory peritoneal dialysis (CAPD) was studied. A prior experiment by ultrafiltration showed that 5.57% of total As was bound to serum proteins for 14 CAPD patients. Further identification of the As species and protein molecules in serum of three CAPD patients with high As concentrations was carried out by combining

Xinrong Zhang; Rita Cornelis; Jurgen De Kimpe; Louis Mees; Norbert Lameire


Specific Changes of Serum Proteins in Parkinson's Disease Patients  

PubMed Central

The aim of this study is to identify and validate protein change in the serum from PD patients. We used serum samples from 21 PD patients and 20 age-matched normal people as control to conduct a comparative proteomic study. We performed 2-DE and analyzed the differentially expressed protein spots by LC-MS/MS. In PD group 13 spots were shown to be differentially expressed compared to control group. They were identified as 6 proteins. Among these, 3 proteins were confirmed by Western blot analysis. It showed that the frequency of fibrinogen ?-chain (FGG) appeared 70% in PD, which could not be detected in control group. The protein of inter-alpha-trypsin inhibitor heavy chain H4 (ITI-H4) was found to exist two forms in serum. The full size (120 kDa) of the protein was increased and the fragmented ITI-H4 (35 kDa) was decreased in PD group. The ratio of full size ITI-H4 to fragmented ITI-H4 in PD patients was 3.85±0.29-fold higher than in control group. Furthermore, fragmented Apo A-IV (?26 kDa) was mainly detected in control group, while it was rare to be found in PD group. Above findings might be useful for diagnosis of PD. When the expressions of FGG and 120 kDa ITI-H4 are increase, as well as ?26 kDa Apo A-IV disappear would provide strong evidence for PD.

Lu, Wenwen; Wan, Xinhua; Liu, Bin; Rong, Xianfang; Zhu, Lei; Li, Pingping; Li, Jiang; Wang, Ling; Cui, Liying; Wang, Xiaoliang



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


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



Abnormal serum concentrations of proteins in Parkinson's disease.  


Blood serum was used to identify protein biomarkers for diagnosis of Parkinson's disease (PD) using analytically validated quantitative 2D-gel electrophoresis, and single variable and multivariate statistics. Using banked samples from a first medical center, we identified 57 specific protein spot biomarkers with disease-specific abnormal levels in serum of patients with PD, Alzheimer's disease, amyotrophic lateral sclerosis and similar neurodegenerative conditions (337 samples), when compared to age-matched normal controls (132 samples). To further assess their clinical usefulness in Parkinson's disease, we obtained prospective newly drawn blood serum samples from a second (56 PD, 30 controls) and third (6 PD, 48 controls) medical center. The protein concentrations of the 57 biomarkers were assessed by 2D-gel electrophoresis. Stepwise linear discriminant analysis selected a combination of 21 of the 57 as optimal to distinguish PD patients from controls. When applied to the samples from the second site, the 21 proteins had sensitivity of 93.3% (52 of 56 PD correctly classified), specificity of 92.9% (28 of 30 controls correctly classified); 15 of 15 patients with mild, 28 of 30 with moderate to severe symptoms, and all of the 6 PD patients from the third site were correctly classified. Eleven of the 21 proteins showed statistically significant abnormal concentrations in patients with mild symptoms, and 14 in patients with moderate-severe symptoms. The protein identities reflect the heterogeneity of Parkinson's disease, and thus may provide the capability of monitoring the blood for a diverse range of PD pathophysiological mechanisms: cellular degeneration, oxidative stress, inflammation, and transport. PMID:19723509

Goldknopf, Ira L; Bryson, Jennifer K; Strelets, Irina; Quintero, Silvia; Sheta, Essam A; Mosqueda, Miguel; Park, Helen R; Appel, Stanley H; Shill, Holly; Sabbagh, Marwan; Chase, Bruce; Kaldjian, Eric; Markopoulou, Katerina



Interactions of apomorphine with serum and tissue proteins  

SciTech Connect

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

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



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.

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



Studies on the Serum Proteins IV. The Dye-Binding of Purified Serum Proteins Separated by Continuous-Flow Electrophoresis  

Microsoft Academic Search

HE FRACTIONATION OF SERUM PROTEINS by paper electrophoresis, first described in 1949 (1), has had ever-widening clinical application dur- ing the past decade. Discrepancies between quantitative analyses by paper electrophoresis and by the older, moving-boundary technics stimulated the development of conversion factors to render compara- ble the results obtained by the 2 methods (2-8). These conversion fac- tors have been

F. William Sunderman


Protein Microarrays for Quantitative Detection of PAI-1 in Serum  

PubMed Central

Objective Plasminogen activator inhibitor-1 (PAI-1), one crucial component of the plasminogen activator system, is a major player in the pathogenesis of many vascular diseases as well as in cancer. High levels of PAI-1 in breast cancer tissue are associated with poor prognosis. The aim of this study is to evaluate rigorously the potential of serum PAI-1 concentration functioning as a general screening test in diagnostic or prognostic assays. Methods A protein-microarray-based sandwich fluorescence immunoassay (FIA) was developed to detect PAI-1 in serum. Several conditions of this microarray-based FIA were optimized to establish an efficacious method. Serum specimens of 84 healthy women and 285 women with breast cancer were analyzed using the optimized FIA microarray. Results The median serum PAI-1 level of breast cancer patients was higher than that of healthy women (109.7 ng/ml vs. 63.4 ng/ml). Analysis of covariance revealed that PAI-1 levels of the two groups were significantly different (P<0.001) when controlling for an age effect on PAI-1 levels. However, PAI-1 values in TNM stage I?IV patients respectively were not significantly different from each other. Conclusion This microarray-based sandwich FIA holds potential for quantitative analysis of tumor markers such as PAI-1.

Ma, Xu



Maternal serum insulin-like growth factor-binding protein-3 (IGFBP-3) at 11–13 weeks in preeclampsia  

Microsoft Academic Search

The objective of this study was to determine if the maternal serum concentration of insulin-like growth factor-binding protein-3 (IGFBP-3) at 11–13 week's gestation is altered in pregnancies that subsequently develop preeclampsia (PE). Maternal serum concentration of IGFBP-3, pregnancy-associated plasma protein-A (PAPP-A) and uterine artery pulsatility index (PI) were measured in 60 cases that developed PE, including 20 that developed early-PE

S Sifakis; R Akolekar; D Kappou; N Mantas; K H Nicolaides



p53 protein is absent from the serum of patients with lung cancer.  

PubMed Central

p53 protein, which accumulates intracellularly in over half of all human tumours, has also been reported to be present in the sera of patients with various malignancies, including lung cancer. Using a quantitative immunoassay, we measured p53 protein concentrations in 216 sera from 114 lung cancer patients of whom 75 provided matched lung tumour tissues, which were also assayed for p53 protein. p53 protein levels above the detection limit of 0.04 ng ml-1 were detected in only two sera from lung cancer patients (0.14 ng ml-1 and 0.27 ng ml-1), but not in any of 13 sera from non-malignant lung disease patients or in 100 sera from normal non-diseased individuals. The presence of these apparent traces of serum p53 protein concentrations could not be related either to the p53 protein expression status of the primary lung tumours or to the tumour stage, grade or histological type. By pretreating these two sera with anti-p53 antibody linked to solid phase, and by the addition of mouse serum to neutralise possible heterophilic antibodies, the signals arising from these sera were shown to be non-specific and possibly caused by heterophilic antibodies. We conclude that our data do not support previous reports of p53 protein in the sera of lung cancer patients. Since immunoassays are subject to numerous sources of interference in serum, including heterophilic antibodies, we suggest that the results of p53 protein analysis of serum specimens should be interpreted with caution.

Levesque, M. A.; D'Costa, M.; Diamandis, E. P.



Oxidative modification of serum proteins in multiple sclerosis.  


Multiple sclerosis (MS) has been demonstrated to involve oxidative stress and augmented glycoxidation. In this study, several markers of protein oxidative damage and glycoxidation have been compared in 14 relapsing remittent in MS (RRMS) patients without immunomodifying treatment, 10 patients in clinical relapse, and clinically stable patient groups treated with interferon ? 1a (18) , ? 1b (19) and glatiramer acetate (GA; 6) in relation to healthy subjects (12). The glycophore content was increased in RRSM patients without treatment and in patients treated with GA. The level of advanced protein oxidation products (AOPP) was increased in RRSM patients without treatment and in patients with clinical relapse. The level of protein carbonyls was elevated in RRSM patients without treatment and in patients treated with interferon ? 1b. The levels of dityrosine level and N'-formylkynureine were elevated in RRSM patients without treatment while serum protein thiol groups were decreased in RRSM patients in clinical relapse as well as RRMS patients treated with interferon ? 1a. Several markers of protein modification showed correlation with the C-reactive protein level and white blood cell count, suggesting that oxidative protein modifications are linked to the inflammatory processes in MS. Results of this study confirm the occurrence of protein oxidative and glycoxidative damage in MS and show that spectrophotometric and fluorimetric markers of this damage, especially the AOPP level, may be useful in monitoring oxidative stress in the course of therapy of MS. PMID:24036284

Sadowska-Bartosz, Izabela; Adamczyk-Sowa, Monika; Galiniak, Sabina; Mucha, Sebastian; Pierzchala, Krystyna; Bartosz, Grzegorz



Photo selective protein immobilization using bovine serum albumin  

NASA Astrophysics Data System (ADS)

A simple and selective technique which immobilizes protein onto a solid substrate by using UV illumination has been developed. In protein immobilization, a Bovine serum albumin (BSA) performed bifunctional role as a cross-linker between substrate and proteins and as a blocker inhibiting a nonspecific protein adsorption. A new photo-induced protein immobilization process has been investigated at each step by fluorescence microscopy, ellipsometry, and Fourier transform infrared (FT-IR) spectroscopy. A UV photomask has been used to induce selective protein immobilization on target regions of the surface of the SiO2 substrates under UV illumination with negligible nonspecific binding. The UV illumination also showed improved photostability than the conventional methods which employed bifunctional photo-crosslinker molecules of photo-reactive diazirine. This new UV illumination-based photo-addressable protein immobilization provides a new approach for developing novel protein microarrays for multiplexed sensing as well as other types of bio-immobilization in biomedical devices and biotechnologies.

Kim, Wan-Joong; Kim, Ansoon; Huh, Chul; Park, Chan Woo; Ah, Chil Seong; Kim, Bong Kyu; Yang, Jong-Heon; Chung, Kwang Hyo; Choi, Yo Han; Hong, Jongcheol; Sung, Gun Yong



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.

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



Serum and Antibodies of Glaucoma Patients Lead to Changes in the Proteome, Especially Cell Regulatory Proteins, in Retinal Cells  

PubMed Central

Purpose Previous studies show significantly specifically changed autoantibody reactions against retinal antigens in the serum of glaucoma and ocular hypertension (OHT) patients in comparison to healthy people. As pathogenesis of glaucoma still is unknown the aim of this study was to analyze if the serum and antibodies of glaucoma patients interact with neuroretinal cells. Methods R28 cells were incubated with serum of patients suffering from primary open angle glaucoma (POAG), normal tension glaucoma (NTG) or OHT, POAG serum after antibody removal and serum from healthy people for 48 h under a normal or an elevated pressure of 15000 Pa (112 mmHg). RGC5 cells were additionally incubated with POAG antibodies under a normal pressure. Protein profiles of the R28 cells were measured with Seldi-Tof-MS, protein identification was performed with Maldi-TofTof-MS. Protein analysis of the RGC5 cells was performed with ESI-Orbitrap MS. Statistical analysis including multivariate statistics, variance component analysis as well as calculating Mahalanobis distances was performed. Results Highly significant changes of the complex protein profiles after incubation with glaucoma and OHT serum in comparison to healthy serum were detected, showing specific changes in the cells (e.g. Protein at 9192 Da (p<0.001)). The variance component analysis showed an effect of the serum of 59% on the cells. The pressure had an effect of 11% on the cells. Antibody removal led to significantly changed cell reactions (p<0.03). Furthermore, the incubation with POAG serum and its antibodies led to pro-apoptotic changes of proteins in the cells. Conclusions These studies show that the serum and the antibodies of glaucoma patients significantly change protein expressions involved in cell regulatory processes in neuroretinal cells. These could lead to a higher vulnerability of retinal cells towards stress factors such as an elevated IOP and eventually could lead to an increased apoptosis of the cells as in glaucoma.

Bell, Katharina; Funke, Sebastian; Pfeiffer, Norbert; Grus, Franz H.



Blood Antigen, Serum Protein, and Milk Protein Gene Frequencies and Genetic Interrelationships in Holstein Cattle1  

Microsoft Academic Search

Gene frequencies at ten blood group loci, one serum protein locus, and four milk protein loci were determined for the Holstein-Friesian breed in the United States. The sample consisted of 8630 cows in 51 herds from 10 states. Because of the close linkage among casein subloci and the concomitant rarity of crossover- recombinant groups, casein gene com- bination or haplotype

H. C. Hines; G. F. W. Haenlein; J. P. Zikakis; H. C. Dickey



Glomerular expression and elevated serum Bcl-2 and Fas proteins in lupus nephritis: preliminary findings  

PubMed Central

Programmed cell death (apoptosis) is involved in glomerular injuries leading to glomerulonephritis. Bcl-2 and Fas are proteins that promote cell survival and death, respectively. This study tests the hypothesis that lupus nephritis is associated with alterations of Bcl-2 and Fas protein expression. Thirty-six patients with lupus nephritis and 10 controls (normal individuals) were included in this study. Bcl-2 and Fas positive cells were examined in kidney biopsies by immunohistochemistry. Bcl-2 and Fas serum levels were evaluated by enzyme-linked immunosorbent assay (ELISA). In the glomeruli of normal kidneys, Bcl-2 and Fas proteins were completely absent. In lupus nephritis patients, glomerular expression of Bcl-2 and Fas was seen in mesangial cells (1·3 ± 0·1 and 2·0 ± 0·1 for Bcl-2 and Fas, respectively). Similarly, a statistically significantly higher Bcl-2 (217·1 ± 85·9) and Fas (767·9 ± 271) serum levels were found in lupus patients compared to controls (148·6 ± 87, 550·3 ± 91 for Bcl-2 and Fas, P < 0·05). A direct correlation between serum Bcl-2 and Fas and chronicity index was also found. Compared to normal controls, lupus nephritis is associated with glomerular expression and elevated serum levels of Bcl-2 and Fas proteins. These findings suggest possible roles for Bcl-2 and Fas in glomerular injury during evolution of lupus nephritis. The diagnostic, prognostic and therapeutic ramifications of our findings are open to further investigation.

Fathi, N A; Hussein, M R; Hassan, H I; Mosad, E; Galal, H; Afifi, N A



A comparative study of milk serum proteins in camel ( Camelus dromedarius) and bovine colostrum  

Microsoft Academic Search

Camel (Camelus dromedarius) whey proteins were detected and compared to bovine whey proteins using size exclusion chromatography columns on HPLC. Camel whey proteins such as serum albumin and ?-lactalbumin appear to possess molecular weights similar to the respective bovine whey proteins. Camel whey lacks ?-lactoglobulin and consists of large amount of serum albumin, compared to bovine whey. Camel colostrum is

U Merin; S Bernstein; A Bloch-Damti; R Yagil; C van Creveld; P Lindner; N Gollop



Population genetic studies in the Balkans. I. Serum proteins.  


Within a study of the genetics of Southeastern European populations seven serum protein polymorphisms (AMY2, BF, C3, CP, GC, HPA, TF) were examined in three samples of Aromuns (Albania: the village of Andon Poci, province Gjirocaster, Republic of Macedonia: Stip region, Romania: the village Kogalniceanu, province Dobruja) and four reference samples (Albanians: Tirana, Romanians: Constanta and Ploiesti as well as Greeks (Northeastern Greece)). The Aromun samples from Albania and Romania form one separate cluster and the reference samples together with the Aromuns from Macedonia (Stip region) form a second one. PMID:11591047

Scheil, H G; Scheffrahn, W; Schmidt, H D; Huckenbeck, W; Efremovska, L; Xirotiris, N



Removal of background interference in nephelometric determination of serum proteins.  


By precipitation of beta-lipoprotein by a dextran sulphate and calcium chloride mixture the background light scattering in continuous flow nephelometric systems can be reduced to such a level that it can be neglected in practice. No loss of IgG, IgA, IgM, orosomucoid or transferrin on precipitation could be demonstrated. The supernatant could not be used for immunoelectrophoresis or radial immunodiffusion in analysis of these proteins since significantly lower values were obtained for some of them than in native serum. PMID:912903

Kallner, A



Serum proteins with NAD + glycohydrolase activity and anti-CD38 reactivity – elevated levels in serum of tumour patients  

Microsoft Academic Search

NAD+ glycohydrolase activities in serum samples from cancer patients were two- to three-fold higher than the activities in samples from healthy controls. SDS-PAGE analysis of serum samples followed by Western blotting revealed the presence of two proteins of ~45 and ~21 kDa that were immunoreactive with human CD38-specific monoclonal antibodies T16, HIT2 and OKT10. These proteins appeared to be more

Ça?atay Korkut; Leman Yalç?ntepe; Ne?e Kiremit-Korkut; Semire Uzun-Alt?nöz; Saliha ??sever; Füsun Gümü?el; Demir Tiryaki; Engin Bermek



Inflammation and dietary protein intake exert competing effects on serum albumin and creatinine in hemodialysis patients  

Microsoft Academic Search

Inflammation and dietary protein intake exert competing effects on serum albumin and creatinine in hemodialysis patients.BackgroundCross-sectional studies have shown an inverse correlation between serum C-reactive protein (CRP) and serum albumin concentration in hemodialysis patients. The net effects of inflammation and dietary protein intake on nutritional markers over time are unknown.MethodsTo explore the effects of CRP and normalized protein catabolic rate

George A Kaysen; Glenn M Chertow; Rohini Adhikarla; Belinda Young; Claudio Ronco; Nathan W Levin



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



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

SciTech Connect

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

Klein, N.W.



Serum chemistry alterations, including creatine kinase isoenzymes, in furazolidone toxicosis of ducklings: preliminary findings.  


Furazolidone induces a cardiotoxicosis when fed in toxic concentrations to newly hatched ducklings. This preliminary experiment was designed to determine if creatine kinase (CK) isoenzymic activities or other serum analytes would be useful as indicators of these cardiac alterations. Sera from 12 ducklings (six fed a control ration and six fed the control ration with 700 mg furazolidone added per kg of feed [700 ppm] for 28 days) were analyzed for CK isoenzymic activities, electrolytes, nitrogenous metabolites, hepatic enzymic activities, bilirubin, and glucose. Statistically significant differences between control and treated groups were detected for creatine kinase MB (CK-MB, cardiac muscle origin) isoenzymic activity and bilirubin, potassium, calcium, and total carbon dioxide concentrations. Differences other than CK-MB isoenzymic activity were generally explained by factors related to the toxicosis or sample handling. These findings suggest that CK-MB isoenzymic activity may be useful to detect and monitor the progress of cardiac injury in furazolidone toxicosis, thereby increasing the usefulness of this model of dilated cardiomyopathy. Our findings, analyzed on the Kodak Ektachem 700 Dry Chemistry Analyzer, are compared with serum chemistry values reported in the literature. PMID:1785996

Webb, D M; DeNicola, D B; Van Vleet, J F



The Discrimination between Magnesium and Manganese by Serum Proteins  

PubMed Central

Magnesium and manganese have proved physically and functionally interchangeable in many isolated biological systems investigated in vitro. This lack of discrimination contrasts sharply with the high biological specificity exhibited by intact mammals under a large variety of conditions. The dichotomy between intact animals and their isolated systems might be due at least partially to presence vs. absence of an intact circulation. Hence the capability of mammalian plasma to discriminate between the alkaline earth and the transition metal was investigated by means of equilibrium dialysis, exchange, ultrafiltration, ultracentrifugation, and zone electrophoresis. The states of the respective elements are thus contrasted as follows: (a) Magnesium is partially bound, manganese totally. (b) Magnesium is nonselectively bound by serum proteins, manganese selectively by a ?1-globulin. (c) Under conditions approaching physiological, the two metals do not interchange. This is interpreted as indicating that the plasma proteins contribute to biological specificity by discriminating between a trace metal and a macronutrient.

Foradori, A. C.; Bertinchamps, Albert; Gulibon, Jane M.; Cotzias, George C.



PIXE analysis of blood serum proteins, separated by gel filtration  

NASA Astrophysics Data System (ADS)

Gel filtration of blood serum was carried out with Sephadex G-200 gel and tris buffer at pH 7.8. The amount of protein in each of about 100 collected fractions was monitored using UV absorption. The fractions were concentrated and deposited onto Kimfol backing foils. From the PIXE results distributions of S, Fe, Cu and Zn were obtained in relation to the protein peaks, while elements like K, Ca and Br appeared at the position corresponding to small molecular sizes. Quantification, taking into account the intermediate thicknesses of the samples, gave reasonable values. By adjusting the pH of the tris buffer with acetate instead of HCl, an increase in sensitivity was achieved, as the Cl is an important source of electron bremsstrahlung. By low temperature ashing still better detection limits can be reached, but at the expense of the loss of volatile elements.

Pallon, Jan; Pakarinen, Pirjo; Akselsson, Roland



Characterization of human serum mannan-binding protein promoter.  


Serum mannan binding protein (MBP), a mannose/N-acetylglusosamine-specific lectin, is important in innate immunity. In order to elucidate the mechanism underlying the wide intra- and interracial variety in the MBP serum level, we have studied the transcriptional regulation of human MBP. Rapid amplification of cDNA ends (5' RACE) analysis of Hep G2 RNA indicated the presence of a novel exon, designated as "exon 0," upstream of previously identified exon 1 [Taylor, M.E. et al. (1989) Biochem. J. 262, 763-771]. Two MBP mRNAs with different sized 5'-noncoding regions were detected: the longer transcript starts at exon 0 and the shorter one at exon 1. Promoter analysis involving a luciferase assay vector revealed that the transcript starting from exon 1 predominates over that starting from exon 0. In addition, a hepatocyto-specific nuclear factor, (HNF)-3, which is known to control the expression of hepatocyto-specific genes, up-regulates the transcription of human MBP from exon 1, while a glucocorticoid, which is known to up-regulate acute phase proteins, markedly suppresses MBP transcription. Recently, polymorphisms were found to occur in the promoter region at two positions [Madsen, H.O. et al. (1995) J. Immunol. 155, 3013-3020]. Functional promoter analysis indicated that three haplotype variants as to these positions, HY, LY, and LX, exhibit high, medium and low promoter activity, respectively, in accordance with the results of a previous population study. PMID:10578050

Naito, H; Ikeda, A; Hasegawa, K; Oka, S; Uemura, K; Kawasaki, N; Kawasaki, T



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.

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



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.

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



Serum amyloid-A protein and serum rheumatoid factor as serological surrogate markers for smoking risk factor in Saudi population.  


Tobacco smoking represents major national and international health hazard that interferes with wide range of physiological functions and biomarkers. In the current study we have investigated the influence of tobacco smoking on some biological markers such as serum amyloid protein-A, rheumatoid factor, serum glucose level and lipid profile in Saudi population. The fore mentioned parameters were investigated in a total of 275 cases in 3 different age categories (less than 20 years old, 20-40 years old and older than 40 years old). Long term survey was adopted in all cases; yet, lightly smoking and heavily smoking groups were compared to never smoking healthy population. Results obtained showed significant increase in serum amyloid protein-A and rheumatoid factor in correlation to the degree of smoking nonetheless in the age category older than 40 years old. Serum glucose, triglyceride, and total cholesterol was not affected by smoking in all studied age categories; however serum LDL-cholesterol was elevated and serum HDL-cholesterol was depressed in correlation to the degree of smoking in all age categories. In conclusion, tobacco smoking represents major cardiovascular risk factor in Saudi population in all age categories and serum amyloid protein-A and rheumatoid factor might be used as a serological surrogate marker for such risk. PMID:23455190

Al-Sieni, Abdulbasit Ibraheem; Al-Alawy, Adel Ibraheem; Al-Shehri, Zafer Saad; Al-Abbasi, Fahad Ahmed



Deep depletion of abundant serum proteins reveals low-abundant proteins as potential biomarkers for human ovarian cancer.  


Epithelial ovarian cancer (EOC) ranks fifth as a cause of cancer deaths in women. Current diagnostic and monitoring markers have limited reliability for the detection of disease. We have tested the possibility of identifying candidate biomarkers present at low nanogram to picogram levels after removing both the 12 most abundant and 77 moderately abundant proteins from serum samples of EOC patients using antibody affinity columns. We showed that this approach allows the identification of proteins that are expressed at nanogram per liter levels in the serum. Using ICAT/MS/MS analysis, we identified 51 proteins that are differentially expressed by at least twofold. These proteins include leucine-rich alpha-2-glycoprotein, matrix metalloproteinase-9 (MMP-9), inter-alpha-trypsin inhibitor heavy chain H1, insulin-like growth factor-binding protein 6, insulin-like growth factor-binding protein 3, isoform 1 of epidermal growth factor receptor, angiopoietin-like protein 3 (ANGPTL3) and phosphatidylcholine-sterol acyltransferase. We confirmed the differential expression of MMP9 and ANGPTL3 in normal and ovarian cancer sera by ELISA assays. Further robust clinical evaluation of the candidate markers identified is necessary. PMID:20559449

Lin, Biaoyang; White, James T; Wu, Jian; Lele, Shashikant; Old, Lloyd J; Hood, Leroy; Odunsi, Kunle



Serum protein profiles predict coronary artery disease in symptomatic patients referred for coronary angiography  

PubMed Central

Background More than a million diagnostic cardiac catheterizations are performed annually in the US for evaluation of coronary artery anatomy and the presence of atherosclerosis. Nearly half of these patients have no significant coronary lesions or do not require mechanical or surgical revascularization. Consequently, the ability to rule out clinically significant coronary artery disease (CAD) using low cost, low risk tests of serum biomarkers in even a small percentage of patients with normal coronary arteries could be highly beneficial. Methods Serum from 359 symptomatic subjects referred for catheterization was interrogated for proteins involved in atherogenesis, atherosclerosis, and plaque vulnerability. Coronary angiography classified 150 patients without flow-limiting CAD who did not require percutaneous intervention (PCI) while 209 required coronary revascularization (stents, angioplasty, or coronary artery bypass graft surgery). Continuous variables were compared across the two patient groups for each analyte including calculation of false discovery rate (FDR ? 1%) and Q value (P value for statistical significance adjusted to ? 0.01). Results Significant differences were detected in circulating proteins from patients requiring revascularization including increased apolipoprotein B100 (APO-B100), C-reactive protein (CRP), fibrinogen, vascular cell adhesion molecule 1 (VCAM-1), myeloperoxidase (MPO), resistin, osteopontin, interleukin (IL)-1?, IL-6, IL-10 and N-terminal fragment protein precursor brain natriuretic peptide (NT-pBNP) and decreased apolipoprotein A1 (APO-A1). Biomarker classification signatures comprising up to 5 analytes were identified using a tunable scoring function trained against 239 samples and validated with 120 additional samples. A total of 14 overlapping signatures classified patients without significant coronary disease (38% to 59% specificity) while maintaining 95% sensitivity for patients requiring revascularization. Osteopontin (14 times) and resistin (10 times) were most frequently represented among these diagnostic signatures. The most efficacious protein signature in validation studies comprised osteopontin (OPN), resistin, matrix metalloproteinase 7 (MMP7) and interferon ? (IFN?) as a four-marker panel while the addition of either CRP or adiponectin (ACRP-30) yielded comparable results in five protein signatures. Conclusions Proteins in the serum of CAD patients predominantly reflected (1) a positive acute phase, inflammatory response and (2) alterations in lipid metabolism, transport, peroxidation and accumulation. There were surprisingly few indicators of growth factor activation or extracellular matrix remodeling in the serum of CAD patients except for elevated OPN. These data suggest that many symptomatic patients without significant CAD could be identified by a targeted multiplex serum protein test without cardiac catheterization thereby eliminating exposure to ionizing radiation and decreasing the economic burden of angiographic testing for these patients.



Mathematical modeling as accounting: Predicting the fate of serum proteins and therapeutic monoclonal antibodies  

Microsoft Academic Search

This article reviews current efforts to mathematically model the half lives of serum proteins, especially antibodies. While it is recognized that the neonatal Fc receptor, FcRn, is necessary for longer serum persistence of certain proteins, particularly the high abundance IgGs and albumin, it is not clear that it is sufficient to completely determine the half lives of these proteins. More

Brian Gurbaxani



Observation of serum proteins in the edematous brain tissue by fluorescein-antibody technique  

Microsoft Academic Search

Histologic distribution of serum proteins within normal and edematous brain tissues was investigated with the fluorescein-antibody technique. Brain edema was produced in dogs with cold injury, as described byKlatzoet al. In normal brain large amounts of serum proteins were restricted to the vascular lumina and the presence of these proteins was sparse in nerve, glia and endothelial cells. On the

K. Someda; N. Kageyama



Extraction of C-reactive protein from serum on a microfluidic chip  

Microsoft Academic Search

The first extraction of a protein, C-reactive protein (CRP), from unadulterated serum on a microfluidic chip is demonstrated. Two stationary phases were evaluated for their ability to selectively extract this protein directly from serum without the need for additional cleanup steps. The first extraction media tested was an affinity matrix containing monoclonal antibodies to CRP, however, after immobilization this media

Michael G. Roper; Megan L. Frisk; Janeen P. Oberlander; Jerome P. Ferrance; Brian J. McGrory; James P. Landers



Proteome analysis of rat serum proteins adsorbed onto synthetic octacalcium phosphate crystals  

Microsoft Academic Search

The present study was designed to determine which proteins are selectively adsorbed onto two bone substitute materials, octacalcium phosphate (OCP) and hydroxyapatite (HA) crystals, from rat serum by proteome analysis. Ground crystals of synthetic OCP and commercially available sintered HA, with the same surface area, were incubated in rat serum proteins at 37°C for 24h. The proteins from the crystals

Hirofumi Kaneko; Junichi Kamiie; Hirotaka Kawakami; Takahisa Anada; Yoshitomo Honda; Naru Shiraishi; Shinji Kamakura; Tetsuya Terasaki; Hidetoshi Shimauchi; Osamu Suzuki



A Study of the Adhesive Glycoprotein-Inactivating Protein from Mammalian Blood Serum  

Microsoft Academic Search

A protein with a molecular weight of 70 kDa was isolated from bovine blood serum and purified to a homogenous state. This protein reversibly inhibited the adhesive serum glycoprotein with a molecular weight of 12 kDa, which displayed biological activity at ultralow doses. Amino acid analysis showed that the protein inactivator belongs to the group of prealbumins from vertebrate blood

V. P. Yamskova; E. Yu. Rybakova; A. A. Vinogradov; V. V. Vecherkin; I. A. Yamskov



Determination of Angiogenic Factors in Serum by Protein Array in Patients with Renal Cell Carcinoma  

Microsoft Academic Search

Using the protein array method we deter- mined the serum levels of a number of angiogenic factors. We identified serum levels of angiogenin, PDGF and MCP-1 (CCL2 chemokine) in serum of 32 patients with RCC, and 14 healthy volunteers by means of antibody array analysis. The patients were divided into three groups according to their disease stages (I+II, III, and



Prion protein detection in serum using micromechanical resonator arrays.  


Prion proteins that have transformed from their normal cellular counterparts (PrP(c)) into infectious form (PrP(res)) 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 desirable to develop non-invasive and ante mortem tests for the detection of prion proteins in bovine samples. Such ante mortem tests of all cows prior to slaughter will help to prevent the introduction of PrP(res) into the human food supply. Furthermore, detection of PrP(res) in donated blood will also help to prevent the transmission of CJD among humans through blood transfusion. In this study, we have continued development of a micromechanical resonator array that is capable of detecting PrP(c) in bovine blood serum. The sensitivity of the resonators for the detection of PrP(c) is further enhanced by the use of secondary mass labels. A pair of antibodies is used in a sandwich immunoassay format to immobilize PrP(c) on the surface of resonators and attach nanoparticles as secondary mass labels to PrP(c). Secondary mass labeling is optimized in terms of incubation time to maximize the frequency shifts that correspond to the presence of PrP(c) on the surface of resonators. Our results show that a minimum of 200 pg mL(-1) of PrP(c) in blood serum can be detected using micromechanical resonator arrays. PMID:19836525

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



Serum protein profiling by SELDI mass spectrometry: detection of multiple variants of serum amyloid alpha in renal cancer patients  

Microsoft Academic Search

The molecular analysis of serum is an important field for the definition of potential diagnostic markers or disease-related protein alterations. Novel proteomic technologies such as the mass spectrometric-based surface-enhanced laser desorption\\/ionization (SELDI) ProteinChip® technique facilitate a rapid and reproducible analysis of such protein mixtures and affords the researcher a new dimension in the search for biomarkers of disease. Here, we

Jonathan Tolson; Ralf Bogumil; Elke Brunst; Hermann Beck; Raimund Elsner; Andreas Humeny; Hartmut Kratzin; Martin Deeg; Markus Kuczyk; Gerhard A Mueller; Claudia A Mueller; Thomas Flad




PubMed Central

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

Jameson, Eloise; Roberts, Dorothy Brown



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.

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




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.

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



Prostate cancer serum biomarker discovery through proteomic analysis of alpha-2 macroglobulin protein complexes  

PubMed Central

Alpha-2 macroglobulin (A2M) functions as a universal protease inhibitor in serum and is capable of binding various cytokines and growth factors. In this study, we investigated if immunoaffinity enrichment and proteomic analysis of A2M protein complexes from human serum could improve detection of biologically relevant and novel candidate protein biomarkers in prostate cancer. Serum samples from six patients with androgen-independent, metastatic prostate cancer and six control patients without malignancy were analyzed by immunoaffinity enrichment of A2M protein complexes and MS identification of associated proteins. Known A2M substrates were reproducibly identified from patient serum in both cohorts, as well as proteins previously undetected in human serum. One example is heat shock protein 90 alpha (HSP90?), which was identified only in the serum of cancer patients in this study. Using an ELISA, the presence of HSP90? in human serum was validated on expanded test cohorts and found to exist in higher median serum concentrations in prostate cancer (n = 18) relative to control (n = 13) patients (median concentrations 50.7 versus 27.6 ng/mL, respectively, p = 0.001). Our results demonstrate the technical feasibility of this approach and support the analysis of A2M protein complexes for proteomic-based serum biomarker discovery.

Burgess, Earle F.; Ham, Amy-Joan L.; Tabb, David L.; Billheimer, Dean; Roth, Bruce J.; Chang, Sam S.; Cookson, Michael S.; Hinton, Timothy J.; Cheek, Kristin L.; Hill, Salisha; Pietenpol, Jennifer A.



Production of recombinant proteins in serum-free media  

Microsoft Academic Search

The advantages of serum-free culture for the manufacture of recombinant biopharmaceuticals from mammalian cells are reviewed. The process favoured is fed-batch serum-free cell culture. This process is applicable to the majority of cell lines, is practical on the large scale, gives the lowest manufacturing cost, and can b e carried out without the use of any serum.

D. Broad; R. Boraston; M. Rhodes



A Threshold for Central T Cell Tolerance to an Inducible Serum Protein1  

Microsoft Academic Search

We report an inducible system of self Ag expression that examines the relationship between serum protein levels and central T cell tolerance. This transgenic approach is based on tetracycline-regulated expression of a secreted form of hen egg lysozyme, tagged with a murine hemoglobin (Hb) epitope. In the absence of the tetracycline-regulated transactivator, serum levels of the chimeric protein are extremely

Dipica Haribhai; Deborah Engle; Michelle Meyer; David Donermeyer; J. Michael White; Calvin B. Williams


Serum bone Gla protein as a marker of bone turnover in acromegaly  

Microsoft Academic Search

Serum bone Gla protein, a sensitive and specific marker of bone turnover, was measured in 35 acromegalic patients (14 untreated, 8 clinically active, and 13 cured) and 21 controls. We also examined 10 acromegalic patients before and after transsphenoidal surgery. Untreated and clinically active acromegalic patients had significantly higher serum bone Gla protein concentrations than the control subjects. Other nonspecific

M. Marazuela; B. Astigarraga; M. J. Tabuenca; J. Estrada; F. Marín; T. Lucas



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

Microsoft Academic Search

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

A. R. Bradwell



Serum bone Gla protein as an indicator of parathyroidectomy in patients with secondary hyperparathyroidism  

Microsoft Academic Search

Bone Gla protein (BGP) is a vitamin K-dependent protein which is a marker of bone turnover. To determine whether serum BGP is a useful indicator for parathyroidectomy in patients with secondary hyperparathyroidism, we measured serum BGP levels. Thirty-seven patients with secondary hyperparathyroidism who were followed up for more than 1 year after parathyroidectomy were studied. All patients underwent total parathyroidectomy

Hiroshi Takami; Jun-ichi Shikata



Effect of serum proteins on osteoblast adhesion to surface-modified bioactive glass and hydroxyapatite.  


Previous studies indicate that modification of the surface of porous bioactive glass promotes osteoblast function. We hypothesize that bone formation on treated bioactive glass is due to the selective adsorption of serum attachment proteins. To test this hypothesis, we examined the profile of proteins adsorbed to treated bioactive glass and compared these proteins with those adsorbed to untreated bioactive glass and porous hydroxyapatite. Porous bioactive glass was treated with Tris-buffered electrolyte solution to generate a calcium phosphate-rich surface layer and then immersed in tissue-culture medium containing 10% serum. Proteins adsorbed to the ceramic surfaces were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot analysis. Porous hydroxyapatite bound a higher amount of total protein than did the other substrates. However, surface-modified porous bioactive glass adsorbed more fibronectin than did hydroxyapatite. The effect of serum-protein adsorption on osteoblast adhesion to bioactive glass and hydroxyapatite was also evaluated. Cell adhesion to porous bioactive glass that was surface-modified and serum-treated was significantly greater than to porous bioactive glass that was either surface-modified or serum-treated. Furthermore, cell adhesion to porous bioactive glass treated to form the dual layer of calcium phosphate and serum protein was significantly higher than adhesion to porous hydroxyapatite with adsorbed serum protein. Results of the study strongly suggest that adsorption of serum fibronectin to the surface of modified porous bioactive glass coated with calcium phosphate may be responsible for enhanced osteoblast adhesion. PMID:10376721

El-Ghannam, A; Ducheyne, P; Shapiro, I M



Serum alpha feto-protein screening in high risk pregnancies.  


We assayed alpha fetoprotein (AFP) in serum samples from 2,735 women during 14 to 20 weeks of gestation. Serum AFP levels were elevated in the presence of neural tube defect (NTD) and gut atresia in fetus, twin pregnancies, preterm delivery and neonatal complications. In two of the 23 cases of fetal NTD the diagnosis was suggested by AFP assays, with apparently normal ultrasound findings. Low maternal serum AFP levels were associated with chromosomal abnormalities and hydatidiform molar pregnancy. Calculation of risk for Down syndrome based on maternal serum AFP and maternal age helped to reduce the number of women requiring amniocentesis. Maternal serum AFP assay was helpful in the management of threatened abortion, suspected intrauterine death, and maternal toxoplasma infection. In seven cases where maternal serum AFP was high but ultrasound studies were normal, male babies were delivered. Thus maternal serum AFP assay proved useful in narrowing down the group of women requiring more detailed surveillance and diagnostic studies. PMID:10829851

Kaur, M; Verma, I C



Early In vitro Lipopolysaccharide-Induced Serum Protein Aggregation in Tolerant Serum.  

National Technical Information Service (NTIS)

In a rat LPS shock model, an intravenous injection of LPS (3 - 30 mg/kg body weight) corresponds to an estimated peak load of 0.13-1.3 mg LPS/ml serum. We tested the effects of such LPS concentrations in vitro. Rat serum was incubated with LPS (0.1 - 2.0 ...

M. A. Fletcher T. K. Morrison T. J. Williams



Design of Recombinant Antibody Microarrays for Serum Protein Profiling:  Targeting of Complement Proteins  

Microsoft Academic Search

Antibody-based microarrays is a novel technology with great promise for high-throughput proteomics. The process of designing high-performing arrays has, however, turned out to be challenging. Here, we have designed the next generation of a human recombinant scFv antibody microarray platform for protein expression profiling of nonfractionated biotinylated human plasma and serum proteomes. The setup, based on black polymer Maxisorb slides

Johan Ingvarsson; Anette Larsson; Anders G. Sjöholm; Lennart Truedsson; Bo Jansson; Carl A. K. Borrebaeck; Christer Wingren




PubMed Central

1. In view of the markedly short period of gestation in the dog and in view of the relatively large litters that are cast and nursed, this species when compared with the human being undergoes a much greater physiological strain during pregnancy and lactation. This is evidenced by marked decreases in the hematocrit values, in total cell volumes and in the serum protein concentrations, by an appreciable plasma hydration, and in some cases by significant reductions in the total circulating serum protein. 2. When pregnant dogs are fed a protein-free diet at a high level of caloric intake and are subjected to our standardized plasmapheresis technique, it is possible to deplete the animal of its reserve serum protein stores and reduce the serum protein concentration to the basal level (3.5 to 4.2 per cent) within the extremely short period of from 2 to 3 days. This indicates that the dog during pregnancy possesses a very limited amount of reserve serum protein. 3. Once the basal serum protein level is attained, the pregnant or lactating dog exhibits a marked impairment in its ability to regenerate serum protein. The synthesis of body proteins in the fetus during pregnancy and the milk proteins during lactation is considered to be actually an internal plasmapheresis, leading to a depletion of the serum protein by the preferential utilization of the materials from which this complex is made. These parasitic effects on the maternal organism are believed to be of primary importance, over and above any hydremia, in causing the lowered serum protein concentrations characteristic of pregnancy.

Melnick, Daniel; Cowgill, George R.



Evaluation of three high abundance protein depletion kits for umbilical cord serum proteomics  

PubMed Central

Background High abundance protein depletion is a major challenge in the study of serum/plasma proteomics. Prior to this study, most commercially available kits for depletion of highly abundant proteins had only been tested and evaluated in adult serum/plasma, while the depletion efficiency on umbilical cord serum/plasma had not been clarified. Structural differences between some adult and fetal proteins (such as albumin) make it likely that depletion approaches for adult and umbilical cord serum/plasma will be variable. Therefore, the primary purposes of the present study are to investigate the efficiencies of several commonly-used commercial kits during high abundance protein depletion from umbilical cord serum and to determine which kit yields the most effective and reproducible results for further proteomics research on umbilical cord serum. Results The immunoaffinity based kits (PROTIA-Sigma and 5185-Agilent) displayed higher depletion efficiency than the immobilized dye based kit (PROTBA-Sigma) in umbilical cord serum samples. Both the PROTIA-Sigma and 5185-Agilent kit maintained high depletion efficiency when used three consecutive times. Depletion by the PROTIA-Sigma Kit improved 2DE gel quality by reducing smeared bands produced by the presence of high abundance proteins and increasing the intensity of other protein spots. During image analysis using the identical detection parameters, 411 ± 18 spots were detected in crude serum gels, while 757 ± 43 spots were detected in depleted serum gels. Eight spots unique to depleted serum gels were identified by MALDI- TOF/TOF MS, seven of which were low abundance proteins. Conclusions The immunoaffinity based kits exceeded the immobilized dye based kit in high abundance protein depletion of umbilical cord serum samples and dramatically improved 2DE gel quality for detection of trace biomarkers.



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


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

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




PubMed Central

1. When the dog is subjected to quantitative plasmapheresis and fed appropriate "synthetic" artificial rations, it is possible to evaluate the ability of the organism to regenerate serum protein from both exogenous and endogenous sources. Approximately 44 per cent of the protein, casein, fed in excess of the minimal amount needed normally to meet the general nitrogen requirements, is utilized for the formation of new serum protein. Under our experimental conditions the dog can regenerate each week solely from endogenous sources approximately 0.6 gm. of this blood protein per kilo of optimal body weight. This is equivalent to about 21 per cent of the total amount of serum protein normally present in the plasma. 2. When the dog is fed an adequate protein diet and is subjected to a régime of prolonged intensive plasmapheresis (period of 16 consecutive weeks), no impairment in the ability of the organism to regenerate serum protein from either exogenous or endogenous sources occurs. Under our conditions of experimentation the dog appears to be able to form each week an amount of the blood protein approximately equal to that normally present in the plasma. Because of this remarkable ability of the normal organism to regenerate rapidly serum protein over a relatively long period of time, it seems that loss of protein alone in conditions of Bright's disease cannot be the etiologic agent responsible for the persistence of the hypoproteinemia. An additional factor, the "specific" ability of such individuals to regenerate serum protein, must be taken into consideration.

Melnick, Daniel; Cowgill, George R.



Clinical impact of serum proteins on drug delivery.  


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

Kratz, Felix; Elsadek, Bakheet



Simple colorimetry of glycated serum protein in a centrifugal analyzer.  


A recently described (Clin Chim Acta 127: 87-95, 1982) colorimetric assay for glycated proteins in serum exploits their ketoamine activity to reduce 3,3'-(3,3'-dimethoxy-4,4'-biphenylylene)bis [2-(p-nitrophenyl)-5-phenyl-2H-tetrazolium chloride] (nitroblue tetrazolium) in alkaline solution; the authors termed this the "fructosamine assay." The method is simple, requiring only the addition of one reagent and measurement of the absorbance change during 5 min; results are expressed relative to a synthetic standard. We have adapted this for use in a centrifugal analyzer and report its performance, both analytically and as an index of hyperglycemia. Precision is good (between-batch CV 2.1%), the reagent is stable and inexpensive, and the procedure is rapid (75 samples per hour). Albumin influences the measurement, but for concentrations greater than or equal to 35 g/L this was not a serious problem. Normal and diabetic populations can be clearly discriminated (p less than 0.001). The test detected 25 (84%) of the 30 untreated diabetics studied and gave four false positives (8%). The results correlate well with those for glucose in plasma of fasting subjects (r = 0.87) and for hemoglobin A1 (r = 0.80). PMID:6478601

Lloyd, D; Marples, J



Acute phase proteins and peritoneal dialysate albumin loss are the main determinants of serum albumin in peritoneal dialysis patients  

Microsoft Academic Search

Hypoalbuminemia predicts mortality in dialysis patients. It has been postulated that hypoalbuminemia in the dialysis population is a consequence of poor protein intake resulting from inadequate dialysis. To establish the cause of hypoalbuminemia in a group of 27 patients on peritoneal dialysis (PD), we determined the relationship between serum albumin concentration and a group of parameters including dialysis dose delivered

Jane Y. Yeun; George A. Kaysen



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


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

Wells, Beth; Innocent, Giles T; Eckersall, Peter D; McCulloch, Eilidh; Nisbet, Alasdair J; Burgess, Stewart T G



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

PubMed Central

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



A proteomic workflow for discovery of serum carrier protein-bound biomarker candidates of alcohol abuse using LC-MS/MS.  


The diagnosis and care of patients with alcohol abuse and dependence is hampered by a lack of sensitive and specific screening and monitoring tests. Proteomics is a good approach to search for biomarkers of alcohol abuse. Serum carrier protein-bound proteins have attracted significant interest because they remain a relatively un-mined region of the proteome. In the present study, a proteomic workflow including LC-MS/MS with enrichment of serum carrier protein-bound biomarkers technique was applied to profile the changes in quality and quantity of serum carrier protein-bound proteins for the discovery of novel biomarker candidates of alcohol abuse. In total, 311 proteins identified with high confidence were discovered to be bound to serum carrier proteins. Complement isoforms, Ig fragments, and apolipoprotein family proteins are the main serum carrier-bound proteins. Protein quantification analysis with and without concern as to gender revealed that gender is a critical consideration for biomarker development in alcohol abuse. Identified proteins not previously associated with alcohol abuse include gelsolin, selenoprotein P, serotransferrin, tetranectin, hemopexin, histidine-rich glycoprotein, plasma kallikrein, and vitronectin. Altered abundance of these proteins suggests that they may be potential novel biomarkers for alcohol abuse. PMID:19544491

Lai, Xianyin; Liangpunsakul, Suthat; Crabb, David W; Ringham, Heather N; Witzmann, Frank A



A Proteomic Workflow for Discovery of Serum Carrier Protein-Bound Biomarker Candidates of Alcohol Abuse Using Liquid Chromatography - Tandem Mass Spectrometry  

PubMed Central

The diagnosis and care of patients with alcohol abuse and dependence is hampered by a lack of sensitive and specific screening and monitoring tests. Proteomics is a good approach to search for biomarkers of alcohol abuse. Serum carrier protein-bound proteins have attracted significant interest because they remain a relatively un-mined region of the proteome. In the present study, a proteomic workflow including LC-MS/MS with enrichment of serum carrier protein-bound biomarkers technique was applied to profile the changes in quality and quantity of serum carrier protein-bound proteins for the discovery of novel biomarker candidates of alcohol abuse. In total, 311 proteins identified with high confidence were discovered to be bound to serum carrier proteins. Complement isoforms, Ig fragments, and apolipoprotein family proteins are the main serum carrier-bound proteins. Protein quantification analysis with and without concern as to gender revealed that gender is a critical consideration for biomarker development in alcohol abuse. Identified proteins not previously associated with alcohol abuse include gelsolin, selenoprotein P, serotransferrin, tetranectin, hemopexin, histidine-rich glycoprotein, plasma kallikrein, and vitronectin. Altered abundance of these proteins suggests that they may be potential novel biomarkers for alcohol abuse.

Lai, Xianyin; Liangpunsakul, Suthat; Crabb, David W.; Ringham, Heather N.; Witzmann, Frank A.



Serum heat shock protein 70 levels, oxidant status, and mortality in sepsis.  


Animal studies as well as prospective randomized clinical trials associated sepsis with redox imbalance and oxidative stress, but other studies failed to establish a correlation between antioxidant-based therapies and improvement of sepsis condition. This is also true for studies on the role of the chaperone heat shock protein 70 (HSP70), which is increased in serum during sepsis. Heat shock protein 70 is affected at several levels by oxidative stress, but this relationship has never been studied in sepsis. Here, we evaluated the relationship between serum HSP70 immunocontent and oxidant status in sepsis. Patients with severe sepsis were followed up for 28 days after diagnosis, or until death. Up to a maximum of 12 h after sepsis diagnosis, serum was collected for determination of HSP70 immunocontent by Western blot and evaluation of oxidative parameters (TRAP [total radical-trapping antioxidant parameter], TBARSs [thiobarbituric acid-reactive substances], and carbonyl levels). Serum of sepsis patients presented enhanced HSP70 levels. Analysis of oxidative parameters revealed that septic patients with pronounced oxidative damage in serum had also increased HSP70 serum levels. Sepsis patients in whom serum oxidative stress markers were not different from control presented normal serum HSP70. Analysis of septic patients according to survival outcome also indicated that patients with increased HSP70 serum levels presented increased mortality. We concluded that serum HSP70 levels are modulated according to the patient oxidant status, and increased serum HSP70 is associated to mortality in sepsis. PMID:21330950

Gelain, Daniel Pens; de Bittencourt Pasquali, Matheus Augusto; M Comim, Clarissa; Grunwald, Marcelo Sartori; Ritter, Cristiane; Tomasi, Cristiane Damiani; Alves, Sarah Cascaes; Quevedo, Joao; Dal-Pizzol, Felipe; Moreira, José Cláudio Fonseca



Feasibility study of gel filtration as a separation method for blood serum proteins in combination with PIXE analysis  

Microsoft Academic Search

The combination between a protein separation technique and the PIXE method has a great potential for large surveys, including\\u000a thousands of samples, in which multielemental analysis is required. Gel filtration with a Sephadex G-200 gel and a TRIS-acetate\\u000a buffer was used for separating proteins in human serum. The fractions were doped with an yttrium\\/vanadium standard and then\\u000a concentrated and pipeted

Jan Pallon; Pirjo Pakarinen; Klas Malmqvist; K. Roland Akselsson



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

PubMed Central

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

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



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.

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.



Heat Shock Protein 70 Serum Levels Differ Significantly in Patients with Chronic Hepatitis, Liver Cirrhosis, and Hepatocellular Carcinoma  

PubMed Central

Members of the heat shock protein 70 (HSP70) family play an important role in assisting protein folding, preventing protein aggregation and transport of proteins across membranes under physiological conditions. Following environmental (i.e., irradiation, chemotherapy), physiological (i.e., cell growth, differentiation), and pathophysiological (i.e., inflammation, tumorigenesis) stress, the synthesis of heat shock proteins (HSPs) is highly up-regulated, whereas protein synthesis in general is reduced. In contrast to normal cells, many tumor entities including hepatocellular carcinoma (HCC) overexpress HSP70, the major-stress-inducible member of the HSP70 family, present it on their cell surface and secrete it into the extracellular milieu. Herein, the prognostic relevance of serum HSP70 levels in patients with chronic hepatitis (CH; n?=?50), liver cirrhosis (LC; n?=?46), and HCC (n?=?47) was analyzed. Similar to other tumor entities, HSP70 is also present on the surface of primary HCC cells. The staining intensity of intracellular HSP70 in HCC tissue is stronger compared to control and cirrhotic liver sections. HSP70 serum levels in all HCC patients were significantly higher compared to a control group without liver disease (n?=?40). No significant age- and gender-related differences in HSP70 serum levels were observed in male and female healthy human volunteers (n?=?86). Patients with CH (n?=?50) revealed significantly higher HSP70 serum levels compared to the control group, however, these values were significantly lower than those of HCC patients (n?=?47). Furthermore, a subgroup of patients with LC who subsequently developed HCC (LC-HCC, n?=?13) revealed higher HSP70 serum levels than patients with LC (n?=?46, p?=?0.05). These data indicate that serum HSP70 levels are consecutively increased in patients with CH, LC and liver carcinomas and thus might have a prognostic value.

Gehrmann, Mathias; Cervello, Melchiorre; Montalto, Giuseppe; Cappello, Francesco; Gulino, Alessandro; Knape, Clemens; Specht, Hanno M.; Multhoff, Gabriele



Detection of serum proteins by native polyacrylamide gel electrophoresis using Blue Sepharose CL6B-containing stacking gels  

Microsoft Academic Search

Analysis of serum proteins by native polyacrylamide gel electrophoresis is difficult because albumin is abundant in serum and interferes with the resolution of other proteins, especially ?-antitrypsin which has mobility that is very similar to that of albumin. We present here a method in which serum proteins are separated by polyacrylamide gel electrophoresis using stacking gels containing Blue Sepharose CL-6B,

Haruhiro Muratsubaki; Kaoru Satake; Yasuhisa Yamamoto; Keiichiro Enomoto



Relation of Bone Marrow Findings to Serum Protein Changes in Lymphosarcoma, Chronic Lyinphocytic Leukemia and Hodgkin's Disease  

Microsoft Academic Search

LTERATIONS in serum protein demonstrable by electrophoretic analy- sis have been noted fairly frequently in cases of lymphosarcoma, lym- phocytic leukemia and Hodgkin's disease. Abnormal proteins such as cryo- globulins and myeloma-type proteins also have been observed in the serum of an occasional patient with these conditions. in this study we attempted to determine whether disturbances in serum globulins are




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

Microsoft Academic Search

Fatty acid binding protein (FABP) 4 chaperones free fatty acids (FFAs) in the adipocytes during lipolysis. Serum FFA relates to metabolic syndrome, and serum FABP4 is emerging as a novel risk marker. In 36 overweight\\/obese women, serum FABP4 and FFA were measured hourly during 5-hour oral glucose tolerance test. Insulin resistance was determined using frequently sampled intravenous glucose tolerance test.

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



Ovine lentivirus antibody detection in serum, colostrum and milk using a recombinant transmembrane protein ELISA  

Microsoft Academic Search

An enzyme-linked immunosorbent assay (ELISA) was used to detect antibodies against ovine lentivirus (OLV) in serum, colostrum, and milk from naturally infected sheep. The assay used OLV recombinant transmembrane envelope protein (rTM) as a test antigen. Matched serum\\/colostrum and serum\\/milk samples were collected at 24h, 4weeks (mid-lactation), and 8 weeks (weaning) post-lambing. Among 129 paired samples collected at 24 h

Jim Keen; Jimmy Kwang; E. Travis Littledike; Laura L. Hungerford



Induction of Hepatic Synthesis of Serum Amyloid A Protein and Actin  

Microsoft Academic Search

Major changes in the mRNA population of murine liver occur after administration of bacterial lipopolysaccharide, an agent that causes increases in the concentrations of acutephase serum proteins. The mRNA for one of these, serum amyloid A, is increased at least 500-fold compared to the normal level. It becomes one of the most abundant hepatic mRNAs, and serum amyloid A synthesis

John F. Morrow; Robert S. Stearman; Cynthia G. Peltzman; David A. Potter



Chickpea protein hydrolysate as a substitute for serum in cell culture  

Microsoft Academic Search

The growth of mammalian cells in vitro requires the use of rich culture media that are prepared by combining serum with specific\\u000a nutrient formulations. Serum, the most expensive component of culture media, provides a complex mixture of growth factors\\u000a and nutrients. Protein hydrolysates that can support in vitro cell growth and eliminate or reduce the need to use serum have\\u000a been obtained

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



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.



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.




PubMed Central

The detection of recombinant human growth hormone (rhGH) is difficult due to its short half-life; therefore, novel and robust biomarkers of rhGH abuse are needed. In this study, serum samples derived from subjects treated with rhGH in a randomized, double blind, placebo-controlled crossover study were analyzed by 2-DE coupled with MS. Eight healthy male subjects aged 23.2 ± 0.6 yr were injected with rhGH (2 mg/day) or saline for 7 days with serum samples drawn at days 0, 3, and 8. Protein intensities were quantified and analyzed for differences between rhGH versus placebo treatments. Protein that showed significant changes were identified and confirmed by Western blotting. These included specific isoforms of alpha-1 antitrypsin and transthyretin that increased; and inter-alpha-trypsin inhibitor heavy chain H4, apolipoprotein A-1 and hemoglobin beta chain that decreased. These proteins represent novel biomarkers of short-term rhGH exposure and may lead to a new method for detecting rhGH doping.

Ding, Juan; Okada, Shigeru; J?rgensen, Jens Otto Lunde; Kopchick, John J.



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  

Microsoft Academic Search

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 (D0)

Wei-Li Liao; Illarion V. Turko



Determination of Transaminase Activity of Serum Protein Fractions separated by Paper Electrophoresis  

Microsoft Academic Search

THE electrophoretic mobility of glutamic-oxalacetic acid transaminase in blood serum has been found to be between those for alpha2- and beta-globulins1. In our experiments2 the glutamic-oxalacetic acid transaminase was associated mainly with alpha2-globulins. This communication deals with this transaminase and glutamic-pyruvic acid transaminase and their relations to serum protein fractions.

Miroslav Sevela



Relationship between fructosamine with serum protein, albumin and glucose concentrations in dairy ewes  

Microsoft Academic Search

The aim of the study was to investigate the changes in blood fructosamine concentrations of ewes during late pregnancy and lactation. The relationship of serum fructosamine to changes in the serum glucose and blood protein and albumin concentrations were measured in 10 crossbred dairy ewes. Blood was sampled 10 days prior to lambing (D?10), on day 10 (D+10), day 20

N. Filipovi?; Z. Stojevi?; T. Mašek; Ž. Mikulec; N. Prvanovi?



Serum alpha feto-protein screening in high risk pregnancies  

Microsoft Academic Search

We assayed alpha fetoprotein (AFP) in serum samples from 2,735 women during 14 to 20 weeks of gestation. Serum AFP levels\\u000a were elevated in the presence of neural tube defect (NTD) and gut atresia in fetus, twin pregnancies, preterm delivery and\\u000a neonatal complications. In two of the 23 cases of fetal NTD the diagnosis was suggested by AFP assays, with

Manjeet Kaur; Ishwar C. Verma



Peptides capable of binding to serum proteins and compounds, constructs and polypeptides comprising the same  

US Patent & Trademark Office Database

The present invention relates to amino acid sequences that are capable of binding to serum proteins; to compounds, proteins, polypeptides, fusion proteins or constructs comprising or essentially consisting of such amino acid sequences; to nucleic acids that encode such amino acid sequences, compounds, proteins, polypeptides, fusion proteins or constructs; to compositions, and in particular pharmaceutical compositions, that comprise such amino acid sequences, compounds, proteins, polypeptides, fusion proteins or constructs; and to uses of such amino acid sequences, compounds, proteins, polypeptides, fusion proteins or constructs.



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.

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



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)



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


When nanoparticles (NP) enter the body they come into contact with body fluids containing proteins which can adsorb to their surface. These proteins may influence the NP interactions with the biological vicinity, eventually determining their biological fate inside the body. Adsorption of the most abundantly binding proteins was studied after an in vitro 24 hr incubation of monodisperse, negatively charged 5, 15 and 80 nm gold spheres (AuNP) in mouse serum by a two-step analysis: proteomic protein identification and quantitative protein biochemistry. The adsorbed proteins were separated from non-adsorbed proteins by centrifugation and gel electrophoresis and identified using a MALDI-TOF-MS-Proteomics-Analyzer. Quantitative analysis of proteins in gel bands by protein densitometry, required the focus on predominantly binding serum proteins. Numerous proteins adsorbed to the AuNP depending on their size, e.g., apolipoproteins or complement C3. The qualitative and quantitative amount of adsorbed proteins differed between 5, 15 and 80 nm AuNP. Band intensities of adsorbed proteins decreased with increasing AuNP sizes based not only on their mass but also on their surface area. Summarizing, the AuNP surface is covered with serum proteins containing transport and immune related proteins among others. Hence, protein binding depends on the size, surface area and curvature of the AuNP. PMID:23735821

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



A Comparative Study of Cross Reactions of Serum Protein Antigens of Seven Mammals.  

National Technical Information Service (NTIS)

The study on the antigenic cross reactions among the serum proteins of mammals is not only definitely important in bio-taxonomy, embryology, and evolutional theory, but also important for immunobiological and immunochemical researches. If a cross reaction...

Y. Hsieh L. Shih S. Ch'en H. Jao C. Ch'ien



Serum Protein-Bound Carbohydrates and Other Glycoprotein Assays as Indicators of Tumor Burden.  

National Technical Information Service (NTIS)

Elevations in serum glycoproteins and protein-bound carbohydrates (PBC) are significant consequences of radiation damage, trauma, and certain disease states such as cancer. Thus, it is important to determine the diagnostic, prognostic, and functional sign...

J. F. Weiss W. P. Bradley A. P. Blasco J. C. Alexander N. A. Silverman



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


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

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



Deglycosylation of Serum Vitamin D3-binding Protein Leads to Immunosuppression in Cancer Patients1  

Microsoft Academic Search

Serum vitamin D,-binding protein (Gc protein) can be converted by Ăź-galactosidaseof B cells and sialidase of T cells to a potent macrophage activating factor, a protein with jV-acetylgalactosamine as the remaining sugar moiety. Thus, Gc protein is the precursor of the macrophage activating factor iMAr'i. Treatment of Gc protein with immobilized Ăź-ga- lactosidase and sialidase generates an extremely high titered

Nobuto Yanulinolo; Venkateswara R. Naraparaju; Sucha O. Asbell



Increased serum heat-shock protein 70 levels reflect systemic inflammation, oxidative stress and hepatocellular injury in preeclampsia.  


It has been previously reported that serum levels of 70-kDa heat-shock protein (Hsp70) are elevated in preeclampsia. The aim of the present study was to examine whether increased serum Hsp70 levels are related to clinical characteristics and standard laboratory parameters of preeclamptic patients, as well as to markers of inflammation (C-reactive protein), endothelial activation (von Willebrand factor antigen) or endothelial injury (fibronectin), trophoblast debris (cell-free fetal DNA) and oxidative stress (malondialdehyde). Sixty-seven preeclamptic patients and 70 normotensive, healthy pregnant women were involved in this case-control study. Serum Hsp70 levels were measured with enzyme-linked immunosorbent assay (ELISA). Standard laboratory parameters (clinical chemistry) and C-reactive protein (CRP) levels were determined by an autoanalyzer using the manufacturer's kits. Plasma von Willebrand factor antigen (VWF:Ag) levels were quantified by ELISA, and plasma fibronectin concentration by nephelometry. The amount of cell-free fetal DNA in maternal plasma was determined by quantitative real-time polymerase chain reaction analysis of the sex-determining region Y gene. Plasma malondialdehyde levels were measured by the thiobarbituric acid-based colorimetric assay. Serum Hsp70 levels were increased in preeclampsia. Furthermore, serum levels of blood urea nitrogen, creatinine, bilirubin and CRP, serum alanine aminotransferase and lactate dehydrogenase (LDH) activities, as well as plasma levels of VWF:Ag, fibronectin, cell-free fetal DNA and malondialdehyde were also significantly higher in preeclamptic patients than in normotensive, healthy pregnant women. In preeclamptic patients, serum Hsp70 levels showed significant correlations with serum CRP levels (Spearman R = 0.32, p = 0.010), serum aspartate aminotransferase (R = 0.32, p = 0.008) and LDH activities (R = 0.50, p < 0.001), as well as with plasma malondialdehyde levels (R = 0.25, p = 0.043). However, there was no other relationship between serum Hsp70 levels and clinical characteristics (age, parity, body mass index, blood pressure, gestational age, fetal birth weight) and laboratory parameters of preeclamptic patients, including markers of endothelial activation or injury and trophoblast debris. In conclusion, increased serum Hsp70 levels seem to reflect systemic inflammation, oxidative stress and hepatocellular injury in preeclampsia. Nevertheless, further studies are required to determine whether circulating Hsp70 plays a causative role in the pathogenesis of the disease. PMID:18686014

Molvarec, Attila; Rigó, János; Lázár, Levente; Balogh, Krisztián; Makó, Veronika; Cervenak, László; Mézes, Miklós; Prohászka, Zoltán



Increased serum heat-shock protein 70 levels reflect systemic inflammation, oxidative stress and hepatocellular injury in preeclampsia  

PubMed Central

It has been previously reported that serum levels of 70-kDa heat-shock protein (Hsp70) are elevated in preeclampsia. The aim of the present study was to examine whether increased serum Hsp70 levels are related to clinical characteristics and standard laboratory parameters of preeclamptic patients, as well as to markers of inflammation (C-reactive protein), endothelial activation (von Willebrand factor antigen) or endothelial injury (fibronectin), trophoblast debris (cell-free fetal DNA) and oxidative stress (malondialdehyde). Sixty-seven preeclamptic patients and 70 normotensive, healthy pregnant women were involved in this case-control study. Serum Hsp70 levels were measured with enzyme-linked immunosorbent assay (ELISA). Standard laboratory parameters (clinical chemistry) and C-reactive protein (CRP) levels were determined by an autoanalyzer using the manufacturer’s kits. Plasma von Willebrand factor antigen (VWF:Ag) levels were quantified by ELISA, and plasma fibronectin concentration by nephelometry. The amount of cell-free fetal DNA in maternal plasma was determined by quantitative real-time polymerase chain reaction analysis of the sex-determining region Y gene. Plasma malondialdehyde levels were measured by the thiobarbituric acid-based colorimetric assay. Serum Hsp70 levels were increased in preeclampsia. Furthermore, serum levels of blood urea nitrogen, creatinine, bilirubin and CRP, serum alanine aminotransferase and lactate dehydrogenase (LDH) activities, as well as plasma levels of VWF:Ag, fibronectin, cell-free fetal DNA and malondialdehyde were also significantly higher in preeclamptic patients than in normotensive, healthy pregnant women. In preeclamptic patients, serum Hsp70 levels showed significant correlations with serum CRP levels (Spearman R?=?0.32, p?=?0.010), serum aspartate aminotransferase (R?=?0.32, p?=?0.008) and LDH activities (R?=?0.50, p?serum Hsp70 levels and clinical characteristics (age, parity, body mass index, blood pressure, gestational age, fetal birth weight) and laboratory parameters of preeclamptic patients, including markers of endothelial activation or injury and trophoblast debris. In conclusion, increased serum Hsp70 levels seem to reflect systemic inflammation, oxidative stress and hepatocellular injury in preeclampsia. Nevertheless, further studies are required to determine whether circulating Hsp70 plays a causative role in the pathogenesis of the disease.

Rigo, Janos; Lazar, Levente; Balogh, Krisztian; Mako, Veronika; Cervenak, Laszlo; Mezes, Miklos; Prohaszka, Zoltan



Maternal Serum Placental Protein 13 at Eleven to Thirteen Weeks in Chromosomally Abnormal Pregnancies  

Microsoft Academic Search

Objective: To investigate whether the maternal serum concentration of placental protein 13 (PP13) is altered in chromosomally abnormal pregnancies and to examine the potential value of this placental protein in screening for aneuploidies at 11–13 weeks. Methods: The maternal serum concentration of PP13 at 11–13 weeks was compared in 536 euploid and 134 aneuploid pregnancies (trisomy 21: n = 49;

Ranjit Akolekar; José María Pérez Penco; Evdoxia Skyfta; Jesús Rodríguez Calvo; Kypros H. Nicolaides



Low protein serum-free medium for antibody-production in stirred bioreactors  

Microsoft Academic Search

Summary A chemically defined serum-free medium was modified with respect to protein content by substituting PEG (polyethylen glycol) for BSA-FA (bovine serum albumin fatty acid complex). The outstanding advantage of this low-protein medium is the excellent support of hybridoma-growth and MAb (monoclonal antibody)-production in stirred bioreactors even when inoculated at low cell concentrations. Using this medium, the secreted MAbs were

Horst D. Blasey; Ursula Winzer



Increased serum S100B protein in schizophrenia: a study in medication-free patients  

Microsoft Academic Search

S100B protein, a calcium binding protein produced and released by glial cells, has been used as a sensitive marker of brain damage. Previous studies have found alterations in peripheral S100B levels in schizophrenic patients on medication. We compared serum S100B levels of 20 medication-free DSM-IV schizophrenic patients and 20 age-gender matched healthy controls. Schizophrenic patients presented higher serum S100B levels

D. R Lara; C. S. Gama; P. Belmonte-de-Abreu; L. V. C. Portela; C. A. Gonçalves; M. Fonseca; S. Hauck; D. O. Souza



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.



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


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



[Serum eosinophil cationic protein in children with allergic and nonallergic inflammation].  


Serum eosinophil cationic protein (ECP) levels increase in inflammation processes with activation of eosinophils. We studied serum ECP in (I) 32 pollinic children without symptoms, in June and October and (II) 10 children with acute asthma crisis. As control groups we included (III) 25 children sent to the hospital with suspected allergic diseases in which an IgE mediated process could be ruled out; (IV) 34 coeliac patients; (V) 15 children with cystic fibrosis and (VI) 48 normal children. The pollinic children had increased figures of ECP in June (21.2 +/- 9.2 micrograms/L) compared to normal controls (p < 0.001) and they continued to have high levels in October (13.5 +/- 9.2 micrograms/L, p < 0.05). The patients with very high ECP (> 20 micrograms/K), in spite of being asymptomatic, showed a negative correlation between ECP/peak-flow (p: 0.038). In addition, in these patients the ECP also had a negative correlation with the recovery of bronchospasm from June to October (p: 0.024). Some asthmatic children also had high ECP, but the results were too heterogeneous to draw any conclusions, possibly due to the drugs received. The ECP was independent of age and sex. It not correlated with serum IgE, nevertheless, in non-atopic patients it did correlate with blood eosinophilia (p < 0.005). In coeliac and cystic fibrosis patients, we did not find ECP to be increased. In conclusion, serum ECP increases in some allergic patients and suspected allergy, but not in all cases. It does not increase in other chronic mucosal inflammations, such as coeliac or cystic fibrosis. It correlates with bronchospasms and would have some value in predicting short-term evolution. PMID:8830600

Gómez Carrasco, J A; Blanco Quirós, A; Arranz Sanz, E; Tellería Orriols, J J; Lapeńa López de Armentia, S; Alvarez Mon, M



Water-in-carbon dioxide microemulsions: An environment for hydrophiles including proteins  

SciTech Connect

Carbon dioxide in the liquid and supercritical fluid states is useful as a replacement for toxic organic solvents. However, nonvolatile hydrophilic substances such as proteins, ions, and most catalysts are insoluble. This limitation was overcome by the formation of aqueous microemulsion droplets in a carbon dioxide-continuous phase with a nontoxic ammonium carboxylate perfluoropolyether surfactant. Several spectroscopic techniques consistently indicated that the properties of the droplets approach those of bulk water. The protein bovine serum albumin (BSA) with a molecular weight of 67,000 is soluble in this microemulsion and experiences an environment similar to that of native BSA in buffer. 23 refs., 4 figs.

Johnston, K.P.; Harrison, K.L. [Univ. of Texas, Austin, TX (United States); Clarke, M.J. [Univ. of Nottingham (United Kingdom)] [and others



Clinical efficacy of serum human epididymis protein 4 as a diagnostic biomarker of ovarian cancer: A pilot study  

PubMed Central

Objective To compare accuracy of serum human epididymis protein 4 (HE4) levels with cancer antigen 125 (CA-125) levels as biomarkers for ovarian cancer. Methods The study population included 94 Korean women, including 32 patients with a diagnosis of ovarian cancer and 62 patients with a diagnosis of benign ovarian tumor. All diagnoses were confirmed by histopathological analysis. Serum HE4 levels were assessed using an HE4 enzyme immunoassay, which were performed according to the manufacturer's instructions. Serum CA-125 levels were determined using a Modular analytics E170 module. Results The median serum CA-125 and HE4 levels were significantly higher in patients with ovarian cancer than those with other benign tumors (CA-125, 394.1 U/mL vs. 22.7 U/mL; HE4, 56.7 pM vs. 18.5 pM; P < 0.05 in both). An additional comparison revealed that the patients with endometriosis had greater median serum CA-125 levels than those with other benign ovarian tumors (32.0 U/mL vs. 17.9 U/mL, P = 0.03). Conversely, the median serum HE4 levels were similar among the two benign ovarian tumor groups, with no statistically significant difference observed (19.0 pM vs. 18.2 pM, P = 0.49). The receiver operating characteristics curve analysis for the benign ovarian tumor and ovarian cancer patients showed that HE4 showed a greater area under the curve with borderline significance when compared with CA-125 in both groups (0.93 vs. 0.85). Conclusion Serum HE4 levels may not only allow for the detection of ovarian cancer, but also allow for better differentiation of cases of ovarian cancer versus other benign ovarian tumors compared with serum CA-125.

Chung, Shin Hye; Lee, Soo Yoon; Ju, Woong



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


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



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


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



Effect of protein binding in serum on therapeutic efficacy of cephem antibiotics.  

PubMed Central

The effect of protein binding in serum of eight cephem antibiotics (ceftazidime, ceftizoxime, cefotiam, cefmetazole, cefpiramide, cefazolin, cefuzonam, ceftriaxone) on their therapeutic efficacies was examined in mice with experimentally induced intraperitoneal infections or pneumonia. The relationship among therapeutic activity, in vitro antibacterial activity, total or free (unbound) levels in serum, and homogenized whole lung levels was investigated. In the intraperitoneal infection caused by Staphylococcus aureus or Klebsiella pneumoniae, the 50% effective doses (ED50s) of the cephem antibiotics correlated with the area under the concentration-time curve (AUC) values of free levels in serum and the MICs but not with those of total levels in serum. A linear relationship was seen between 1/ED50 values and AUC of free levels in serum/MIC values. On the other hand, in mice with pneumonia caused by K. pneumoniae, the number of bacteria in the lung closely correlated with the AUC of the antibiotic concentration in lung tissue. There was a direct correlation between the levels in lung tissue and total levels in serum but not free levels in serum. The cephem antibiotics tested in this study were bound only slightly to homogenates of mouse lung. These results indicate that the effect of protein binding in serum on therapeutic efficacy against intraperitoneal infection differs from that against pulmonary infection. Images

Tawara, S; Matsumoto, S; Kamimura, T; Goto, S



Xenoestrogenicity in in vitro assays is not caused by displacement of endogenous estradiol from serum proteins.  


The possibility that compounds tested for estrogenicity can compete for binding places on serum proteins and cause an increase of available and very potent endogenous estrogens is of great interest for both the in vitro assay results and the prediction of risk for humans. In in vitro assays, small amounts of estradiol remaining after the charcoal stripping of serum applied in the culture medium could be displaced by the tested compounds, leading to an estrogenic response that might be falsely attributed to the test compound. We have studied the stripping efficiency of charcoal and measured whether reported xenoestrogens can displace estradiol from serum in an in vitro assay using negligible depletion-solid phase microextraction (nd-SPME). Possible competition was also studied with a mathematical exposure model, from which the predictions were compared to the measurements. We found that the common charcoal stripping procedure removed 99% of initially present estradiol. Additionally, our results with charcoal adsorption indicate that charcoal is not useful for serum protein binding assays, as it adsorbs more than the free fraction of ligand. Although the competition model predicted a displacement of estradiol from the serum proteins at the higher applied doses of xenoestrogen, the measurements showed no displacement. Therefore, we conclude that estrogenic responses in the in vitro assay applied here are not caused by displacement of remaining estradiol in the stripped serum. The possibility remains, however, that our displacement hypothesis does apply for estrogen sulfates, as these are present in much higher concentrations than estradiol in stripped serum. PMID:15282407

Heringa, Minne B; van der Burg, Bart; van Eijkeren, Jan C H; Hermens, Joop L M



Tetranectin-like protein in vertebrate serum: a comparative immunochemical analysis.  


The glycoprotein tetranectin (TN) found in human serum is a 90-kDa homotrimeric C-type lectin binding Ca2+, heparin and plasminogen kringle 4. TN is suggested as being implicated in tissue remodelling. The antigenic reactivity of putative TN was examined in serum from 14 different animal species using three sandwich enzyme immunoassays for human TN. Crab-eating macaque serum showed the strongest reaction, followed by horse and cat. Serum from cow, goat, pig, mouse and chicken reacted weakly, while dog, trout, and the amphibian and the reptile species did not react. The TN-like protein from macaque, horse and cat serum bound heparin and showed the same dependence on Ca2+ for interaction with the monoclonal antibodies as human TN. Gel filtration of sera from the three animal species showed that the TN-like protein eluted as single peaks with a M(r) of 70-90 kDa. Western blotting of horse and cat TN-like protein electrophoresed under reducing conditions showed that the antibodies against human TN reacted with a single band with an approximate M(r) of 30 kDa, indicating that the TN-like protein is also a homotrimer. Horse and cat TN-like protein interacted with human kringle 4-sepharose. Most likely, the reacting protein represents crab-eating macaque, horse and cat homologues of human TN. PMID:11290444

Thougaard, A V; Jaliashvili, I; Christiansen, M



Comparative protein profiling of serum and plasma using an antibody suspension bead array approach.  


In the pursuit towards a systematic analysis of human diseases, array-based approaches within antibody proteomics offer high-throughput strategies to discover protein biomarkers in serum and plasma. To investigate the influence of sample preparation on such discovery attempts, we report on a systematic effort to compare serum and plasma protein profiles determined with an antibody suspension bead array. The intensity levels were used to define protein profiles and no significant differences between serum and plasma were observed for 79% of the 174 antibodies (targeting 156 proteins). By excluding 36 antibodies giving rise to differential intensity levels, cluster analysis revealed donor-specific rather than preparation-dependent grouping. With a cohort from a clinically relevant medical condition, the metabolic syndrome, the influence of the sample type on a multiplexed biomarker discovery approach was further investigated. Independent comparisons of protein profiles in serum and plasma revealed an antibody targeting ADAMTSL-4, a protein that would qualify to be studied further in association with the condition. In general, the preparation type had an impact on the results of the applied antibody suspension bead array, and while the technical variability was equal, plasma offered a greater biological variability and allowed to give rise to more discoveries than serum. PMID:19953555

Schwenk, Jochen M; Igel, Ulrika; Kato, Bernet S; Nicholson, George; Karpe, Fredrik; Uhlén, Mathias; Nilsson, Peter



Systematic Discovery of Ectopic Pregnancy Serum Biomarkers Using 3-D Protein Profiling Coupled with Label-free Quantitation  

PubMed Central

Ectopic pregnancy (EP) and normal intrauterine pregnancy (IUP) serum proteomes were quantitatively compared to systematically identify candidate biomarkers. A 3-D biomarker discovery strategy consisting of abundant protein immunodepletion, SDS gels, LC-MS/MS, and label-free quantitation of MS signal intensities identified 70 candidate biomarkers with differences between groups greater than 2.5-fold. Further statistical analyses of peptide quantities were used to select the most promising 12 biomarkers for further study, which included known EP biomarkers, novel EP biomarkers (ADAM12 and ISM2), and five specific isoforms of the pregnancy specific beta-1-glycoprotein family. Technical replicates showed good reproducibility and protein intensities from the label-free discovery analysis compared favorably with reported abundance levels of several known reference serum proteins over at least three orders of magnitude. Similarly, relative abundances of candidate biomarkers from the label-free discovery analysis were consistent with relative abundances from pilot validation assays performed for five of the 12 most promising biomarkers using label-free multiple reaction monitoring of both the patient serum pools used for discovery and the individual samples that constituted these pools. These results demonstrate robust, reproducible, in-depth 3-D serum proteome discovery, and subsequent pilot-scale validation studies can be achieved readily using label-free quantitation strategies.

Beer, Lynn A.; Tang, Hsin-Yao; Sriswasdi, Sira; Barnhart, Kurt T.; Speicher, David W.



The Members of the Plakin Family of Proteins Recognized by Paraneoplastic Pemphigus Antibodies Include Periplakin  

Microsoft Academic Search

Sera of patients with paraneoplastic pemphigus (PNP) characteristically immunoprecipitate five proteins, observations confirmed with the sera examined in this study. The proteins characterized thus far as autoantigens in PNP all belong to the plakin family of proteins and include desmoplakin, the 230 kDa bullous pemphigoid antigen, and envoplakin. The pattern of bands precipitated from metabolically labeled human keratinocyte extracts by

My G. Mahoney; Sirpa Aho; Jouni Uitto; John R. Stanley



Abnormal protein patterns in blood serum and cerebrospinal fluid detected by capillary electrophoresis  

Microsoft Academic Search

Capillary zone electrophoresis and high-resolution agarose gel electrophoresis were compared to detect protein components in serum and cerebrospinal fluid of patients. Both electrophoretic methods proved to be useful for detection of protein abnormalities (e.g., mono- and oligoclonal bands) in biological fluids, but capillary electrophoresis offered several important advantages, such as sample application without preliminary concentration, lack of staining procedures, and

Mariella Ivanova; Elena Tzvetanova; Veska Jetcheva; Ferenc Kilár



Reproducibility of SELDI-TOF protein patterns in serum: comparing datasets from different experiments  

Microsoft Academic Search

Motivation: There has been much interest in using patterns derived from surface-enhanced laser desorption and ioniza- tion (SELDI) protein mass spectra from serum to differentiate samples from patients both with and without disease. Such patterns have been used without identification of the underly- ing proteins responsible. However, there are questions as to the stability of this procedure over multiple experiments.

Keith A. Baggerly; Jeffrey S. Morris; Kevin R. Coombes



Reproducibility of SELDI-TOF Protein Patterns in Serum: Comparing Data Sets from Dieren t Experiments  

Microsoft Academic Search

Motivation: There has been much interest in using pat- terns derived from surface-enhanced laser desorption and ionization (SELDI) protein mass spectra from serum to dieren tiate samples from patients both with and with- out disease. Such patterns have been used without iden- tication of the underlying proteins responsible. How- ever, there are questions as to the stability of this pro-

Keith A. Baggerly; Jerey S. Morris; Kevin R. Coombes


Serum extracellular vesicle protein levels are associated with acute coronary syndrome  

PubMed Central

Aims: Biomarkers are essential in the early detection of acute coronary syndromes (ACS). Serum extracellular vesicles are small vesicles in the plasma containing protein and RNA and have been shown to be involved in ACS-related processes like apoptosis and coagulation. Therefore, we hypothesized that serum extracellular vesicle protein levels are associated with ACS. Methods and results: Three serum extracellular vesicle proteins potentially associated with ACS were identified with differential Q-proteomics and were evaluated in 471 frozen serum samples of ACS-suspected patients presenting to the emergency department (30% of whom had an ACS). Protein levels were measured after vesicle isolation using ExoQuick. Mean serum extracellular vesicle concentration of the different proteins was compared between ACS and non-ACS patients. Selected proteins were tested in a univariate logistic regression model, as well as in a multivariate model to adjust for cardiovascular risk factors. A separate analysis was performed in men and women. In the multivariate logistic regression analysis, polygenic immunoglobulin receptor, (pIgR; OR 1.630, p=0.026), cystatin C (OR 1.641, p=0.021), and complement factor C5a (C5a, OR 1.495, p=0.025) were significantly associated with ACS, while total vesicle protein concentration was borderline significant. The association of the individual proteins with ACS was markedly stronger in men. Conclusions: These data show that serum extracellular vesicle pIgR, cystatin C, and C5a concentrations are independently associated with ACS and that there are pronounced gender differences. These observations should be validated in a large, prospective study to assess the potential role of vesicle content in the evaluation of patients suspected of having an ACS.

de Hoog, Vince C; Schoneveld, Arjan H; Wang, Jiong-Wei; van de Weg, Sander M; Sze, Siu Kwan; van Keulen, J Karlijn; Hoes, Arno W; den Ruijter, Hester M; de Kleijn, Dominique PV; Mosterd, Arend



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

PubMed Central

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

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



Serum interleukin 18 and interleukin 18 binding protein in rheumatoid arthritis  

Microsoft Academic Search

Objective: To measure serum interleukin 18 (IL18) and IL18 binding protein (IL18BP) levels in patients with inflammatory arthropathies, and to identify associations with disease status and the response to treatment.Methods: Serum samples were obtained before and after methotrexate treatment from patients with rheumatoid arthritis (RA) and psoriatic arthritis (PsA) attending an early arthritis clinic. IL18 and IL18BP were measured by

B Bresnihan; P Roux-Lombard; E Murphy; D Kane; O FitzGerald; J-M Dayer



Serum Deprivation and Protein Synthesis Inhibition Induce Two Different Apoptotic Processes in N18 Neuroblastoma Cells  

Microsoft Academic Search

N18 are murine neuroblastoma cells that underwent cell death upon serum deprivation or inhibition of protein synthesis by means of cycloheximide (CHX). In both cases, an ultrastructural morphology and an internucleosomal pattern of DNA fragmentation typical of apoptosis were found. However, electron microscopy revealed abundant lipid vesicles in the cytoplasm of CHX-treated cells that were not found in their serum-deprived

J. Boix; J. Fibla; V.-J. Yuste; J. M. Piulats; N. Llecha; J. X. Comella



Preanalytic Influence of Sample Handling on SELDI-TOF Serum Protein Profiles  

Microsoft Academic Search

Background: High-throughput proteomic methods for disease biomarker discovery in human serum are prom- ising, but concerns exist regarding reproducibility of results and variability introduced by sample handling. This study investigated the influence of different pre- analytic handling methods on surface-enhanced laser desorption\\/ionization time-of-flight mass spectrometry (SELDI-TOF MS) protein profiles of prefractionated serum. We investigated whether older collections with longer sample

John F. Timms; Elif Arslan-Low; Aleksandra Gentry-Maharaj; Zhiyuan Luo; Davy T'Jampens; Vladimir N. Podust; Jeremy Ford; Eric T. Fung; Alex Gammerman; Ian Jacobs; Usha Menon



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

PubMed Central

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

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



The vitamin E-binding protein afamin increases in maternal serum during pregnancy  

PubMed Central

Background Afamin is a liver-derived plasma glycoprotein with vitamin E-binding properties and a putative function in fertility. This study evaluated serum afamin concentrations during and postpartum to uncomplicated pregnancies and investigated a potential association between afamin concentrations and pregnancy outcome. Methods Afamin serum concentrations were measured in women with uncomplicated pregnancies in a retrospective cohort (n = 466) at different gestational ages and a prospective observational study (n = 76) in the first, second and third trimester. Furthermore, afamin was determined in the first trimester in a cross-sectional pilot study including women with preeclampsia (PE), pregnancy-induced hypertension (PIH) and women without pregnancy complications (n = 13 each). Finally, expression of afamin was investigated in human placental tissue by RT-PCR and immunohistochemistry. Results Afamin concentrations increased linearly almost two-fold during pregnancy in both retrospective and prospective studies in women without pregnancy complications with median afamin serum concentrations of 61.9 mg/l, 79.6 mg/l, and 98.6 mg/l in the first, second, and third trimester, respectively. After delivery, median afamin concentrations decreased to baseline values of 54.6 mg/l. In the pilot study with pregnancy complications, women with PE displayed significantly higher median afamin concentrations than did women with uncomplicated pregnancy (70.0 mg/l vs. 55.4 mg/l, P = 0.007). Expression analyses revealed no placental afamin expression at either mRNA or protein level in uncomplicated pregnancy. Conclusion A linear increase in the maternally expressed glycoprotein afamin during pregnancy may serve as basic reference for subsequent investigations of afamin in pregnancy-related disorders.

Hubalek, Michael; Buchner, Hannes; Mortl, Manfred G.; Schlembach, Dietmar; Huppertz, Berthold; Firulovic, Branka; Kohler, Wolfgang; Hafner, Erich; Dieplinger, Benjamin; Wildt, Ludwig; Dieplinger, Hans



Serum ?-Trace Protein and Risk of Mortality in Incident Hemodialysis Patients  

PubMed Central

Summary Background and objectives Residual kidney function in dialysis patients is associated with better survival, but there are no simple methods for its assessment. ?-Trace protein is a novel endogenous filtration marker of kidney function that is not removed during hemodialysis and may serve as a marker for residual kidney function similar to serum creatinine in patients not on dialysis. The objective of this study was to determine the association of serum ?-trace protein with mortality in incident hemodialysis patients. Design, setting, participants, & measurements Serum ?-trace protein was measured in baseline samples from 503 participants of a national prospective cohort study of incident dialysis patients with enrollment during 1995–1998 and follow-up until 2004. Outcomes were all-cause and cardiovascular disease mortality analyzed using Cox regression adjusted for demographic, clinical, and treatment factors. Results Serum ?-trace protein levels were higher in individuals with no urine output compared with individuals with urine output (9.0±3.5 versus 7.6±3.1 mg/L; P<0.001). There were 321 deaths (159 deaths from cardiovascular disease) during follow-up (median=3.3 years). Higher ?-trace protein levels were associated with higher risk of mortality. The adjusted hazard ratio and 95% confidence interval for all-cause mortality per doubling of serum ?-trace protein was 1.36 (1.09–1.69). The adjusted hazard ratios (95% confidence intervals) for all-cause mortality in the middle and highest tertiles compared with the lowest tertile were 0.95 (0.69–1.32) and 1.72 (1.25–2.37). Similar results were noted for cardiovascular disease mortality. Conclusions The serum level of ?-trace protein is an independent predictor of death and cardiovascular disease mortality in incident hemodialysis patients.

Parekh, Rulan S.; Jaar, Bernard G.; Plantinga, Laura C.; Oberai, Pooja C.; Eckfeldt, John H.; Levey, Andrew S.; Powe, Neil R.; Coresh, Josef



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.



The Content of Serum Protein in the Blood of Rainbow Trout (Soderzhanie Belka V Syvorotke Korovi Raduzhnoi Foreli).  

National Technical Information Service (NTIS)

The content of blood serum protein in rainbow trout changes in relation to age. With increased age the percent of blood protein increases. In two month old fingerlings the content of blood serum protein comprised 3.5%, at five months - 5.3, at 14 months- ...

N. V. Koroleva



An Investigation on the Blood-Serum Proteins of Chalcides ocellatus (Sauria: Scincidae) Populations from Southern Anatolia  

Microsoft Academic Search

In this study, blood-serum proteins of 22 adult (11 , 11 ) Chalcides ocellatus (ocelated skink) specimens collected from various localities in Southern Anatolia are investigated qualitatively and quantitatively by means of polyacrylamide disc electrophoresis and densitometry. The species' serum protein electropherograms and protein distribution patterns are established for the first time.

Hüseyin ARIKAN; Mehmet K. ATATÜR; Ahmet MERMER


Feasibility study of gel filtration as a separation method for blood serum proteins in combination with PIXE analysis.  


The combination between a protein separation technique and the PIXE method has a great potential for large surveys, including thousands of samples, in which multielemental analysis is required. Gel filtration with a Sephadex G-200 gel and a TRIS-acetate buffer was used for separating proteins in human serum. The fractions were doped with an yttrium/vanadium standard and then concentrated and pipeted onto Kimfol(™) backing foils. Using the PIXE technique, the distributions of Fe, Cu, Zn, and Se, with respect to molecular size, were found, indicating binding to specific proteins. Sulfur and phosphorus were found to correlate well with the protein content measured by UV-absorption at 280 nm. Further developments and tests on the protein separation technique is required, taking restrictions imposed by the PIXE method into consideration. PMID:24254619

Pallon, J; Pakarinen, P; Malmqvist, K; Roland Akselsson, K



Various protein and albumin corrections of the serum fructosamine concentration in the diagnosis of canine diabetes mellitus  

Microsoft Academic Search

Fructosamines are formed when glucose reacts non-enzymatically with amino groups on proteins, and previous studies have indicated that the serum fructosamine concentration could be of importance in the diagnosis of canine diabetes mellitus. Owing to the connection between the protein\\/albumin concentration and serum fructosamine concentration, it has been suggested that the serum fructosamine concentration should be corrected for the protein\\/albumin

A. L. Jensen



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.

Agoston, Denes V.; Elsayed, Mohammad



Effect of chromium chloride supplementation on glucose tolerance and serum lipids including high-density lipoprotein of adult men13  

Microsoft Academic Search

Chromium deficiency may cause insulin resistance, hyperinsulinemia, impaired glucose tolerance, and hyperlipidemia, reversed by chromium supplementation. The effect of chromium supplementation on serum lipids and glucose tolerance was tested in a double-blind 12- wk study of 23 healthy adult men aged 31 to 60 yr. Either 200 pg trivalent chromium in 5 ml water (Cr) or 5 ml plain water

Rebecca Riales; Margaret J. Aibrink


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.

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



Phenotype and allele frequencies of some serum protein polymorphisms in populations of the Balkans.  


Within a study of the genetics of Southeastern European populations seven serum protein polymorphisms (AMY2, BF, C3, CP, GC, HPA, TF) were examined in two samples of Aromuns and one reference sample (Musequiar-Aromuns from Dukasi in Albania, Moskopolian-Aromuns from Krusevo, Republic of Macedonia, and Macedonians from Skopje). The neighbor joining tree as well as the principal component analysis show results which do not correspond well to the geographic and historic background. This indicates that in the present case the serum protein polymorphisms give no clearly defined information about the relationships between the Balkan populations and to the origin of Aromuns. PMID:15648851

Scheil, H G; Schmidt, H D; Efremovska, L; Mikerezi, I; Huckenbeck, W



Differentially Expressed RNA from Public Microarray Data Identifies Serum Protein Biomarkers for Cross-Organ Transplant Rejection and Other Conditions  

Microsoft Academic Search

Serum proteins are routinely used to diagnose diseases, but are hard to find due to low sensitivity in screening the serum proteome. Public repositories of microarray data, such as the Gene Expression Omnibus (GEO), contain RNA expression profiles for more than 16,000 biological conditions, covering more than 30% of United States mortality. We hypothesized that genes coding for serum- and

Rong Chen; Tara K. Sigdel; Li Li; Neeraja Kambham; Joel T. Dudley; Szu-chuan Hsieh; R. Bryan Klassen; Amery Chen; Tuyen Caohuu; Alexander A. Morgan; Hannah A. Valantine; Kiran K. Khush; Minnie M. Sarwal; Atul J. Butte



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.

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



Serum C-reactive protein concentrations in early abdominal and pulmonary sepsis  

PubMed Central

Objectives To evaluate the C-reactive protein serum levels in patients with pulmonary and abdominal sepsis during the first five days of sepsis progression. Methods The present investigation was a retrospective cohort study conducted at the university hospital with 345 patients who were admitted to the intensive care unit and diagnosed with sepsis of pulmonary or abdominal origin. Serum C-reactive protein concentrations were measured by the turbidimetric immunoassay. For analysis of C-reactive protein, day 1 was defined as the day on which the patient was clinically diagnosed with sepsis. Results Thirty-four patients with sepsis (9.8%), 114 patients with severe sepsis (33.0%), and 197 patients with septic shock (57.2%) were evaluated. The age of the patients was 56.4±19.8 years. The serum C-reactive protein concentrations were higher on the day of sepsis diagnosis in the group with abdominal infection compared with the group with pulmonary sepsis (17.8±10.1 mg/dL versus 14.9±11.1 mg/dL, p=0.025) and remained significantly higher during the first five days of sepsis progression. Conclusion The serum C-reactive protein concentrations were significantly higher in the patients with abdominal sepsis compared with the patients with pulmonary sepsis during the first five days of sepsis progression.

Orati, Juliane Agustini; Almeida, Patricia; Santos, Vanessa; Ciorla, Gustavo; Lobo, Suzana Margareth



Serum Glial Fibrillary Acidic Protein as a Specific Marker for Necrotizing Meningoencephalitis in Pug Dogs  

PubMed Central

ABSTRACT To evaluate the ability of serum glial fibrillary acidic protein (GFAP) concentration as a diagnostic marker for canine central nervous system (CNS) disorders, sera from dogs with various CNS (n=47) and non-CNS (n=56) disorders were measured for GFAP by using an ELISA kit. Healthy Beagles (n=15) and Pug dogs (n=12) were also examined as controls. Interestingly, only Pug dogs with necrotizing meningoencephalitis (NME) showed elevated serum GFAP concentrations (<0.01 to 1.14 ng/ml), while other breeds of dogs with NME did not. Among the Pug dogs with NME, serum GFAP concentrations did not correlate with their clinical features, such as ages or survival times. Our data indicate the usefulness of serum GFAP as a novel marker for Pug dogs with NME.

MIYAKE, Hizuru; INOUE, Akiko; TANAKA, Miho; MATSUKI, Naoaki



Monocyte chemoattractant protein-1 serum levels in ovarian cancer patients  

Microsoft Academic Search

The chemokine monocyte chemoattractant protein (MCP)-1 is an important mediator of monocyte infiltration in various solid tumours of epithelial origin. The aim of the present study was to evaluate the role of MCP-1 in the natural history of ovarian cancer and to determine its value as differentiation marker and prognostic marker regarding disease free and overall survival. This retrospective study

L Hefler; C Tempfer; G Heinze; K Mayerhofer; G Breitenecker; S Leodolter; A Reinthaller; C Kainz



Serum antibodies against the heat shock protein 60 are elevated in patients with osteosarcoma.  


Osteosarcoma is the most frequent malignant bone tumor, mainly occurring in the second and third decade of life. Diagnosis is limited to clinical symptoms, radiology and histology, but so far no diagnostic laboratory tests are available. Heat shock proteins (hsp), highly conserved proteins performing vital intracellular chaperoning functions and preventing cells from death, have been shown to be involved in tumor immunity. We analyzed 75 sera from 23 patients with high-grade osteosarcoma, 8 patients with chondrosarcoma, 10 patients with Ewing's sarcoma, 5 patients with soft tissue sarcoma, 11 patients with benign bone tumors at the time of diagnosis and from 18 healthy controls with an indirect one-site enzyme linked immunosorbent assay (ELISA) for the presence of anti-hsp60 and 70 antibodies. In these assays 10/23 osteosarcoma patients (43%) had anti-hsp60 antibodies with a mean +/- S.D. titer of 0.382 +/- 0.243 U/ml. Only one of the 18 healthy controls (1/18, 5.6%; titer 0.22 U/ml), two of the Ewing's sarcoma patients (2/10, 20%; titer 0.2 +/- 0.09 U/ml), two of the patients with a benign bone tumor (2/11, 18%; titer 0.22 +/- 0.16 U/ml) and one of the chondrosarcoma patients (1/8, 12.5%; titer 0.14 U/ml) were positive, whereas all others, including all soft tissue sarcomas were negative throughout. Anti-hsp60 antibodies in patients with osteosarcoma are therefore significantly increased (p < 0.05). 19/23 (83%) of osteosarcoma biopsy specimens expressed hsp60 immunohistochemically and all specimens from patients with a positive anti-hsp60 serum titer expressed hsp60. The level of the anti-hsp60 antibodies did not correlate with clinical parameters such as response to preoperative chemotherapy, duration of symptoms, age, gender, tumor size, serum alkaline-phosphatase levels and metastases. Although no difference in anti-hsp70 antibodies could be observed between sera from patients and healthy controls, a positive correlation was found for the presence of anti-hsp70 serum antibodies and lung metastases at the time of diagnosis in osteosarcoma patients. These data suggest an increase of anti-hsp60 antibodies at the time of first diagnosis of osteosarcoma. These findings should therefore give rise to further investigations on a group of new markers for the diagnosis of osteosarcoma. PMID:10776793

Trieb, K; Gerth, R; Windhager, R; Grohs, J G; Holzer, G; Berger, P; Kotz, R



Serum concentration and interaction properties of MBL/ficolin associated protein-1.  


Recently, a novel protein named MBL/ficolin associated protein-1 (MAP-1) derived from the MASP1 gene through differential splicing was identified. In the present study, we established biochemical characteristics, determined the serum level and assessed the interactions between the lectin complement pathway (LCP) recognition molecules and MAP-1. We expressed recombinant MAP-1 in CHO DG44 cells, developed a quantitative ELISA assay based on a MAP-1 specific monoclonal capture antibody and measured the serum levels in 100 Danish blood donors. In addition we assessed the association properties between MAP-1 and Ficolin-2, -3 and MBL in serum using ELISA and density gradient ultra centrifugation. When recombinant MAP-1 was subjected to N-glycosidase F treatment the molecular mass decreased from ?45 kDa to ?40 kDa equivalent with the calculated molecular mass from the deduced amino acid sequence without the signal peptide. We found that serum MAP-1 was very stable when subjected to repeated freeze and thaw cycles. The mean serum concentration of MAP-1 was found to be 240 ng/ml (range: 115-466 ng/ml). MAP-1 was predominantly found in complex with Ficolin-3 and to a lesser degree with Ficolin-2 and MBL and by use of density gradient ultra centrifugation we could show that the major part of serum MAP-1 circulates in complex with the LCP molecules. In conclusion, these results show that MAP-1 is a highly stable glycosylated human serum protein found in complex with Ficolin-3, Ficolin-2 and MBL. PMID:21035894

Skjoedt, Mikkel-Ole; Hummelshoj, Tina; Palarasah, Yaseelan; Hein, Estrid; Munthe-Fog, Lea; Koch, Claus; Skjodt, Karsten; Garred, Peter



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


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

PubMed Central

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




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.



Changes in Specific Blood Serum Protein Levels Associated with Parturition in the Bovine1  

Microsoft Academic Search

Beginning about 14 weeks before parturition, the level of serum proteins in the blood of the bovine started increasing, to reach a maximum about four weeks before parturition; and then started decreasing, to reach a minimum at parturition. The drop at parturition was caused by a loss of immune B~- and ~-globulins and some a-globulins from the blood; the level

B. L. Larson; K. A. Kendall



Determination of serum proteins in the presence of dextran by means of the biuret reaction.  


264 biuret reagents for total protein determination in serum containing dextran were investigated. The usefulness of the method is dependent on proper ratio of the components of the reagent: concentrations of NaOH and copper sulphate, and the ratio of sodium-potassium tartrate to copper sulphate. PMID:557456

Gregor, A; Kostrzewska, E; Godorowska, W



Genetic Polymorphism of AHSG, FXIIIB, HP and PLG Serum Proteins in Six Jewish Groups in Israel  

Microsoft Academic Search

The allelic distribution of the polymorphic serum proteins AHSG, PLG, FXIIIB and HP was studied in six Jewish groups who migrated to Israel from the Middle East, North Africa, Rumania, Bulgaria, Central and Eastern Europe. The observed AHSG and PLG allele frequencies in these Jewish groups were more or less similar to the observed distributions in non-Jewish populations from their

S. Nevo; Margrit Oppong-Nketiah; M. Schröder; B. Cleve; A. J. Nabulsi; V. Chopra



Serum Proteins of Germ-Free Rats Fed Water-Soluble Diets.  

National Technical Information Service (NTIS)

The report presents data on serum proteins of germfree rats fed a chemically defined, water-soluble 'antigen free' diet which was filter-sterilized. This type of diet, composed almost entirely of low-molecularweight compounds and free of material of bacte...

B. S. Wostmann G. B. Olson J. R. Pleasants



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


Serum eosinophilic cationic protein may predict clinical course of wheezing in young children  

Microsoft Academic Search

Thirty eight children aged between 2 and 4 years with three or more episodes of wheezing were studied to evaluate the role of eosinophil inflammation and its relation to persistence of wheezing two years later. Serum eosinophilic cationic protein, total eosinophil count, total IgE, skin prick test, and clinical features were evaluated at visit 1. Two years later at a

J R Villa; G García; S Rueda; A Nogales



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



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, generate data sets that are comparable between experiments and laboratories, and have high throughput to support statistical studies. Here 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 now deglycosylated peptides by liquid chromatography electrospray ionization mass spectrometry, 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 with 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 J.; Aebersold, Reudi



Mechanisms of changes in composition of serum protein fractions after disturbance of the pancreatic exocrine function  

Microsoft Academic Search

Disturbance of the exocrine function of the pancreas in dogs with the pancreatic duct exteriorized may lead to substantial changes in the blood serum protein spectrum: well-marked dysproteinemia, a decrease in the albumin-globulin ratio, and the appearance of an additional globulin fraction between the \\/3- and ~2-globulins [ii, 12]. Intensive protein synthesis in the pancreas [14, 16, 19] evidently is

E. P. Fenina



Serum protein profiling and proteomics in autistic spectrum disorder using magnetic bead-assisted mass spectrometry  

Microsoft Academic Search

The pathophysiology of autistic spectrum disorder (ASD) is not fully understood and there are no diagnostic or predictive\\u000a biomarkers. Proteomic profiling has been used in the past for biomarker research in several non-psychiatric and psychiatric\\u000a disorders and could provide new insights, potentially presenting a useful tool for generating such biomarkers in autism. Serum\\u000a protein pre-fractionation with C8-magnetic beads and protein

Regina Taurines; Edward Dudley; Alexander C. Conner; Julia Grassl; Thomas Jans; Frank Guderian; Claudia Mehler-Wex; Andreas Warnke; Manfred Gerlach; Johannes Thome



Influence of serum proteins on erythrocyte uptake of digoxin and digitoxin  

Microsoft Academic Search

My purpose was to find out whether changes in serum protein content (albumin and gamma globulin) influenced erythrocyte uptake of glycosides and the capacity of erythrocytes to equilibrate glycoside levels when there are changes in protein binding. Heparinized human blood was incubated with 3H-digoxin, 6 ng\\/ml, and 3H-digitoxin, 44 ng\\/ml for 15 min. After centrifugation, distribution between plasma and erythrocytes

M L Gonçalves; Maria de Lurdes Gonçalves DrPharmSci



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

PubMed Central

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



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

PubMed Central

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



Seasonal and nutritional effects on serum proteins and urea concentration in the reindeer (Rangifer tarandus tarandus L.).  


1. The effects of seasonal conditions and nutrition on serum proteins and serum urea concentrations were studied in female reindeer and reindeer calves in Finland. With the exception of one group in winter, the reindeer were roaming wild in the forests. This one group was kept in captivity, out of doors, on a comparatively high nutritional plane. One group lived wild during the winter in very poor nutritional conditions. 2. A very clear seasonal variation in the serum protein and urea concentration was found. The serum protein concentration was low in late winter and increased rapidly during the summer, being high in the autumn. The serum urea concentration was also low in the winter and high in the summer. In the autumn, however, the serum urea concentration was again low. 3. Changes in the serum protein concentration were normally associated with the serum globulins. Only in the very poor-nutrition group did the albumin content decrease significantly. As a result of the large changes in the concentration of serum globulins, there were also considerable changes in the albumin: globulin ratio. 4. The serum protein concentration was much lower in the reindeer calves than in the adult reindeer. The concentration of globulins in particular was much lower than in the adults. PMID:1115752

Hyvärinen, H; Helle, T; Väyrynen, R; Väyrynen, P



Development of protein-deficient serum-free culture media for adherent cell lines suited for large scale cultivation  

Microsoft Academic Search

Summary Procedures are described for the devlopment of less expensive, protein-deficient serum-free media for the cultivation of mammalian adherent cell lines. Starting with a proteinrich chemically defined, serum-free culture medium, the concentrations of the expensive or highly concentrated proteins, or both, such as bovine serum albumin (BSA), insulin, or transferrin are systematically reduced one after the other until the minimal

Berthold G. D. Bödeker; Guy J. Berg; Ulrich Steiner; Guy Hewlett; H. Dieter Schlumberger



Serum and mucosal antibody responses to pneumococcal protein antigens in children: relationships with carriage status.  


Streptococcus pneumoniae causes significant morbidity and mortality especially in children. Some pneumococcal protein antigens can protect mice against infection. Little information is available concerning the nature of naturally acquired protective immunity to pneumococci in humans induced by these antigens. This study investigates the relationships between systemic and local antibody production and carriage in children. Children undergoing adenoidectomy (n=112, ages 2-12 years) were studied. Nasopharyngeal swabs were collected for pneumococcal culture. Serum and saliva were assayed for antibodies to several pneumococcal proteins: choline binding protein A (CbpA), pneumolysin (Ply), pneumococcal surface adhesin A (PsaA) and pneumococcal surface protein A (PspA). Adenoidal mononuclear cells (MNC) were cultured with pneumococcal culture supernatants or recombinant proteins. Cell culture supernatants were analyzed for antigen-specific antibodies. Carriage rates fell with age and serum levels of anti-CbpA, Ply and PspA rose. Anti-CbpA and -Ply serum and salivary IgG antibody levels were higher in children who were culture negative than those who were colonized. Antigen stimulation increased respective antigen-specific IgG production by adenoidal MNC and these responses were greater in those who were colonized than in culture-negative children. Antibodies to CbpA and Ply may protect children aged 2 years and older against pneumococcal colonization. Adenoids may be important local induction and effector sites for both mucosal and systemic antibody production to pneumococcal proteins in children. PMID:16342325

Zhang, Qibo; Bernatoniene, Jolanta; Bagrade, Linda; Pollard, Andrew J; Mitchell, Timothy J; Paton, James C; Finn, Adam



Serum heat shock protein-70 as a prognostic factor in patients with sudden sensorineural hearing loss  

Microsoft Academic Search

Objectives: In the cochlear cells, it has been known that HSP70 was expressed after ototoxic stimuli. We performed this study to investigate the increased serum HSP70 and its clinical role in the patients with sudden sensorineural hearing loss (SSNHL).Methods: A total of 67 patients with SSNHL and age-sex matched normal controls were included in this study. Their serum HSP70 levels

Sang-Won Yeo; Shi-nae Park; Kyoung-Ho Park



Serum Cytokine Profiles Associated with Specific Adjuvants Used in a DNA Prime-Protein Boost Vaccination Strategy  

PubMed Central

In recent years, heterologous prime-boost vaccines have been demonstrated to be an effective strategy for generating protective immunity, consisting of both humoral and cell-mediated immune responses against a variety of pathogens including HIV-1. Previous reports of preclinical and clinical studies have shown the enhanced immunogenicity of viral vector or DNA vaccination followed by heterologous protein boost, compared to using either prime or boost components alone. With such approaches, the selection of an adjuvant for inclusion in the protein boost component is expected to impact the immunogenicity and safety of a vaccine. In this study, we examined in a mouse model the serum cytokine and chemokine profiles for several candidate adjuvants: QS-21, Al(OH)3, monophosphoryl lipid A (MPLA) and ISCOMATRIX™ adjuvant, in the context of a previously tested pentavalent HIV-1 Env DNA prime-protein boost formulation, DP6-001. Our data revealed that the candidate adjuvants in the context of the DP6-001 formulation are characterized by unique serum cytokine and chemokine profiles. Such information will provide valuable guidance in the selection of an adjuvant for future AIDS vaccine development, with the ultimate goal of enhancing immunogenicity while minimizing reactogenicity associated with the use of an adjuvant. More significantly, results reported here will add to the knowledge on how to include an adjuvant in the context of a heterologous prime-protein boost vaccination strategy in general.

Buglione-Corbett, Rachel; Pouliot, Kimberly; Marty-Roix, Robyn; West, Kim; Wang, Shixia; Lien, Egil; Lu, Shan



2D Photonic Crystal Protein Hydrogel Coulometer for Sensing Serum Albumin Ligand Binding.  


Bovine and human serum albumin (BSA and HSA) are globular proteins that function as bloodstream carriers of hydrophobes such as fatty acids and drugs. We fabricated novel photonic crystal protein hydrogels by attaching 2D colloidal arrays onto pure BSA and HSA hydrogels. The wavelengths of the diffracted light sensitively report on the protein hydrogel surface area. The binding of charged species to the protein hydrogel gives rise to Donnan potentials that change the hydrogel volume causing shifts in the diffraction. These photonic crystal protein hydrogels act as sensitive Coulometers that monitor the hydrogel charge state. We find multiple high-affinity BSA and HSA binding sites for salicylate, ibuprofen and picosulfate by using these sensors to monitor binding of charged drugs. We demonstrate proof-of-concept for utilizing protein hydrogel sensors to monitor protein-ionic species binding. PMID:24766373

Cai, Zhongyu; Zhang, Jian-Tao; Xue, Fei; Hong, Zhenmin; Punihaole, David; Asher, Sanford A



Biophysical Insights into How Surfaces, Including Lipid Membranes, Modulate Protein Aggregation Related to Neurodegeneration  

PubMed Central

There are a vast number of neurodegenerative diseases, including Alzheimer’s disease (AD), Parkinson’s disease (PD), and Huntington’s disease (HD), associated with the rearrangement of specific proteins to non-native conformations that promotes aggregation and deposition within tissues and/or cellular compartments. These diseases are commonly classified as protein-misfolding or amyloid diseases. The interaction of these proteins with liquid/surface interfaces is a fundamental phenomenon with potential implications for protein-misfolding diseases. Kinetic and thermodynamic studies indicate that significant conformational changes can be induced in proteins encountering surfaces, which can play a critical role in nucleating aggregate formation or stabilizing specific aggregation states. Surfaces of particular interest in neurodegenerative diseases are cellular and subcellular membranes that are predominately comprised of lipid components. The two-dimensional liquid environments provided by lipid bilayers can profoundly alter protein structure and dynamics by both specific and non-specific interactions. Importantly for misfolding diseases, these bilayer properties can not only modulate protein conformation, but also exert influence on aggregation state. A detailed understanding of the influence of (sub)cellular surfaces in driving protein aggregation and/or stabilizing specific aggregate forms could provide new insights into toxic mechanisms associated with these diseases. Here, we review the influence of surfaces in driving and stabilizing protein aggregation with a specific emphasis on lipid membranes.

Burke, Kathleen A.; Yates, Elizabeth A.; Legleiter, Justin



The role of serum cholinesterase activity and S100B protein in the evaluation of organophosphate poisoning.  


The aim of this study was to investigate the role of serum cholinesterase (SChE) activity and S100B protein in the evaluation of patients with acute organophosphate (OP) poisoning. Patients with acute OP poisoning admitted to the emergency department were included in this cross-sectional study. Twenty healthy volunteers served as controls. The SChE activity and serum S100B were determined on admission. Patients were divided into two groups (low severity and high severity). Thirty-six patients diagnosed with acute OP poisoning were enrolled. Serum S100B concentrations were higher in patients than in the control group (p < 0.05). In the high-severity group, the SChE levels were lower and the S100Bs levels were higher than in the low-severity group. The SChE level was not different between survivors and nonsurvivors. S100B levels were higher in nonsurvivors than in survivors. According to receiver-operating characteristic curve analysis, the optimal cutoff value of serum S100B level to predict mortality was 236.5 pg/mL, with 71.4% sensitivity and 89.7% specificity. Our data suggest that initial SChE level is related to the clinical severity but not with mortality. S100B may be a useful marker in the assessment of clinical severity and prediction of mortality in acute OP poisoning. PMID:23424211

Yardan, T; Baydin, A; Acar, E; Ulger, F; Aygun, D; Duzgun, A; Nar, R



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:



MALDI/SELDI protein profiling of serum for the identification of cancer biomarkers.  


The ability to visualize the full depth of the serum proteome in a high-throughput manner is a major goal of clinical proteomics. Methodologies, which combine higher throughput with the ability to observe differential protein expression levels, have been applied to this goal. An example of such a system is the coupling of robotic sample processing to matrix-assisted laser desorption time of flight mass spectrometry (MALDI-TOF-MS). Within this paradigm is a modification of MALDI-TOF termed surface-enhanced laser desorption/ionization-TOF (SELDI-TOF). Both conventional MALDI and SELDI have been used to generate protein expression profiles reflective of potential peptide changes in serum. This information can be used to identify proteins, which may enable new diagnostic and therapeutic strategies. PMID:18287771

Cazares, Lisa H; Diaz, Jose I; Drake, Rick R; Semmes, O John



Quantitative analysis of microRNA in blood serum with protein-facilitated affinity capillary electrophoresis.  


MicroRNAs play an important role in gene regulation and disease etiology and are blood-based biomarkers of diseases. Here, we describe a protein-facilitated affinity capillary electrophoresis (ProFACE) method for ultra-sensitive direct miRNA detection as low as 300,000 molecules in 1 mL of blood serum, using single-stranded DNA binding protein (SSB) and double-stranded RNA binding protein (p19) as separation enhancers. This method utilizes either the selective binding of SSB to a fluorescent single-stranded DNA/RNA probe or the binding of p19 to miRNA-RNA probe duplex. PMID:24026701

Berezovski, Maxim V; Khan, Nasrin



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.



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



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


Distribution of the AB0 Blood Groups and the HP, TF, GC, PI and C3 Serum Proteins in Yakuts  

Microsoft Academic Search

In Yakut populations examined, polymorphisms of immunological and serum protein markers, including AB0 and Rhesus blood groups, HP, TF, GC, PI and C3, were revealed. Gene frequencies for the systems studied fell into the following ranges: AB0 system: r, 0.514 to 0.663; p, 0.136 to 0.306; q, 0.110 to 0.337; haptoglobin HP*1: 0.214 to 0.431; transferrin TF*C: 0.700 to 1.0;

L. A. Tarskaia; L. S. Bychkovskaia; G. V. Pai; S. B. Makarov; B. Pakendorf; V. A. Spitsyn



Comprehensive characterisation of pulmonary and serum surfactant protein D in COPD  

PubMed Central

Background Pulmonary surfactant protein D (SP-D) is considered as a candidate biomarker for the functional integrity of the lung and for disease progression, which can be detected in serum. The origin of SP-D in serum and how serum concentrations are related to pulmonary concentrations under inflammatory conditions is still unclear. Methods In a cross-sectional study comprising non-smokers (n = 10), young - (n = 10), elderly smokers (n = 20), and smokers with COPD (n = 20) we simultaneously analysed pulmonary and serum SP-D levels with regard to pulmonary function, exercise, repeatability and its quaternary structure by native gel electrophoresis. Statistical comparisons were conducted by ANOVA and post-hoc testing for multiple comparisons; repeatability was assessed by Bland-Altman analysis. Results In COPD, median (IQR) pulmonary SP-D levels were lower (129(68) ng/ml) compared to smokers (young: 299(190), elderly: 296(158) ng/ml; p < 0.01) and non-smokers (967(708) ng/ml; p < 0.001). The opposite was observed in serum, with higher concentrations in COPD (140(89) ng/ml) as compared to non-smokers (76(47) ng/ml; p < 0.01). SP-D levels were reproducible and correlated with the degree of airway obstruction in all smokers. In addition, smoking lead to disruption of the quaternary structure. Conclusions Pulmonary and serum SP-D levels are stable markers influenced by smoking and related to airflow obstruction and disease state. Smaller subunits of pulmonary SP-D and the rapid increase of serum SP-D levels in COPD due to exercise support the translocation hypothesis and its use as a COPD biomarker. Trial registration no interventional trial



Associations of Serum Skeletal Alkaline Phosphatase with Elevated C-Reactive Protein and Mortality  

PubMed Central

Summary Background and objectives Higher serum total alkaline phosphatase (AP) levels are associated with increased serum C-reactive protein (CRP) levels and mortality in the general and CKD populations. It is unclear to what extent these associations are related to bone disease. Design, setting, participants, & measurements In a nationally representative sample of 10,707 adult participants from the 1999–2004 National Health and Nutrition Examination Survey, serum nonskeletal AP levels were estimated from the measured serum skeletal and total AP levels. The associations of serum skeletal AP and nonskeletal AP levels with elevated serum CRP concentrations (>3 mg/L) and mortality were examined in multivariable models. Results Skeletal AP was not associated with elevated CRP (for each doubling in non-CKD: odds ratio [OR], 1.00; 95% confidence interval [95% CI], 0.90–1.11; in CKD: OR, 1.19; 95% CI, 0.83–1.70) or mortality (for each doubling in non-CKD: hazard ratio [HR], 1.10; 95% CI, 0.94–1.29; in CKD: HR, 0.98; 95% CI, 0.75–1.28). In contrast, nonskeletal AP was associated with elevated CRP (for each doubling in non-CKD: OR, 4.51; 95% CI, 3.80–5.35; in CKD: OR, 5.98; 95% CI, 3.40–10.51). Nonskeletal AP was associated with mortality in non-CKD (for each doubling: HR, 1.96; 95% CI, 1.37–2.80) but not in CKD (for each doubling: HR, 0.92; 95% CI, 0.51–1.67) (interaction P=0.03). Conclusions Bone disease is unlikely to account for the known associations of serum total AP with increased inflammation and mortality.

Filipowicz, Rebecca; Greene, Tom; Wei, Guo; Cheung, Alfred K.; Raphael, Kalani L.; Baird, Bradley C.



Characterization of granulations of calcium and apatite in serum as pleomorphic mineralo-protein complexes and as precursors of putative nanobacteria.  


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; Martel, Jan; Young, David; Young, Andrew; Hung, Chin-Ming; Young, Lena; Chao, Ying-Jie; Young, James; Wu, Cheng-Yeu



Peroxidation of proteins and lipids in suspensions of liposomes, in blood serum, and in mouse myeloma cells  

Microsoft Academic Search

There is growing evidence that proteins are early targets of reactive oxygen species, and that the altered proteins can in turn damage other biomolecules. In this study, we measured the effects of proteins on the oxidation of liposome phospholipid mem- branes, and the formation of protein hydroperoxides in serum and in cultured cells ex- posed to radiation-generated hydroxyl free radicals.

Janusz M. Gebicki; Juan Du; James Collins; Helen Tweeddale



Blood eosinophils and serum eosinophil cationic protein in patients with acute and chronic urticaria  

PubMed Central

We have analysed the relationship of blood eosinophil count and serum eosinophil cationic protein (ECP) levels in patients with acute and chronic idiopathic urticaria. The ECP levels and eosinophil counts were measured in the peripheral blood of 15 patients with acute urticaria, 25 with chronic idiopathic urticaria and 10 normal healthy subjects. Blood eosinophil counts and serum ECP levels increased in all patients with acute urticaria. Concerning patients affected by chronic urticaria, taking into account the recrudescence of the disease at the moment of taking the blood sample, only symptomatic patients showed increased eosinophil blood values whereas serum ECP levels were increased both in symptomatic and asymptomatic patients. Furthermore, serum ECP levels in chronic urticaria did not correlate with the peripheral eosinophil counts, as they did in acute urticaria. The results of the present study indicate that eosinophils may play a role in the inflammatory mechanisms in patients with acute and chronic urticaria showing a positive correlation between serum ECP levels and disease activity.

Lorenzo, G. Di; Mansueto, P.; Melluso, M.; Candore, G.; Cigna, D.; Pellitteri, M. E.; Salvo, A. Di



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.

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



Biomarkers of Exposure to Toxic Substances Volume 7: Identification of Potential Serum Protein Biomarkers Indicative of Low Level Kidney Degradation in Response to Toxin Exposures.  

National Technical Information Service (NTIS)

Potential serum biomarkers to subclinical nephrotoxin exposures were evaluated based on differential protein expression between control and dosed samples in rat serum. Proteins of interest demonstrated up-regulation at a minimum 1.5 fold increase in prote...

C. A. Mauzy C. L. Woolard P. A. Shiyanov



Identification and verification of heat shock protein 60 as a potential serum marker for colorectal cancer  

PubMed Central

Colorectal cancer (CRC) is a major public health issue worldwide, and novel tumor markers may contribute to its efficient management by helping in early detection, prognosis or surveillance of disease. The aim of our study was to identify new serum biomarkers for CRC, and we followed a phased biomarker discovery and validation process to obtain an accurate preliminary assessment of potential clinical utility. We compared colonic tumors and matched normal tissue from 15 CRC patients, using two-dimensional difference gel electrophoresis (2D-DIGE), and identified 17 proteins that had significant differential expression. These results were further confirmed by western blotting for heat shock protein (HSP) 60, glutathione-S-transferase Pi, ?-enolase, T-complex protein 1 subunit ?, and leukocyte elastase inhibitor, and by immunohistochemistry for HSP60. Using mAbs raised against HSP60, we developed a reliable (precision of 5–15%) and sensitive (0.3 ng·mL?1) immunoassay for the detection of HSP60 in serum. Elevated levels of HSP60 were found in serum from CRC patients in two independent cohorts; the receiver-operating characteristic curve obtained in 112 patients with CRC and 90 healthy controls had an area under the curve (AUC) of 0.70, which was identical to the AUC of carcinoembryonic antigen. Combination of serum markers improved clinical performance: the AUC of a three-marker logistic regression model combining HSP60, carcinoembryonic antigen and carbohydrate antigen 19-9 reached 0.77. Serum HSP60 appeared to be more specific for late-stage CRC; therefore, future studies should evaluate its utility for determining prognosis or monitoring therapy rather than early detection.

Hamelin, Celine; Cornut, Emilie; Poirier, Florence; Pons, Sylvie; Beaulieu, Corinne; Charrier, Jean-Philippe; Haidous, Hader; Cotte, Eddy; Lambert, Claude; Piard, Francoise; Ataman-Onal, Yasemin; Choquet-Kastylevsky, Genevieve



Serum Protein Profiling of Systemic Lupus Erythematosus and Systemic Sclerosis Using Recombinant Antibody Microarrays*  

PubMed Central

Systemic lupus erythematosus (SLE) and systemic sclerosis (SSc) are two severe autoimmune connective tissue diseases. The fundamental knowledge about their etiology is limited and the conditions display complex pathogenesis, multifaceted presentations, and unpredictable courses. Despite significant efforts, the lack of fully validated biomarkers enabling diagnosis, classification, and monitoring of disease activity represents significant unmet clinical needs. In this discovery study, we have for the first time used recombinant antibody microarrays for miniaturized, multiplexed serum protein profiling of SLE and SSc, targeting mainly immunoregulatory proteins. The data showed that several candidate SLE-associated multiplexed serum biomarker signatures were delineated, reflecting disease (diagnosis), disease severity (phenotypic subsets), and disease activity. Selected differentially expressed markers were validated using orthogonal assays and a second, independent patient cohort. Further, biomarker signatures differentiating SLE versus SSc were demonstrated, and the observed differences increased with severity of SLE. In contrast, the data showed that the serum profiles of SSc versus healthy controls were more similar. Hence, we have shown that affinity proteomics could be used to de-convolute crude, nonfractionated serum proteomes, extracting molecular portraits of SLE and SSc, further enhancing our fundamental understanding of these complex autoimmune conditions.

Carlsson, Anders; Wuttge, Dirk M.; Ingvarsson, Johan; Bengtsson, Anders A.; Sturfelt, Gunnar; Borrebaeck, Carl A. K.; Wingren, Christer



Smoking and lung cancer-induced changes in N-glycosylation of blood serum proteins  

PubMed Central

Glycosylation is a key post-translational protein modification which appears important in malignant transformation and tumor metastasis. Abnormal glycosylation of different proteins can often be measured in the blood serum. In this study, we extend our serum-based structural investigations to samples provided by patients diagnosed with lung cancer, paying particular attention to the effects of smoking on the serum glycomic traces. Following a battery of glycomic tests, we find that several fucosylated tetra-antennary structures with varying degrees of sialylation are increased in their abundances in control samples provided by the former smokers, with further elevations in the lung cancer patients who were former smokers. Further detailed investigations demonstrated that the level of outer-arm fucosylation was elevated in the control samples of the former smokers and again in the lung cancer samples provided by the former smokers. This trend was particularly noticeable for the tri- and tetra-antennary structures. Different ratios of sialylation linkages were also observed that could be correlated with the different states of health and smoking status. Decreases in the abundance levels of isomers with two and three ?2,3-linked sialic acids and an increased abundance of an isomer with two ?2,6-linked sialic acids were noted for a fucosylated tri-sialylated tri-antennary glycan. These results demonstrate the long-term effects of smoking on glycomic profiles and that this factor needs to be considered in these and other serum-based analyses.

Vasseur, Jacqueline A; Goetz, John A; Alley, William R; Novotny, Milos V



Characterization of the ovine complement 4 binding protein-beta (C4BPB) chain as a serum biomarker for enhanced diagnosis of sheep scab.  


Sheep scab, caused by the highly contagious mite Psoroptes ovis, is endemic in a number of sheep-producing countries worldwide, and is a major animal welfare and economic concern. Recent developments in the diagnosis of sheep scab include a highly sensitive and specific serum antibody-based assay which can be used to indicate exposure to the parasite but not necessarily current disease status. Here, a transcriptomic and bioinformatics analysis of the circulating leukocytes of sheep with active P. ovis infestation indicated that the transcription levels of complement 4 binding protein beta (C4BPB) increased by 12 fold from pre-infestation to 6 weeks post-infestation. Semi-quantitative studies confirmed increased serum C4BPB protein levels in sheep infested with P. ovis. To quantify this serum protein response and characterize ovine C4BPB as a biomarker for active P. ovis infestation, the ovine C4BPB gene was sequenced, a recombinant protein expressed, antibodies against this protein were raised in rabbits and a sandwich ELISA developed. The results from this assay indicated that serum C4BPB protein levels increased 4-fold from pre-infestation to 6 weeks post-infestation, which demonstrated the potential of the assay to quantify C4BPB in sheep sera and indicated the potential of C4BPB as a biomarker of current disease status in sheep post-infestation and post-treatment. PMID:23542335

Wells, Beth; Burgess, Stewart T G; Nisbet, Alasdair J



Evaluation of 14 commercial HIV1\\/HIV2 antibody assays using serum panels of different geographical origin and clinical stage including a unique seroconversion panel  

Microsoft Academic Search

The performance of 14 commercially available HIV-1\\/2 antibody assays were compared using well-characterized serum panels containing in total 1500–1800 sera. The panels included consecutive HIV-negative blood donor sera from Sweden, unselected blood donor and patient sera from Tanzania and unselected sera from outpatient clinics in Guinea-Bissau. Furthermore selected HIV-1 antibody positive sera from Sweden and Tanzania and HIV-2 antibody positive

Rigmor Thorstensson; Sören Andersson; Stefan Lindbäck; Francisco Dias; Fred Mhalu; Hans Gaines; Gunnel Biberfeld



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

PubMed Central

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

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



Postmortem serum protein S100B levels with regard to the cause of death involving brain damage in medicolegal autopsy cases  

Microsoft Academic Search

The protein S100 is an acidic calcium-binding protein, and the subunit S100B is the most abundantly found in the brain. The aim of the present study was a comprehensive analysis of serum S100B levels in medicolegal autopsy cases (within 48h postmortem, n=283) including victims with head and non-head injuries and also non-injury fatalities with regard to the cause of death

Dong-Ri Li; Bao-Li Zhu; Takaki Ishikawa; Dong Zhao; Tomomi Michiue; Hitoshi Maeda



Decrease of in vitro serum protein binding of salicylate in rheumatoid arthritis.  


Free and bound fractions of salicylates were separated by equilibrium dialysis and measured by spectrofluorimetry in 27 patients with rheumatoid arthritis and in 16 controls. The results showed that in patients with rheumatoid arthritis, the binding of salicylate to proteins decreased in an overproportional manner with the decrease of serum albumin concentrations. This phenomenon was linked with the severity of the inflammatory syndrome. The saturation binding capacity per unit of protein concentration was lower in the patients suffering from active forms of the condition, a finding which suggests that the changes observed are not due only to quantitative changes in the serum albumins. This study confirms the importance of determining free salicylate concentrations in the treatment of patients with inflammatory diseases. PMID:6745302

Netter, P; Monot, C; Stalars, M C; Mur, J M; Royer, R J; Faure, G; Pourel, J; Martin, J; Gaucher, A



Formation of long-lived reactive species of blood serum proteins by the action of heat.  


It has been previously established that heat induces the formation of reactive oxygen species (ROS) in aqueous solutions. In biological systems, ROS cause oxidative damage predominantly to proteins due to their abundance and sensitivity to oxidation. Proteins oxidized by the action of X-rays represent long-lived reactive species, which trigger the secondary generation of ROS (Bruskov et al. (2012) [25]). Here we studied the possibility of formation of long-lived species of the blood serum proteins bovine serum albumin and bovine gamma-globulin in air-saturated solutions under the action of heat. It is shown that heat induces the generation of long-lived protein species, which in turn generate ROS ((1)?2, (·)O2(-), (·)O?, and H2O2). The formation of the long-lived reactive species of BSA and BGG with a half-life of about 4h induced by moderate hyperthermia was revealed using the chemiluminescence of protein solutions. It was found that long-lived reactive species of BSA and BGG cause prolonged generation of H2O2. The results obtained suggest that H2O2 produced by proteins after heating represents a messenger in signaling pathways and produces therapeutic effects in living organisms. PMID:24361896

Bruskov, Vadim I; Popova, Nelly R; Ivanov, Vladimir E; Karp, Olga E; Chernikov, Anatoly V; Gudkov, Sergey V



Serum acute phase protein concentrations in dogs with spirocercosis and their association with esophageal neoplasia - A prospective cohort study.  


Spirocerca lupi, the dog esophageal worm, typically induces formation of esophageal nodules, which may transform to sarcoma. Ante mortem discrimination between benign and malignant esophageal masses is challenging. Serum acute phase proteins (APPs) are utilized in diagnosis and prognosis of various canine diseases as markers of inflammation. This study characterized serum APPs concentrations in dogs with benign and malignant esophageal spirocercosis and evaluated their accuracy in differentiating benign from malignant lesions. Seventy-eight client-owned dogs with esophageal spirocercosis were included. Serum C-reactive protein (CRP), haptoglobin, serum-amyloid A (SAA) and albumin concentrations were measured upon diagnosis and follow-up visits, and compared with healthy dogs, and between malignant and benign cases. Haptoglobin, CRP and SAA concentrations were higher, and albumin concentration was lower (P<0.001 for all) in infected dogs compared to healthy controls. Dogs with suspected neoplasia had significantly higher CRP (P=0.011), haptoglobin (P=0.008) and SAA (P=0.05), and lower albumin (P=0.012) concentrations compared to dogs with benign esophageal nodules. APPs moderately discriminated between suspected malignant and benign esophageal disease. None of the dogs with suspected neoplasia had concurrent normal concentrations of all APPs. The present results indicate that canine spirocercosis is characterized by an acute phase reaction, both at presentation and during treatment. When concentrations of all four APPs are within reference range, esophageal malignancy is highly unlikely. Although concentrations of all positive APPs were significantly higher in suspected neoplastic cases compared to benign ones, moderate discriminatory power limits their clinical use. Neither APP was useful to monitor response to treatment. PMID:24656552

Nivy, Ran; Caldin, Marco; Lavy, Eran; Shaabon, Keren; Segev, Gilad; Aroch, Itamar



Functional analysis of the larval serum protein gene promoter from silkworm Bombyx mori  

Microsoft Academic Search

The regulation region of larval serum protein gene, Bombyx mori. (BmLSP), consisting of the first intron, the first exon,\\u000a the central promoter region and 5’-upstream region, is cloned from genomic DNA from the silkworm variety of Suju × Minghu.\\u000a Using PCR and restriction endonuclease methods, a series of luciferase reporter plasmids, driven by different length of BmLSP\\u000a promoters, are constructed.

Shimming Tang; Yongzhu Yi; Xingjia Shen; Zhifang Zhang; Yiren Li; Jialu He



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



Serum Proteins of Germ-free Rats fed Water-soluble Diets  

Microsoft Academic Search

THE germ-free animal with its more controlled environment and reduced reticuloendothelial system has become a valuable tool in microbiological and immunological investigations. In the germ-free rat a deficit in immune proteins causes low levels of serum beta- and gamma-globulins1. However, the animal is still subject to the variable reticuloendothelial system stimulating factors occurring in the steam-sterilized diets generally used. This

B. S. Wostmann; G. B. Olson; J. R. Pleasants



Serum amyloid A: An acute-phase protein involved in tumour pathogenesis  

Microsoft Academic Search

.  The synthesis of acute-phase protein serum amyloid A (SAA) is largely regulated by inflammation- associated cytokines and\\u000a a high concentration of circulating SAA may represent an ideal marker for acute and chronic inflammatory diseases. However,\\u000a SAA is also synthesized in extrahepatic tissues, e.g. human carcinoma metastases and cancer cell lines. An increasing body\\u000a of in vitro data supports the concept

E. Malle; S. Sodin-Semrl; A. Kovacevic



A study on tuberculosis in buffaloes: some epidemiological aspects, along with haematological and serum protein changes  

Microsoft Academic Search

The study was conducted to ascertain the epidemiology, together with effects of bovine tuberculosis, on certain haematological parameters and serum proteins at two Livestock Experiment Stations in Pakistan. The results on prevalence of tuberculosis in buffaloes on the basis of comparative intradermal tuberculin test revealed it to be from as high as 8.48% (14\\/165) to as low as 2.45% (4\\/163)

Muhammad Tariq Javed; Mahmood Usman; Muhammad Irfan; Monica Cagiola



The serum bone morphogenetic protein-2 level in non-small-cell lung cancer patients  

Microsoft Academic Search

Bone morphogenetic proteins (BMPs) are members of the TGF-? superfamily, and some studies demonstrated that BMPs enhance tumorigenesis\\u000a and metastasis. The purpose of this study was to identify whether BMP-2 levels are elevated in the serum of non-small-cell\\u000a lung cancer (NSCLC) patients and are associated with the stage and overall survival. Blood samples from 150 patients with\\u000a NSCLC were analyzed.

Yoon Ji Choi; Seung Tae Kim; Kyong Hwa Park; Sang Cheul Oh; Jae Hong Seo; Sang Won Shin; Jun Suk Kim; Yeul Hong Kim


Effect of serum proteins on polystyrene nanoparticle uptake and intracellular trafficking in endothelial cells  

Microsoft Academic Search

The physico-chemical properties of nanoparticles (NPs), such as small dimensions, surface charge and surface functionalization,\\u000a control their capability to interact with cells and, in particular, with sub-cellular components. This interaction can be\\u000a also influenced by the adsorption of molecules present in biological fluids, like blood, on NP surface. Here, we analysed\\u000a the effect of serum proteins on 49 and 100 nm

Daniela Guarnieri; Angela Guaccio; Sabato Fusco; Paolo A. Netti


Serum intact molecule of bone Gla-protein in patients with abnormal bone and calcium metabolism  

Microsoft Academic Search

A circulating level of bone Gla-protein (BGP) has been estimated one of the most promising markers for bone turnover in patients\\u000a with metabolic bone diseases. Although previous bone histomorphometric studies have revealed that serum BGP levels are mainly\\u000a related to osteoblastic bone formation, multiple immunoreactive forms of BGP present in uremic sera are attributed to both\\u000a osteoclastic bone resorption and

Kiyoshi Nakatsuka; Takami Miki; Shigeichi Shoji; Yoshiki Nishizawa; Hirotoshi Morii



Serum protein profiling using surfactant-aided matrix-assisted laser desorption\\/ionization mass spectrometry  

Microsoft Academic Search

It has been shown previously that sodium dodecyl sulfate (SDS) can be used at its critical micellar concentration to increase the number of peptides detected in a peptide mixture by matrix-assisted laser desorption\\/ionization mass spectrometry (MALDI-MS). Here we demonstrate that, in a similar fashion, preparation of peptides and low molecular weight proteins from serum using SDS micelles improves the information

Rama Tummala; Patrick A. Limbach



Serum Resistance-Associated Protein Blocks Lysosomal Targeting of Trypanosome Lytic Factor in Trypanosoma brucei  

Microsoft Academic Search

Trypanosoma brucei brucei is the causative agent of nagana in cattle and can infect a wide range of mammals but is unable to infect humans because it is susceptible to the innate cytotoxic activity of normal human serum. A minor subfraction of human high-density lipoprotein (HDL) containing apolipoprotein A-I (apoA-I), apo- lipoprotein L-I (apoL-I), and haptoglobin-related protein (Hpr) provides this

Monika W. Oli; Laura F. Cotlin; April M. Shiflett; Stephen L. Hajduk



Producing High-Accuracy Lattice Models from Protein Atomic Coordinates Including Side Chains  

PubMed Central

Lattice models are a common abstraction used in the study of protein structure, folding, and refinement. They are advantageous because the discretisation of space can make extensive protein evaluations computationally feasible. Various approaches to the protein chain lattice fitting problem have been suggested but only a single backbone-only tool is available currently. We introduce LatFit, a new tool to produce high-accuracy lattice protein models. It generates both backbone-only and backbone-side-chain models in any user defined lattice. LatFit implements a new distance RMSD-optimisation fitting procedure in addition to the known coordinate RMSD method. We tested LatFit's accuracy and speed using a large nonredundant set of high resolution proteins (SCOP database) on three commonly used lattices: 3D cubic, face-centred cubic, and knight's walk. Fitting speed compared favourably to other methods and both backbone-only and backbone-side-chain models show low deviation from the original data (~1.5?Ĺ RMSD in the FCC lattice). To our knowledge this represents the first comprehensive study of lattice quality for on-lattice protein models including side chains while LatFit is the only available tool for such models.

Mann, Martin; Saunders, Rhodri; Smith, Cameron; Backofen, Rolf; Deane, Charlotte M.



Including explicit water molecules as part of the protein structure in MM/PBSA calculations.  


Water is the natural medium of molecules in the cell and plays an important role in protein structure, function and interaction with small molecule ligands. However, the widely used molecular mechanics Poisson-Boltzmann surface area (MM/PBSA) method for binding energy calculation does not explicitly take account of water molecules that mediate key protein-ligand interactions. We have developed a protocol to include water molecules that mediate ligand-protein interactions as part of the protein structure in calculation of MM/PBSA binding energies (a method we refer to as water-MM/PBSA) for a series of JNK3 kinase inhibitors. Improved correlation between water-MM/PBSA binding energies and experimental IC50 values was obtained compared to that obtained from classical MM/PBSA binding energy. This improved correlation was further validated using sets of neuraminidase and avidin inhibitors. The observed improvement, however, appears to be limited to systems in which there are water-mediated ligand-protein hydrogen bond interactions. We conclude that the water-MM/PBSA method performs better than classical MM/PBSA in predicting binding affinities when water molecules play a direct role in mediating ligand-protein hydrogen bond interactions. PMID:24432790

Zhu, Yong-Liang; Beroza, Paul; Artis, Dean R



Plant Protein Intake Is Associated with Fibroblast Growth Factor 23 and Serum Bicarbonate in Patients with CKD: The Chronic Renal Insufficiency Cohort Study  

PubMed Central

Background Protein from plant, as opposed to animal, sources may be preferred in chronic kidney disease (CKD), due to lower bioavailability of phosphate and lower nonvolatile acid load. Study Design Observational cross-sectional study. Setting & Participants 2938 participants with chronic kidney disease and information on dietary intake at the baseline visit in the Chronic Renal Insufficiency Cohort Study. Predictors Percentage of total protein from plant sources (% plant protein) was determined by scoring individual food items from the National Cancer Institute Diet History Questionnaire (DHQ). Outcomes Metabolic parameters, including serum phosphate, bicarbonate (HCO3), potassium, and albumin, plasma fibroblast growth factor 23 (FGF23), and parathyroid hormone (PTH), and hemoglobin. Measurements We modeled the association between % plant protein and metabolic parameters using linear regression. Models were adjusted for age, sex, race, diabetes, body mass index, eGFR, income, smoking, total energy intake, total protein intake, 24 hour urinary sodium, use of angiotensin converting enzyme inhibitors/angiotensin receptor blockers and use of diuretics. Results Higher % plant protein was associated with lower FGF23 (p=0.05) and higher HCO3 (p=0.01), but not with serum phosphate or PTH (p=0.9 and 0.5, respectively). Higher % plant protein was not associated with higher serum potassium (p=0.2), lower serum albumin (p=0.2) or lower hemoglobin (p=0.3). The associations of % plant protein with FGF23 and HCO3 did not differ by diabetes status, sex, race, CKD stage (2/3 vs. 4/5) or total protein intake (? 0.8 g/kg/d vs. >0.8 g/kg/d) (p-interaction > 0.10 for each). Limitations Cross-sectional study; Determination of % plant protein using the DHQ has not been validated. Conclusions Consumption of a higher percentage of protein from plant sources may lower FGF23 and raise HCO3 in patients with CKD.

Scialla, Julia J.; Appel, Lawrence J; Wolf, Myles; Yang, Wei; Zhang, Xiaoming; Sozio, Stephen M.; Miller, Edgar R.; Bazzano, Lydia A.; Cuevas, Magdalena; Glenn, Melanie J.; Lustigova, Eva; Kallem, Radhakrishna R.; Porter, Anna C.; Townsend, Raymond R.; Weir, Matthew R.; Anderson, Cheryl A.M.



A comparison of Tris-glycine and Tris-tricine buffers for the electrophoretic separation of major serum proteins.  


This paper compares different buffer systems for the electrophoretic separation of the five most abundant serum proteins on native-PAGE gel and cellulose membranes. A modified Tris-tricine system was shown to be superior for the separation of these serum proteins in a 7% m/v native-PAGE gel as compared with the traditionally used Tris-glycine and Tris-tricine methods. This modified Tris-tricine buffer system was also employed for the separation of serum proteins using a cellulose acetate membrane and very effective separation was observed as compared with the traditionally used Tris-barbital and Tris-glycine buffer systems. PMID:21818850

Haider, Syed R; Sharp, Barry L; Reid, Helen J



Quantitative analysis of microRNA in blood serum with protein-facilitated affinity capillary electrophoresis.  


MicroRNAs (miRNAs) are small (?22 nt) regulatory RNAs that are frequently deregulated in cancer and have shown promise as tissue- and blood-based biomarkers for cancer classification and prognostication. Here we present a protein-facilitated affinity capillary electrophoresis (ProFACE) assay for rapid quantification of miRNA levels in blood serum using single-stranded DNA binding protein (SSB) and double-stranded RNA binding protein (p19) as separation enhancers. The method utilizes either the selective binding of SSB to a single-stranded DNA/RNA probe or the binding of p19 to miRNA-RNA probe duplex. For the detection of ultralow amounts of miRNA without polymerase chain reaction (PCR) amplification in blood samples we apply off-line preconcentration of synthetic miRNA-122 from serum by p19-coated magnetic beads followed by online sample stacking in the ProFACE assay. The detection limit is 0.5 fM or 30?000 miRNA molecules in 1 mL of serum as a potential source of nai?ve miRNAs. PMID:21714529

Khan, Nasrin; Cheng, Jenny; Pezacki, John Paul; Berezovski, Maxim V



Shortcomings in methodology complicate measurements of serum retinol binding protein (RBP4) in insulin-resistant human subjects  

Microsoft Academic Search

Aims\\/hypothesis  Levels of retinol binding protein (RBP4) are increased in the serum of insulin-resistant human subjects even before overt\\u000a diabetes develops. RBP4 levels correlate with insulin resistance, BMI, WHR, dyslipidaemia and hypertension. Improvement of\\u000a insulin sensitivity with exercise training is associated with reduction in serum RBP4 levels. Therefore serum RBP4 may be\\u000a useful for early diagnosis of insulin resistance and for

T. E. Graham; C. J. Wason; M. Blüher; B. B. Kahn



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



Identification of Treatment Efficacy-Related Host Factors in Chronic Hepatitis C by ProteinChip Serum Analysis  

PubMed Central

Recent development of proteomic array technology, including protein profiling coupling ProteinChip array with surface-enhanced laser desorption ionization time-of-flight mass spectrometry (SELDI-TOF/MS), provides a potentially powerful tool for discovery of new biomarkers by comparison of its profiles according to patient phenotypes. We used this approach to identify the host factors associated with treatment response in patients with chronic hepatitis C (CHC) receiving a 48-wk course of pegylated interferon (PEG-IFN) alpha 2b plus ribavirin (RBV). Protein profiles of pretreatment serum samples from 32 patients with genotype 1b and high viral load were conducted by SELDI-TOF/MS by using the three different ProteinChip arrays (CM10, Q10, IMAC30). Proteins showed significantly different peak intensities between sustained virological responders (SVRs), and non-SVRs were identified by chromatography, SDS-PAGE, TOF/MS and tandem mass spectrometry (MS/MS) assay. Eleven peak intensities were significantly different between SVRs and non-SVRs. The three SVR-increased peaks could be identified as two apolipoprotein (Apo) fragments and albumin and, among the eight non–SVR-increased proteins, four peaks identified as two iron-related and two fibrogenesis-related protein fragments, respectively. Multivariate analysis showed that the serum ferritin and three peak intensity values (Apo A1, hemopexin and transferrin) were independent variables associated with SVRs, and the area under the receiver operating characteristic (ROC) curves for SVR prediction by using the Apo A1/hemopexin and hemopexin/transferrin were 0.964 and 0.936. In conclusion, pretreatment serum protein profiling by SELDI-TOF/MS is variable for identification of response-related host factors, which are useful for treatment efficacy prediction in CHC receiving PEG-IFN plus RBV. Our data also may help us understand the mechanism for treatment resistance and development of more effective antiviral therapy targeted toward the modulation of lipogenesis or iron homeostasis in CHC patients.

Fujita, Naoki; Nakanishi, Mamoru; Mukai, Jun; Naito, Yuuji; Ichida, Takafumi; Kaito, Masahiko; Yoshikawa, Toshikazu; Takei, Yoshiyuki



Early decrease of serum Clara cell protein in silica-exposed workers.  


Clara cell protein (CC16) is a 16 kDa protein secreted by nonciliated cells of the tracheobronchial tree; it has recently been proposed as a peripheral marker of respiratory epithelial injury. The concentration of CC16 was measured in the serum and, when available, in the sputum of 86 miners exposed to silica and of 86 control subjects matched for age, body mass index and smoking status (26 lifelong nonsmokers and 60 current smokers in both groups). Workers were exposed to silica-rich dust in a quarry for 15.2 months on average. No difference between exposed and control workers could be detected with regard to respiratory symptoms, chest radiographs or lung function tests. By contrast, the concentration of CC16 in serum was decreased in silica-exposed workers (geometric mean 12.3 micrograms.l-1) compared to controls (16.3 micrograms.l-1). The decrease was found both in lifelong nonsmokers (14.7 vs 21.9) and current smokers (11.3 vs 14.5). In the latter, tobacco smoking caused a decrease of serum CC16 that was additional to that associated with silica exposure. The determination of CC16 in sputum samples, judged to be reliable on the basis of the CC16/alpha-amylase concentration ratio (mostly from smokers), also revealed a reduction of CC16 following silica exposure (46.2 vs 106 mg.l-1). We conclude that alterations in the serum concentrations of CC16 probably reflect very early toxic effects of silica particles on the respiratory epithelium. This reinforces the view that serum CC16 is a sensitive marker, which might improve our ability to detect exposure to chemicals potentially harmful to the respiratory tract. PMID:7875262

Bernard, A M; Gonzalez-Lorenzo, J M; Siles, E; Trujillano, G; Lauwerys, R



Serum heat shock protein 47 levels are elevated in acute exacerbation of idiopathic pulmonary fibrosis.  


Little is known about the pathophysiology of acute exacerbation (AE) of idiopathic pulmonary fibrosis (IPF). Heat shock protein 47 (HSP47), a collagen-specific molecular chaperone, is essential for biosynthesis and secretion of collagen molecules. Previous studies in experimental animal fibrosis models have shown that downregulation of HSP47 expression reduces collagen production and diminishes fibrosis progression. In this study, serum HSP47 levels were evaluated to elucidate pathogenic differences involving HSP47 between AE-IPF and stable (S)-IPF. Subjects comprised 20 AE-IPF and 33 S-IPF patients. Serum levels of HSP47, Krebs von den Lungen-6 (KL-6), surfactant protein (SP)-A, SP-D, and lactate dehydrogenase (LDH) were measured. Immunohistochemical analysis of lung HSP47 expression was determined in biopsy and autopsy tissues diagnosed as diffuse alveolar damage (DAD) and usual interstitial pneumonia (UIP). Serum levels of HSP47 were significantly higher in AE-IPF than in S-IPF patients, whereas serum levels of KL-6, SP-A, and SP-D did not differ significantly. Receiver operating characteristic curves revealed that HSP47 was superior for discriminating AE-IPF and S-IPF. The cutoff for HSP47 resulting in the highest diagnostic accuracy was 559.4 pg/mL; sensitivity, specificity, and diagnostic accuracy were 100.0%, 93.9%, and 96.2%, respectively. Immunohistochemical analysis revealed that pulmonary HSP47 expression was greater in DAD than UIP tissues. Serum HSP47 was significantly higher in AE-IPF than in S-IPF patients, suggesting that underlying fibrogenic mechanisms involving HSP47 differ in the two conditions. PMID:23435730

Kakugawa, Tomoyuki; Yokota, Shin-Ichi; Ishimatsu, Yuji; Hayashi, Tomayoshi; Nakashima, Shota; Hara, Shintaro; Sakamoto, Noriho; Kubota, Hiroshi; Mine, Mariko; Matsuoka, Yasuhiro; Mukae, Hiroshi; Nagata, Kazuhiro; Kohno, Shigeru



Elevated Serum C-Reactive Protein as a Prognostic Marker in Small Cell Lung Cancer  

PubMed Central

Purpose Elevated C-reactive protein (CRP) is associated with poor prognosis in several tumor types. The purpose of this study was to investigate serum CRP as a prognostic marker in small cell lung cancer (SCLC). Materials and Methods The pretreatment serum CRP level was measured in 157 newly diagnosed SCLC patients, and correlation between serum CRP level and other clinical parameters was analyzed. Multivariate analyses were performed to find prognostic markers using Cox's proportional hazards model. Results The initial CRP concentration was within the normal range in 72 (45.9%) patients and elevated in 85 (54.1%) patients. There was a significant correlation between serum CRP level and the extent of disease (p<0.001), weight loss (p=0.029) and chest radiation (p=0.001). Median overall survival (OS) in the normal CRp group was significantly longer than with the high CRp group (22.5 months vs. 11.2 months, p<0.001). Extent of disease (p<0.001), age (p=0.025), and performance status (p<0.001) were additional prognostic factors on univariate analysis. On multivariate analysis, elevated serum CRp level was an independent prognostic factor for poor survival (HR=1.8; p=0.014), regardless of the extent of disease (HR=3.7; p<0.001) and performance status (HR=2.2; p<0.001). Conclusion High level of CRP was an independent poor prognostic serum marker in addition to previously well-known prognosticators in patients with SCLC.

Hong, Soojung; Kang, Young Ae; Cho, Byoung Chul



Serum proteins and enzymes in pregnant rats treated with epoxy resin or triethylenetetraamine stabilizer  

Microsoft Academic Search

A study was made of the serum and amniotic fluid levels of several acute phase reactants (seromucoid, haptoglobin, and sialic acid) and enzymatic activities (lactate dehydrogenase, leucylnaphthylamidase, ?-glutamyltranspeptidase, aminotransferases, cholmesterase, and alkaline and acid phosphatases) in pregnant and nonpregnant rats treated with epoxy resin Epidian 5 or triethylenetetraamine stabilizer. Changes produced by the action of the chemicals were similar, including

W. Dobryszycka; M. Warwas; A. Woytofi; J. Woyto?; J. Szacki



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


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

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



Proteomic study of serum proteins in a type 2 diabetes mellitus rat model by Chinese traditional medicine Tianqi Jiangtang Capsule administration.  


Proteomics technology was for the first time applied to investigate the changes of serum proteins levels in type 2 diabetes mellitus (T2DM) rat model after treated by Chinese traditional medicine Tianqi Jiangtang Capsule (ten normal Wistar rats, ten with T2DM and ten with T2DM administrated by Tianqi Jiangtang Capsule). In addition to two-dimensional polyacrylamide gel electrophoresis (2-DE), serum protein profiling in the three groups was further performed using matrix-assisted laser desorption/ionization mass spectrometry (MALDI-TOF-TOF/MS). 11 visualized spots were differentially regulated and identified as diabetes-associated proteins. All the samples in three groups were then analyzed by ELISA and estimated the 7 proteins which were found to vary. The distinct effect of T2DM induction on the pattern of rat serum includes the down-regulation of Apolipoprotein E, Apolipoprotein A-I, Ig gamma-2A chain C region, and up-regulation of Transthyretin (TTR), Haptoglobin (Hp), Serum amyloid P-componen (SAP), Prothrombin. The majority of those protein levels were interestingly restored to those of healthy rats after Tianqi Jiangtang Capsule treatment. PMID:20674218

Zhang, Shu-Xiang; Sun, Hui; Sun, Wen-Jun; Jiao, Guo-Zheng; Wang, Xi-Jun



In vivo Effects of Inhibitors of Protein Synthesis on Infection-Related Changes in Serum Amino Acids.  

National Technical Information Service (NTIS)

In rats exposed to Diplococcus pneumoniae, the infectious process is characterized by a decrease in serum amino acids, flux of amino acids from skeletal muscle to liver, and a subsequent increased synthesis of serum proteins. This process is mediated by a...

R. W. Wannemacher M. C. Powamda



Depletion of high-abundance proteins from serum by immunoaffinity chromatography: A MALDI-FT-MS study  

Microsoft Academic Search

Immunodepletion of high-abundance proteins from serum is a widely used initial step in biomarker discovery studies. In the present work we have investigated the reproducibility of the depletion step by comparing 250 serum samples from prostate cancer patients. All samples were depleted on a single immunoaffinity column over a time period of 6 weeks with automated peak detection and fraction

Lennard J. Dekker; Jan Bosman; Peter C. Burgers; Angelique van Rijswijk; Robert Freije; Theo Luider; Rainer Bischoff



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


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

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



Nephrin Regulates Lamellipodia Formation by Assembling a Protein Complex That Includes Ship2, Filamin and Lamellipodin  

PubMed Central

Actin dynamics has emerged at the forefront of podocyte biology. Slit diaphragm junctional adhesion protein Nephrin is necessary for development of the podocyte morphology and transduces phosphorylation-dependent signals that regulate cytoskeletal dynamics. The present study extends our understanding of Nephrin function by showing in cultured podocytes that Nephrin activation induced actin dynamics is necessary for lamellipodia formation. Upon activation Nephrin recruits and regulates a protein complex that includes Ship2 (SH2 domain containing 5? inositol phosphatase), Filamin and Lamellipodin, proteins important in regulation of actin and focal adhesion dynamics, as well as lamellipodia formation. Using the previously described CD16-Nephrin clustering system, Nephrin ligation or activation resulted in phosphorylation of the actin crosslinking protein Filamin in a p21 activated kinase dependent manner. Nephrin activation in cell culture results in formation of lamellipodia, a process that requires specialized actin dynamics at the leading edge of the cell along with focal adhesion turnover. In the CD16-Nephrin clustering model, Nephrin ligation resulted in abnormal morphology of actin tails in human podocytes when Ship2, Filamin or Lamellipodin were individually knocked down. We also observed decreased lamellipodia formation and cell migration in these knock down cells. These data provide evidence that Nephrin not only initiates actin polymerization but also assembles a protein complex that is necessary to regulate the architecture of the generated actin filament network and focal adhesion dynamics.

Venkatareddy, Madhusudan; Cook, Leslie; Abuarquob, Kamal; Verma, Rakesh; Garg, Puneet



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


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

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



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.



Human cathepsin B1. Inhibition by ?2-macroglobulin and other serum proteins  

PubMed Central

1. Normal human serum was found to inhibit human cathepsin B1. 2. The major inhibitor present in serum was purified and identified as ?2-macroglobulin. 3. ?2-Macroglobulin was found to bind cathepsin B1 in an approximately 1:1 molar ratio. When bound, the enzyme retained about 50% of its proteolytic activity, and up to 80% of its activity against ?-N-benzoyl-dl-arginine 2-naphthylamide. 4. Pretreatment of ?2-macroglobulin with cathepsin B1 inactivated by exposure to pH8.5 or iodoacetic acid, in large molar excess, did not prevent the subsequent binding of active enzyme. Active enzyme, once bound, was not protected from inhibition by 1-chloro-4-phenyl-3-tosylamido-l-butan-2-one. 5. Cathepsin B1 was also inhibited by human immunoglobulin G, at high concentration. 6. Because it had been suggested that haptoglobin is responsible for the inhibition of `cathepsin B' by serum, a method was devised for the selective removal of haptoglobin from mixtures of serum proteins by adsorption on haemoglobin covalently linked to Sepharose. No evidence was obtained that haptoglobin has any inhibitory activity against the enzyme. ImagesPLATE 1

Starkey, Phyllis M.; Barrett, Alan J.



Serum phospholipid transfer protein activity after a high fat meal in patients with insulin-treated type 2 diabetes.  


Plasma phospholipid transfer protein (PLTP) mediates both net transfer and exchange of phospholipids between different lipoproteins. Animal studies have shown that it is closely related to the development of atherosclerosis. Although many studies have indicated that PLTP activity is increased in diabetes mellitus, the role of PLTP in diabetes is still unclear. To evaluate the influence of a high-fat meal on PLTP activity, 50 nondiabetic patients with coronary heart disease (CHD), 50 insulin-treated Type 2 diabetics, and 50 healthy controls were included. We determined PLTP activity before and 4 and 8 h after a high-fat meal. As expected, serum PLTP activity was significantly higher in CHD patients than in healthy controls (71.0 +/- 46.2 vs. 54.0 +/- 33.8 pmol/microl/h, P = 0.032) at baseline. More importantly, we found that serum PLTP activity increased to its maximum 4 h after fat loading and then decreased to nearly basal levels after 8 h both in controls and CHD patients. In contrast, PLTP activity continuously increased during this time period in the diabetic patients. With regards to the data from this study we hypothesize that serum PLTP is involved in the clearance of postprandial lipoproteins and this process is attenuated in diabetes. Since postprandial lipoproteins are atherogenic, the delay in clearance of these particles could play an important role in the development of atherosclerosis in patients with diabetes mellitus. PMID:20108050

Schlitt, Axel; Schwaab, Bernhard; Fingscheidt, Kirsten; Lackner, Karl J; Heine, Gunnar H; Vogt, Alexander; Buerke, Michael; Maegdefessel, Lars; Raaz, Uwe; Werdan, Karl; Jiang, Xian-Cheng




EPA Science Inventory

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


Isolation of serum protein organometallic corrosion products from 316LSS and HS-21 in vitro and in vivo.  


An investigation into blood-borne organometallic compounds that arise from the corrosion of metals used in orthopedic prosthetic devices was conducted using an in vivo rat model with an implantation time of 10 days and an in vitro human serum model with an incubation time of 5 days. Both models involved 316LSS and HS-21 in the spherical powder form of 55 +/- 5 microns microns in diameter at three different surface areas to body weight ratios. Gel chromatography on cross-linked dextran (G-200) was used to fractionate the serum proteins which bound the metal ions (chromium, cobalt, and nickel) released and identify them. Atomic absorption-spectrophotometry analysis measured the concentration of the metal ions in each serum protein peak as well as whole serum from both models, and red cell and tissue from the in vivo model. Within the serum proteins, the metal ions were bound to two of the principal serum protein peaks. Similar distributions of the metals among the serum protein peaks were not noted. PMID:6699034

Woodman, J L; Black, J; Jiminez, S A



Serial Measurements of Serum Protein, Glycoprotein, and Lipoprotein Fractions in Normal and Venezuelan Equine Encephalomyelitis-Vaccinated Ponies and Burros.  

National Technical Information Service (NTIS)

Simultaneous comparisons of serum protein, glycoprotein, and lipoprotein on cleared cellulose acetate are described in normal ponies and burros before and after vaccination with Venezuelan equine encephalomyelitis (vee) attenuated virus. Slight quantitati...

J. B. Rollins R. H. Fiser T. D. Shultz



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.

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



Antibody array-based technologies for cancer protein profiling and functional proteomic analyses using serum and tissue specimens  

Microsoft Academic Search

In the context of other proteomic technologies, targeted antibody arrays are strongly contributing for protein profiling and\\u000a functional proteomics analyses in serum specimens. Protein-protein interactions, post-translational modifications, and interaction\\u000a between protein and DNA or RNA can all shift the activity of a protein from what would have been predicted by its level of\\u000a transcription. Functional proteomics studies the interaction of

Marta Sanchez-Carbayo



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


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

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



Release of Glial Tissue-Specific Proteins After Acute Stroke A Comparative Analysis of Serum Concentrations of Protein S-100B and Glial Fibrillary Acidic Protein  

Microsoft Academic Search

Background and Purpose—This study was aimed at the comparative analysis of serum concentrations of glial fibrillary acidic protein (GFAP) and protein S-100B in patients with acute stroke. Methods—We investigated 32 patients with stroke symptoms consistent with cerebral ischemia in the anterior territory of vascular supply. Serial venous blood samples were taken after admission to the hospital and during the first

Manfred Herrmann; Pieter Vos; Michael T. Wunderlich; Chris H. M. M. de Bruijn; Karl J. B. Lamers


Effects of dextran on five biuret-based procedures for total protein in serum.  


We evaluated the effect of dextran on values for total protein in serum as measured by the biuret method with five widely used automated instruments: the American Monitor Parallel; the Du Pont aca II; the Roche Cobas-Bio; the Kodak Ektachem 400; and the Beckman Astra 8. Dextran concentrations as great as 25 or 30 g/L had relatively little or no influence on total protein measurements by the latter three instruments. Dextran concentrations exceeding 6 g/L caused falsely low results with the aca, whereas the Parallel gave falsely high results when the dextran concentration exceeded 2 g/L. The aca total protein procedure could be protected from the interference by dextran concentrations up to 30 g/L by injecting 0.4-0.8 mL of ethylene glycol directly into the reagent pack before sampling. However, we could not eliminate the interference with the Parallel procedure by any simple means; we thus recommend that it not be used for measuring total protein in serum samples from patients who are being treated with dextran. PMID:2415271

Barnes, D B; Pierce, G F; Lichti, D; Landt, M; Koenig, J; Chan, K M



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

PubMed Central

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

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



[Research on detecting concentration of serum protein based on resonance Rayleigh scattering].  


The resonance Rayleigh scattering spectral detection system was designed based on the 2, 9, 16, 23-tetracarboxylate-phthalocyanine zinc and protein system. In the system, excitation light source is 405 nm wide band gap semiconductor lasers, and monochromator is 475 nm narrow-band band-pass filter, and the detector is low-noise and high-gain photoelectric amplifier based on blue-ray enhanced photodiode. Experiment shows that, the solution's strong absorption wavelength is near 420 nm. Under the action of incentive light, resonance Rayleigh scattering is generated at the resonant wavelength, and the scattering intensity is proportional to the protein content. The system uses 2, 9, 16, 23-tetracarboxylate as the spectrum probe to determine the concentration of serum proteins by resonance Rayleigh scattering method. Its linear detection range is 10 - 50 mg.mL-1, and its detection limit is 0. 001 mg.mL-1. The newly developed device for detecting concentration of the serum protein has the advantages of small size, low cost, low power consumption, and being easy to use. PMID:23705447

Wang, Gao; Feng, Qiao-Ling; Xue, Zhong-Jin; Li, Yang-Jun; Zhou, Han-Chang



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

Microsoft Academic Search

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

Kyle W. Selcer; Brent D. Palmer



Profiling of serum and tissue high abundance acute-phase proteins of patients with epithelial and germ line ovarian carcinoma  

Microsoft Academic Search

BACKGROUND: Acute-phase response involves the simultaneous altered expression of serum proteins in association to inflammation, infection, injury or malignancy. Studies of the acute-phase response usually involve determination of the levels of individual acute-phase serum proteins. In the present study, the acute-phase response of patients with epithelial (EOCa) and germ-line (GOCa) ovarian carcinoma was investigated using the gel-based proteomic approach, a

Yeng Chen; Boon-Kiong Lim; Suat-Cheng Peh; Puteri Shafinaz Abdul-Rahman; Onn Haji Hashim



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

Microsoft Academic Search

Vitamin D-binding protein (DBP) and albumin (ALB) are abundant serum proteins and both possess high-affinity binding for saturated and unsaturated fatty acids. However, certain differences exist. We surmised that in cases where serum albumin level is low, DBP presumably can act as a transporter of fatty acids. To explore this possibility we synthesized several alkylating derivatives of 14C-palmitic acid to

Narasimha Swamy; Rahul Ray



Nonspecific Precipitation of Serum Proteins by Sodium Lauryl Sulfate in Agar Diffusion and Immunoelectrophoresis  

PubMed Central

The anionic detergent sodium lauryl sulfate (SLS), in a final concentration of 0.1% and greater, reacted with whole serum in agar diffusion and immunoelectrophoresis to form artifactual precipitin lines. These lines occurred when either Ionagar or agarose was used as the supporting gel and were not affected by the presence of urea and 2-mercaptoethanol. Analytic chemical tests confirmed that the precipitating agent is SLS, and staining techniques showed that the detergent precipitates both protein and lipoprotein components of whole serum. Multiple artifactual precipitin lines occurred with a wide variety of animal sera, and a single line formed with human 7S immunoglobulin. Hence, in agar diffusion studies in which SLS is present in the test system, these artifactual lines may be easily misinterpreted as true antigen-antibody precipitin reactions. Images

Palmer, E. L.; Martin, M. L.; Hierholzer, J. C.; Ziegler, D. W.



Isolation and characterization of C-reactive protein and serum amyloid P component in the rat.  

PubMed Central

C-reactive protein (RP) and serum amyloid P component (SAP) have been identified for the first time in rat serum and isolated by calcium-dependent affinity chromatography. Rat CRP closely resembled human CRP in its amino acid composition, in having five subunits per molecule and in its electron microscopic appearance as a pentameric annular disc. It differed, however, from all other mammalian CRP's characterised hitherto in being a glycoprotein bearing a single complex oligosaccharide on each polypeptide subunit. Furthermore one pair of tis subunits per molecule was linked by a interchain disulphide bridges whereas in other animals the subunits of both CRP and SAP are all non-covalently associated. The serum concentration of CRP in normal healthy laboratory rats and in specific pathogen-free rats was 300-600 micrograms/ml which is much greater than has been described in any other species and exceeds even maximal acute phase levels of CRP in man. Following injections of casein or croton oil, serum CRP levels rose to a maximum of about 900 micrograms/ml. Rat CRP bound to pneumococcal C-polysaccharide (CPS( but, in marked contrast to the behaviour of CRP from man, rabbit and marine teleost fish, it did not precipitate with CPS solutions, agglutinate CPS-coated sheep erythrocytes or initiate complement activation. Rat SAP, like SAP of other species, was a glycoprotein but unlike them it was composed only of a single pentameric disc not two such discs interacting face-to-face. The normal level of SAP in rat serum was 20-50 micrograms/ml, very similar to the levels seen in man, and it did not behave as an acute phase reactant in response to casein or croton-oil injections. In this respect it resembled human SAP but differed from murine SAP which is a major acute phase reactant. Images Figure 2 Figure 3 Figure 5 Figure 6 Figure 7 Figure 8 Figure 11

de Beer, F C; Baltz, M L; Munn, E A; Feinstein, A; Taylor, J; Bruton, C; Clamp, J R; Pepys, M B



The OspE-related proteins inhibit complement deposition and enhance serum resistance of Borrelia burgdorferi, the lyme disease spirochete.  


Borrelia burgdorferi, the Lyme disease spirochete, binds the host complement inhibitors factor H (FH) and FH-like protein 1 (FHL-1). Binding of FH/FHL-1 by the B. burgdorferi proteins CspA and the OspE-related proteins is thought to enhance resistance to serum-mediated killing. While previous reports have shown that CspA confers serum resistance in B. burgdorferi, it is unclear whether the OspE-related proteins are relevant in B. burgdorferi serum resistance when OspE is expressed on the borrelial surface. To assess the role of the OspE-related proteins, we overexpressed them in a serum-sensitive CspA mutant strain. OspE overexpression enhanced serum resistance of the CspA-deficient organisms. Furthermore, FH was more efficiently bound to the B. burgdorferi surface when OspE was overexpressed. Deposition of complement components C3 and C5b-9 (the membrane attack complex), however, was reduced on the surface of the OspE-overexpressing strain compared to that on the CspA mutant strain. These data demonstrate that OspE proteins expressed on the surface of B. burgdorferi bind FH and protect the organism from complement deposition and subsequent serum-mediated destruction. PMID:21282413

Kenedy, Melisha R; Akins, Darrin R



Proteomic expression profiling and identification of serum proteins using immobilized trypsin beads with MALDI-TOF/TOF.  


MALDI-TOF mass spectrometry is a widely used technique for serum protein expression profiling and biomarker discovery. Many profiling strategies typically employ chemical affinity beads or surfaces to decrease sample complexity of dynamic fluids such as serum or plasma. However, many of the proteins captured on a particular surface or bead are not resolved in the lower mass ranges where time-of-flight mass spectrometers are most effective. Thus, a majority of reported protein expression profiling studies primarily interrogate the native low molecular mass constituents of the target sample. We report an expression profiling workflow that utilizes immobilized trypsin paramagnetic beads following an initial affinity bead fractionation step, thereby reducing large mass proteins to peptides that are better suited to analysis and sequencing determinations. Our bead-based trypsin approach resulted in more efficient digestion of complex serum protein extracts at short incubation times. This method was reproducible and readily adaptable to robotic sample handling and may be combined in tandem with other bead fractionation surfaces. When weak cationic and weak anionic bead surfaces were used, experimental conditions were optimized for tandem combinations of these beads with the immobilized trypsin step to produce an efficient serum fractionation strategy. A proof-of-concept pilot experiment using pooled human serum samples demonstrating reproducibility is presented, along with the sequence determination of selected tryptic peptides of serum proteins. PMID:19603828

Karbassi, Izabela D; Nyalwidhe, Julius O; Wilkins, Christopher E; Cazares, Lisa H; Lance, Raymond S; Semmes, O John; Drake, Richard R



Serum insulin-like growth factor-I, iron, C-reactive protein, and serum amyloid A for prediction of outcome in dogs with pyometra.  


Pyometra, accumulation of pus in the uterus, is a bacterial infection that frequently initiates systemic inflammation. The disease may have lethal consequences when the systemic effects are severe or complications occur. Markers for identifying high-risk patients and predicting outcome are therefore in high demand. The objective of this study was to measure serum concentrations of insulin-like growth factor-I (IGF-I), iron, C-reactive protein (CRP), and serum amyloid A (SAA) in bitches with pyometra and to explore the possible value of these variables for detection of increased morbidity. In total, 31 bitches were diagnosed with pyometra and destined for surgical treatment (ovariohysterectomy) and 17 healthy bitches were included in the study. Concentrations of IGF-I and iron were lower in the pyometra group (mean concentration 221.2 ± 22.5 ng/mL and 16.9 ± 1.6 ?mol/L, respectively) compared with the healthy control group (mean concentration 366.7 ± 46.2 ng/mL and 38.1 ± 2.7 ?mol/L, respectively). In contrast, concentrations of CRP and SAA were significantly higher in bitches with pyometra (mean concentrations 212.9 ± 17.3 mg/L and 119.9 ± 8.5 mg/L, respectively) compared with the control group (<5 mg/L and <10 mg/L, respectively). None of the explored variables were associated with morbidity as measured by duration of postoperative hospitalization. In conclusion, IGF-I and iron concentrations were decreased in pyometra, whereas SAA and CRP concentrations were increased in the disease. Although unspecific, measurement of these variables may be valuable as adjunctive markers for prognosis in cases of pyometra. PMID:24661434

Jitpean, Supranee; Holst, Bodil Ström; Höglund, Odd V; Pettersson, Ann; Olsson, Ulf; Strage, Emma; Södersten, Fredrik; Hagman, Ragnvi



Interaction of silver nanoparticles with serum proteins affects their antimicrobial activity in vivo.  


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; Chakravortty, Dipshikha



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.

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



A new sandwich immunoassay for detection of the ?-secretase cleaved, soluble amyloid-? protein precursor in cerebrospinal fluid and serum.  


Alzheimer's disease (AD) is the most common neurodegenerative disorder. Frequently used diagnostic biomarkers are amyloid-?42 (A?42), tau, and phospho-tau, which are measured in cerebrospinal fluid (CSF), and allow a reasonable, but not full, separation of AD patients and controls. Besides A?42, additional proteolytic cleavage products of the amyloid-? protein precursor (A?PP) have been investigated as potential biomarkers. This includes the ?-secretase cleaved soluble A?PP ectodomain (sA?PP?). However, some studies found a reduction of sA?PP?, whereas other studies reported an increase of sA?PP? in the CSF of AD patients. The divergent findings may result from the detection of sA?PP? with antibodies, such as 6E10, which do not exclusively detect sA?PP?, but also the alternative ?-secretase cleavage product sA?PP?'. Here, we used the sA?PP?-specific antibody 14D6 and developed an ELISA-like sandwich immunoassay. The assay specifically detected sA?PP? in cell culture supernatants, in human CSF and even in serum, which is more readily accessible than CSF. The assay was used to analyze sA?PP? levels in CSF and serum of AD patients and controls. The assay detected a mild, but significant increase in sA?PP? in the CSF of AD patients compared to non-demented controls, while a mild reduction was observed in serum. The 14D6 assay in CSF allowed a better separation of AD patients from controls compared to the 6E10 antibody. Taken together, the new assay is widely applicable for specific sA?PP? measurement in culture media, CSF, and serum. PMID:23948916

Taverna, Mara; Straub, Tobias; Hampel, Harald; Rujescu, Dan; Lichtenthaler, Stefan F



Serum protein, ascorbic acid & iron & tissue collagen in oral submucous fibrosis--a preliminary study.  


A study of 36 patients with oral submucous fibrosis, revealed that all patients had the habit of chewing betel nut, pan masala or the traditional mixture (betel nut, betel leaf and lime) suggesting a link between fibrosis and arecanut. There was an increase in the globulin fraction of protein and hence a decreased A/G ratio in these patients. There was a significant increase in total protein levels possibly due to the increase in globulin fractions and other serum proteins. Ascorbate and iron levels decreased perhaps because of their utilisation in collagen synthesis. The total tissue collagen content increased significantly in patients with advanced disease and, it increased with the progression of the disease leading to hypomobility of the tongue, lips, cheeks, soft palate and faucial pillars. PMID:8225452

Anuradha, C D; Devi, C S



Impact of hyperthermic intraperitoneal chemotherapy on Hsp27 protein expression in serum of patients with peritoneal carcinomatosis.  


Despite the strong rationale for combining cytoreductive surgery (CRS) with hyperthermic intraperitoneal chemotherapy (HIPEC) in patients with peritoneal carcinomatosis, thermotolerance and chemoresistance might result from heat shock protein overexpression. The aim of the present study was thus to determine whether the heat shock protein 27 (Hsp27), a potential factor in resistance to treatment, could have a higher level in serum from patients under this combined therapy. Patients receiving CRS plus HIPEC for peritoneal carcinomatosis (group 1), patients with cancer or a history of cancer undergoing abdominal surgery (group 2), and patients without malignancies undergoing abdominal surgery (group 3) were included. Hsp27 serum levels were determined before and at different times following CRS and HIPEC using enzyme-linked immunosorbent assay. In group 1 (n = 25), the high Hsp27 levels, observed at the end of surgery compared with before (p < 0.0001), decreased during HIPEC, but remained significantly higher than before surgery (p < 0.0005). In groups 2 (n = 11) and 3 (n = 15), surgery did not significantly increase Hsp27 levels. A targeted molecular strategy, inhibiting Hsp27 expression in tumor tissue, could significantly reduce resistance to the combined CRS plus HIPEC treatment. This approach should be further assessed in a clinical phase I trial. PMID:23508575

Kepenekian, Vahan; Aloy, Marie-Thérčse; Magné, Nicolas; Passot, Guillaume; Armandy, Emma; Decullier, Evelyne; Sayag-Beaujard, Annie; Gilly, François-Noël; Glehen, Olivier; Rodriguez-Lafrasse, Claire



Association of Genetic Variants of Casein and Milk Serum Proteins with Milk, Fat, and Protein Production by Dairy Cattle  

Microsoft Academic Search

Polyacrylamide gel electrophoresis methods were used to phenotype caseins for 2045 cows and milk serum proteins for 3870 cows distributed in 63 Quebec dairy herds. Frequencies were: %l-casein A.003, asl-casein B.970, C~sl-easein C .027; J3-casein A1 .561, ~3-casein A2 .421, J3-casein A3 .011, 13-casein B.007 ; K-casein A.744, K-casein B.256 ; ~-lactoglobulin A .387, \\/3-1actoglobulin B.613; a-lactal- burnin B 1.00.

K. F. Ng-Kwai-Hang; J. F. Hayes; J. E. Moxley; H. G. Monardes



Feed consumption, nutrient utilization and serum metabolite profile of captive blackbucks (Antelope cervicapra) fed diets varying in crude protein content.  


A feeding trial was conducted to determine the optimum level of crude protein (CP) in the diet of captive blackbuck (Antelope cervicapra) in which feed consumption and nutrient utilization are maximal. Fifteen blackbucks (BW 25-34 kg) were distributed into three groups of five each in an experiment of 75-days duration including a digestion trial of 5-day collection period. All the animals were offered 200 g of concentrates and fresh maize fodder ad libitum. The overall CP content of the three respective diets was 6.9%, 10.4% and 12.7%. Blood samples were collected on the last day of the experiment. Intake and digestibility of CP increased (p < 0.01) with the increased level of CP in the diet. Feed consumption and nutrient intake were not significantly different among the groups. However, digestibilities of most of the nutrients were higher in the 10.4% CP diet than in the 6.9% CP diet. The endogenous loss of nitrogen was similar among the groups. Based on the endogenous losses, minimum N requirement was calculated to be 776 mg/kg BW(0.75) /day, and to meet this requirement, diet must contain at least 8.27% CP. Serum urea nitrogen concentration increased (p < 0.01) with increased level of dietary CP. Serum level of glutamate oxaloacetate transaminase and glutamate pyruvate transaminase was higher (p < 0.05) in the group fed 6.9% CP diet. Animals in the group fed low protein diet also lost body mass during the experimental period. It was concluded that a diet containing 10.4% CP was optimum for maximizing nutrient utilization without any adverse effect on voluntary feed consumption and serum metabolite profile of blackbucks. PMID:21585563

Das, A; Katole, S; Kumar, A; Gupta, S P; Saini, M; Swarup, D



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.

Rysz, Jacek; Paradowski, Marek



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

PubMed Central

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

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



Postmortem serum protein S100B levels with regard to the cause of death involving brain damage in medicolegal autopsy cases.  


The protein S100 is an acidic calcium-binding protein, and the subunit S100B is the most abundantly found in the brain. The aim of the present study was a comprehensive analysis of serum S100B levels in medicolegal autopsy cases (within 48 h postmortem, n = 283) including victims with head and non-head injuries and also non-injury fatalities with regard to the cause of death involving brain damage. The serum level was usually higher in the subclavian vein than in the right heart and external iliac vein, and the lowest in the left heart blood, showing no significant postmortem influence. The serum level in the right heart and subclavian vein was markedly higher for acute deaths from head injury and asphyxiation due to neck compression (strangulation and hanging), and moderately and mildly elevated for other blunt and sharp instrument injury cases, respectively. For head injury, the serum levels were lower for subacute deaths than for acute deaths. These observations suggest that the elevation of serum S100B may mainly be caused by leakage following massive brain damage due to injury and cerebral hypoxia/ischemia, and in part by systemic hypoxic/traumatic tissue damage, depending on the survival time. PMID:16337822

Li, Dong-Ri; Zhu, Bao-Li; Ishikawa, Takaki; Zhao, Dong; Michiue, Tomomi; Maeda, Hitoshi



Evaluation of time-resolved fluoroimmunoassay with Eu-labelled protein-A for serum progesterone.  


Progesterone has been assayed in several serum samples by a time-resolved fluoroimmunoassay (TR-FIA). The solid phase was 11 alpha-hydroxyprogesterone hemisuccinate bound covalently to ovalbumin and adsorbed on wells of polystyrene. The assay was based on competitive reaction of solid phase-bound hormone and samples with specific antibody labelled in situ with protein-A prelabelled with europium. The bound Eu was dissociated from the solid phase by an enhancement solution and measured by time-resolved fluorometry. The coefficient of correlation between TR-FIA and RIA was 0.97. PMID:2668643

Ius, A; Ferrara, L; Meroni, G; Bacigalupo, M A



Serum S-100B protein monitoring in patients with severe traumatic brain injury  

Microsoft Academic Search

Objective  S-100B protein is a promising marker of injury severity and outcome after head injury. We examined the relationship between\\u000a serum S-100B concentrations and injury severity, clinical course, survival, and treatment efficacy after severe traumatic\\u000a brain injury (TBI).\\u000a \\u000a \\u000a \\u000a Design and setting  Prospective observational study in a neurosurgical intensive care unit.\\u000a \\u000a \\u000a \\u000a Patients and participants  102 adult patients with severe TBI, admitted between June 2001 and November

Stefanos Korfias; George Stranjalis; Efstathios Boviatsis; Christina Psachoulia; Gerard Jullien; Barbara Gregson; A. David Mendelow; Damianos E. Sakas



The effect of serum protein concentration on wear rates in a hip simulator.  


Ultra high molecular weight polyethylene cups, 22 mm in diameter, were aged for 5 years in the normal laboratory environment. Half of the samples had been processed by the standard radiation sterilization techniques, while the remainder had been cross-linked by a technique involving higher radiation doses and controlled temperature at the time of irradiation. The samples had been tested in a hip simulator for 5,000,000 cycles using a lubricant that had been diluted 1:1 with deionized water. Once that testing was completed, further testing was conducted using lubricant with greater and lesser serum protein concentrations, and the results compared with those that had already been recorded. Comparison of the wear rates within the study as well as to published data concerning the effect of serum concentration showed results that were consistent with assumed differences in lubrication ability at different concentrations. The results of other published studies were found to be inconsistent with each other and different from some of the results of this study. There is shown to be a need for carefully controlled and conducted studies to agree, if possible, on the importance of the serum concentration and the appropriate parameters to be used in testing, as well as variations that may be necessary with different bearing material characteristics. PMID:19833674

St John, Kenneth



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

PubMed Central

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

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



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



Effect of soy protein varying in isoflavone content on serum lipids in healthy young men1-3  

Microsoft Academic Search

Background: Previous research supports a role for soy protein in reducing serum lipids; however, few studies involved healthy male subjects or focused on soy isoflavones (or did both). Objective: The objective was to ascertain the effects of soy protein varyinginisoflavonecontentonserumlipidsinhealthyyoungmen. Design: Thirty-five males (x SD age: 27.9 5.7 y) consumed milk protein isolate (MPI), low-isoflavone soy protein isolate (low- iso

Brianne L McVeigh; Barbara L Dillingham; Johanna W Lampe; Alison M Duncan


New protein structure model evaluation methods that include a side-chain consensus score for the protein modeling.  


Selecting the best quality model from a set of predicted structures is one of the most important aspects of protein structure prediction. We have developed model quality assessment programs that select high quality models which account for both the Calpha backbone and side-chain atom positions. The new methods are based on the consensus method with consideration of the side-chain environment of a protein structure and the secondary structure agreement. This Side-chain Environment Consensus (SEC) method is compared with the conventional consensus method, 3D-Jury (Ginalski K. et al., Bioinformatics, 19, 1015-1018 (2003)), which takes into account only the Calpha backbone atoms of the protein model. As the result, it was found that the SEC method selects the models with more accurate positioning of the side-chain atoms than the 3D-Jury method. When the SEC method was used in combination with the 3D-Jury method (3DJ+SEC), models were selected with improved quality both in the Calpha backbone and side-chain atom positions. Moreover, the CIRCLE (CCL) method (Terashi G. et al., Proteins, 69 (Suppl. 8), 98-107 (2007)) based on the 3D-1D profile score has been shown to select the best possible models that are the closest to the native structures from candidate models. Accordingly, the 3DJ+SEC+CCL method, in which CIRCLE is used after reducing the number of candidates by the 3DJ+SEC consensus method, was found to be very effective in selecting high quality models. Thus, the best method (the 3DJ+SEC+CCL method) includes the consensus approaches of the Calpha backbone and the side-chains, the secondary structure agreement and the 3D-1D profile score which corresponds to the free energy-like score in the residues of the protein model. In short, new algorithms are introduced in protein structure evaluation methods that are based on a side-chain consensus score. Additionally, in order to apply the 3DJ+SEC+CCL method and indicate the usefulness of this method, a model of human Cabin1, a protein associated with p53 function and cancer, is created using various internet modeling and alignment servers. PMID:20118576

Kanou, Kazuhiko; Hirata, Tomoko; Terashi, Genki; Umeyama, Hideaki; Takeda-Shitaka, Mayuko



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

PubMed Central

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

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



Association between serum levels of C-reactive protein and personality traits in women  

PubMed Central

Background While low-grade inflammation has consistently been observed in subjects with depression, studies on the possible relationship between inflammation and other aspects of brain function are as yet sparse. In this study, we aimed to investigate the possible association between serum levels of the inflammation marker C-reactive protein (CRP) and personality traits. Methods In this study, serum levels of high-sensitivity CRP were determined by ELISA in a population of 270 42-year-old women recruited from the population registry who had been assessed using the Temperament and Character Inventory. Self-reported previous or ongoing depression was also recorded. Unpaired two-tailed t-tests were used for comparison between two groups and correlations were evaluated by the calculation of Pearson's r-coefficient. Results The temperament trait harm avoidance was positively (r = 0.227, p < 0.05) and the character trait self-directedness was negatively (r = -0.261, p < 0.01) associated with serum levels of CRP (p-values corrected for multiple comparisons). The correlations between the personality traits and CRP were observed also after exclusion of subjects reporting ongoing depression (n = 26). Whereas women reporting ongoing depression showed significantly increased levels of CRP as compared to non-depressed women (n = 155), women reporting a history of depression displayed no significant difference in CRP levels as compared to women that reported that they had never been depressed. Conclusion Serum levels of CRP in women was found to be associated with the personality traits harm avoidance and self-directedness. In addition, moderately elevated levels may be a state dependent marker of depression.

Henningsson, Susanne; Baghaei, Fariba; Rosmond, Roland; Holm, Goran; Landen, Mikael; Anckarsater, Henrik; Ekman, Agneta



Serum C-reactive protein and procalcitonin levels in non-small cell lung cancer patients  

PubMed Central

Aim of the study The basic uses of C-reactive protein (CRP) and procalcitonin (PCT) in clinical practice are in the diagnosis and follow-up of infectious disease. The fact that CRP already achieves high levels in cases with lung cancer, however, limits its diagnostic specificity. Procalcitonin may be an important marker in the differential diagnosis of lung cancer patients who have fever and high CRP levels. Our objective in this study was to determine the levels of CRP and PCT in patients with newly diagnosed non-infectious non-small cell lung cancer (NSCLC) and to relate these results to patient and disease characteristics. Material and methods Serum CRP and PCT levels were measured in 79 histopathologically proven NSCLC patients and 20 healthy controls. Results were compared with demographic and clinical variables in patients with NSCLC. Results Serum CRP concentrations were significantly higher in NSCLC patients compared to the control group [38.30 (7.79–185) mg/dl vs. 7.79 (3.36–26.10) mg/dl; p < 0.001]. There was no significant difference between the two groups in PCT levels (p > 0.05). A mild, positive correlation was found between CRP level and tumor diameter. When comparing CRP levels in the lung cancer patients grouped according to age, sex, smoking status, clinical TNM staging and performance status (PS), the only significant difference found was that for PS score. Conclusions High serum CRP levels in non-infectious NSCLC patients are mainly related to PS status and weakly to tumor size. Adding serum PCT measurement may contribute to exclusion of infections in patients with NSCLC.

Koylu, Habibe; Kanat, Fikret; Arslan, Ugur; Ozer, Faruk



A proteomic study of serum from children with autism showing differential expression of apolipoproteins and complement proteins.  


Modern methods that use systematic, quantitative and unbiased approaches are making it possible to discover proteins altered by a disease. To identify proteins that might be differentially expressed in autism, serum proteins from blood were subjected to trypsin digestion followed by liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) on time-of-flight (TOF) instruments to identify differentially expressed peptides. Children with autism 4-6 years of age (n=69) were compared to typically developing children (n=35) with similar age and gender distributions. A total of 6348 peptide components were quantified. Of these, five peptide components corresponding to four known proteins had an effect size >0.99 with a P<0.05 and a Mascot identification score of 30 or greater for autism compared to controls. The four proteins were: Apolipoprotein (apo) B-100, Complement Factor H Related Protein (FHR1), Complement C1q and Fibronectin 1 (FN1). In addition, apo B-100 and apo A-IV were higher in children with high compared to low functioning autism. Apos are involved in the transport of lipids, cholesterol and vitamin E. The complement system is involved in the lysis and removal of infectious organisms in blood, and may be involved in cellular apoptosis in brain. Despite limitations of the study, including the low fold changes and variable detection rates for the peptide components, the data support possible differences of circulating proteins in autism, and should help stimulate the continued search for causes and treatments of autism by examining peripheral blood. PMID:17189958

Corbett, B A; Kantor, A B; Schulman, H; Walker, W L; Lit, L; Ashwood, P; Rocke, D M; Sharp, F R



Serum albumin prevents protein aggregation and amyloid formation and retains chaperone-like activity in the presence of physiological ligands.  


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

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



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

PubMed Central

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

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



Human serum albumin binding to silica nanoparticles - effect of protein fatty acid ligand.  


Neutron reflectivity shows that fatted (F-HSA) and defatted (DF-HSA) versions of human serum albumin behave differently in their interaction with silica nanoparticles premixed in buffer solutions although these proteins have close to the same surface excess when the silica is absent. In both cases a silica containing film is quickly established at the air-water interface. This film is stable for F-HSA at all relative protein-silica concentrations measured. This behaviour has been verified for two small silica nanoparticle radii (42 Ĺ and 48 Ĺ). Contrast variation and co-refinement have been used to find the film composition for the F-HSA-silica system. The film structure changes with protein concentration only for the DF-HSA-silica system. The different behaviour of the two proteins is interpreted as a combination of three factors: increased structural stability of F-HSA induced by the fatty acid ligand, differences in the electrostatic interactions, and the higher propensity of defatted albumin to self-aggregate. The interfacial structures of the proteins alone in buffer are also reported and discussed. PMID:24595605

Ang, Joo Chuan; Henderson, Mark J; Campbell, Richard A; Lin, Jhih-Min; Yaron, Peter N; Nelson, Andrew; Faunce, Thomas; White, John W



Serum Oxidized Protein and Prostate Cancer Risk within the Prostate Cancer Prevention Trial  

PubMed Central

To evaluate the role of oxidative stress in prostate cancer risk, we analyzed serum levels of protein carbonyl groups in 1808 prostate cancer cases and 1805 controls, nested in the Prostate Cancer Prevention Trial, a randomized, placebo-control trial that found finasteride decreased prostate cancer risk. There were no significant differences in protein carbonyl levels in baseline samples between those later diagnosed with prostate cancer and those without at the end of study biopsy. Adjusted ORs and 95% CIs for the 4th quartile of protein carbonyl level for the combined, placebo and finasteride arms were 1.03 (95% CI 0.85–1.24), 0.88 (95% CI 0.69–1.12) and 1.27 (95% CI 0.94–1.71), respectively. There were no significant associations between carbonyl level and risk when analyzing high- and low-grade disease separately, nor did finasteride impact protein oxidation levels. The results of this large nested case-control study do not support the hypothesis that oxidative stress, at least as measured by protein carbonyl level, plays a role in prostate cancer.

Hoque, Ashraful; Ambrosone, Christine B.; Till, Cathee; Goodman, Phyllis J.; Tangen, Cathy; Kristal, Alan; Lucia, Scott; Wang, Qiao; Kappil, Maya; Thompson, Ian; Hsing, Ann W.; Parnes, Howard; Lippman, Scott M.; Santella, Regina M.



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


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

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



Interplay between promoter and structural gene variants control basal serum level of mannan-binding protein.  


Mannan-binding protein (MBP) is a serum lectin participating in the innate immune defense by opsonizing various microorganisms for phagocytosis. Opsonization defect due to MBP deficiency and low levels of the protein can partially be explained by the dominant effect of three different mutations in the structural part of the MBP gene. Large interracial differences in the frequencies of these variants have previously been described, but they cannot explain the large interindividual variation in MBP serum concentration. We describe the existence of additional polymorphisms at positions -550 (H/L variants) and -221 (X/Y variants) in the promoter region of the gene. The promoter haplotypes, HY, LY, and LX, show associations with high, medium, and low levels of MBP serum concentrations, respectively. Moreover, this represents a genetic system with additive effect of haplotypes in which a low producing LX haplotype in the homozygous state down-regulates the basal expression of MBP as effectively as a single structural variant. Populations of pure Eskimos, Caucasoids, and black Africans show marked interethnic differences in the frequencies of promoter haplotypes regulating the expression of the normal peptide, with the HY haplotype frequency varying from 0.83 in Eskimos via 0.33 in Caucasoids to 0.08 in Africans. The LY haplotype frequency varies from 0.04 in Eskimos via 0.39 in Caucasoids to 0.23 in Africans. The LX haplotype frequency varies from 0.03 in Eskimos via 0.24 in Caucasoids to 0.23 in Africans. The effect of the promoter variants can explain almost all of the ethnic differences not explainable by the structural variants alone. PMID:7673719

Madsen, H O; Garred, P; Thiel, S; Kurtzhals, J A; Lamm, L U; Ryder, L P; Svejgaard, A



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

Microsoft Academic Search

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

Heping Zhou; Kerri Bouwman; Mark Schotanus; Cornelius Verweij; Jorge A Marrero; Deborah Dillon; Jose Costa; Paul Lizardi; Brian B Haab



A Serum Protein-Based Algorithm for the Detection of Alzheimer's Disease  

PubMed Central

Background Alzheimer's disease (AD) is the most common form of age-related dementia and one of the most serious health problems in the industrialized world. Biomarker approaches to diagnostics would be more time and cost effective and may also be useful for identifying endophenotypes within AD patient populations. Methods We analyzed serum protein-based multiplex biomarker data from 197 patients diagnosed with AD and 203 controls from a longitudinal study of Alzheimer's disease being conducted by the Texas Alzheimer's Research Consortium to develop an algorithm that separates AD from controls. The total sample was randomized equally into training and test sets and random forest methods were applied to the training set to create a biomarker risk score. Findings The biomarker risk score had a sensitivity and specificity of 0.80 and 0.91, respectively and an AUC of 0.91 in detecting AD. When age, gender, education, and APOE status were added to the algorithm, the sensitivity, specificity, and AUC were 0.94, 0.84, and 0.95, respectively. Interpretation These initial data suggest that serum protein-based biomarkers can be combined with clinical information to accurately classify AD. Of note, a disproportionate number of inflammatory and vascular markers were weighted most heavily in analyses. Additionally, these markers consistently distinguished cases from controls in SAM, logistic regression and Wilcoxon analyses, suggesting the existence of an inflammatory-related endophenotype of AD that may provide targeted therapeutic opportunities for this subset of patients.

O'Bryant, Sid E.; Xiao, Guanghua; Barber, Robert; Reisch, Joan; Doody, Rachelle; Fairchild, Thomas; Adams, Perrie; Waring, Steven; Diaz-Arrastia, Ramon



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.



Serum levels of advanced oxidation protein products in response to allergen exposure in allergic rhinitis.  


Patients with asthma, allergic rhinitis, or atopic dermatitis experience increased oxidative stress. We conducted a prospective study to examine the levels of advanced oxidation protein products (AOPPs) as an indicator of oxidative stress in 97 patients with allergic rhinitis who were followed in our clinic during a 3.5-month period. Of these 97 patients, 51 were treated with subcutaneous immunotherapy (SCIT), and 46 did not receive any treatment until the study was concluded. In each patient, allergic rhinitis and allergic sensitization were documented by the history, the findings on clinical examination, and the results of blood and skin-prick tests. Blood samples from each patient were analyzed to determine AOPP levels. We found that the mean serum AOPP level was significantly higher in the SCIT group than in the non-SCIT group (258.55 vs. 163.83 µmol/L; p = 0.0015). We conclude that as a known indicator of protein oxidation, the serum AOPP level is a marker of increased oxidative stress in response to allergen exposure in allergic rhinitis. PMID:22930093

Aksoy, Fadlullah; Y?ld?r?m, Yavuz Selim; Veyseller, Bayram; Demirhan, Hasan; Ozturan, Orhan



Enantioselective quenching of room-temperature phosphorescence lifetimes of proteins: bovine and human serum albumins.  


Enantioselective quenching of the room-temperature phosphorescence (RTP) lifetime of proteins was demonstrated due to the effects of various external chiral quenching agents. In the absence of quenchers, the RTP lifetimes for bovine serum albumin (BSA) and human serum albumin (HSA) were found to be 5.0 +/- 0.2 and 4.0 +/- 0.1 ms, respectively. The addition of various chiral quenchers (three pairs of binaphthols and two pairs of beta-blockers) into the deoxygenated sample solutions containing BSA and HSA reduced their RTP lifetimes significantly, i.e., from ca. 4-5 ms (in the absence) to an average lifetime of ca. 1-2 ms (in the presence) of the chiral quenchers. For the R and S enantiomers examined, marked differences in RTP lifetimes were observed, i.e., ranging from ca. 20-29% for the binaphthols to ca.14-16% for the beta-blockers. Such findings could lead to a better understanding of the relationship between chirality, dynamics/conformational changes, and biological functions of proteins. PMID:17274655

Wei, Yanli; Dong, Chuan; Liu, Diansheng; Shuang, Shaomin; Huie, Carmen W



The association of c-reactive protein, serum amyloid a and fibrinogen with prevalent coronary heart disease — baseline findings of the PAIS project  

Microsoft Academic Search

Recent data suggest that infections, inflammation and the immune system are involved in the process of atherosclerosis. The aim of the present study was to analyze the association of coronary heart disease (CHD) with three inflammation markers, C-reactive protein (CRP), serum amyloid-A (SAA) and plasma fibrinogen. The cross-sectional study included 1400 men aged 45–74 years, who participated in a cardiovascular

Pekka Jousilahti; Veikko Salomaa; Vesa Rasi; Elina Vahtera; Timo Palosuo



The association of c-reactive protein, serum amyloid a and fibrinogen with prevalent coronary heart disease — baseline findings of the PAIS project  

Microsoft Academic Search

Recent data suggest that infections, inflammation and the immune system are involved in the process of atherosclerosis. The aim of the present study was to analyze the association of coronary heart disease (CHD) with three inflammation markers, C-reactive protein (CRP), serum amyloid-A (SAA) and plasma fibrinogen. The cross-sectional study included 1400 men aged 45-74 years, who participated in a cardiovascular

Pekka Jousilahti; Veikko Salomaa; Vesa Rasi; Elina Vahtera; Timo Palosuo



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.



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


The objectives of this study were to identify and compare the composition, flavor, and volatile components 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 serum and traditional whey protein manufactured from the same lot of milk. At each replication, SPC and WPC were spray dried (SD) and freeze dried (FD) to determine the effect of the heat used in spray drying on sensory properties. A trained sensory panel documented the sensory profiles of rehydrated SD or FD powders. Volatile components were extracted by solid-phase microextraction (SPME) and solvent extraction followed by solvent-assisted flavor evaporation (SAFE) with gas chromatography-mass spectrometry and gas chromatography-olfactometry. Whey protein concentrates had higher fat content, calcium, and glycomacropeptide content than SPC. Color differences (Hunter L, a, b) were not evident between SPC and WPC powders, but when rehydrated, SPC solutions were clear, whereas WPC solutions were cloudy. No consistent differences were documented in sensory profiles of SD and FD SPC and WPC. The SD WPC had low but distinct buttery (diacetyl) and cardboard flavors, whereas the SD SPC did not. Sensory profiles of both rehydrated SD products were bland and lower in overall aroma and cardboard flavor compared with the commercial WPC. Twenty-nine aroma impact compounds were identified in the SPC and WPC. Lipid and protein oxidation products were present in both products. The SPC and WPC manufactured in this study had lower total volatiles and lower concentrations of many lipid oxidation compounds when compared with commercial WPC. Our results suggest that when SPC and WPC are manufactured under controlled conditions in a similar manner from the same milk using the same ultrafiltration equipment, there are few sensory differences but distinct compositional and physical property differences that may influence functionality. Furthermore, flavor (sensory and instrumental) properties of both pilot-scale manufactured protein powders were different from commercial powders suggesting the role of other influencing factors (e.g., milk supply, processing equipment, sanitation). PMID:19762792

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



Differentially Expressed RNA from Public Microarray Data Identifies Serum Protein Biomarkers for Cross-Organ Transplant Rejection and Other Conditions  

PubMed Central

Serum proteins are routinely used to diagnose diseases, but are hard to find due to low sensitivity in screening the serum proteome. Public repositories of microarray data, such as the Gene Expression Omnibus (GEO), contain RNA expression profiles for more than 16,000 biological conditions, covering more than 30% of United States mortality. We hypothesized that genes coding for serum- and urine-detectable proteins, and showing differential expression of RNA in disease-damaged tissues would make ideal diagnostic protein biomarkers for those diseases. We showed that predicted protein biomarkers are significantly enriched for known diagnostic protein biomarkers in 22 diseases, with enrichment significantly higher in diseases for which at least three datasets are available. We then used this strategy to search for new biomarkers indicating acute rejection (AR) across different types of transplanted solid organs. We integrated three biopsy-based microarray studies of AR from pediatric renal, adult renal and adult cardiac transplantation and identified 45 genes upregulated in all three. From this set, we chose 10 proteins for serum ELISA assays in 39 renal transplant patients, and discovered three that were significantly higher in AR. Interestingly, all three proteins were also significantly higher during AR in the 63 cardiac transplant recipients studied. Our best marker, serum PECAM1, identified renal AR with 89% sensitivity and 75% specificity, and also showed increased expression in AR by immunohistochemistry in renal, hepatic and cardiac transplant biopsies. Our results demonstrate that integrating gene expression microarray measurements from disease samples and even publicly-available data sets can be a powerful, fast, and cost-effective strategy for the discovery of new diagnostic serum protein biomarkers.

Chen, Rong; Sigdel, Tara K.; Li, Li; Kambham, Neeraja; Dudley, Joel T.; Hsieh, Szu-chuan; Klassen, R. Bryan; Chen, Amery; Caohuu, Tuyen; Morgan, Alexander A.; Valantine, Hannah A.; Khush, Kiran K.; Sarwal, Minnie M.; Butte, Atul J.



Dietary casein and soybean protein affect the concentrations of serum cholesterol, triglyceride and free amino acids in rats.  


This work was undertaken to investigate the concentrations of free amino acids in blood after food was withheld from growing, male Wistar rats fed cholesterol-free, low fat (1 g corn oil/100 g) diets with casein or soybean protein for 2 wk. A diet containing 22.5 or 23.5 g/100 g of soybean protein was hypocholesterolemic compared with a diet containing 20.0 g casein/100 g diet. A comparison of serum amino acids in soybean protein-fed vs. casein-fed rats showed that, whereas concentrations of many amino acids were lower in the soybean protein-fed rats compared with the casein-fed groups, glycine was the only amino acid having a higher concentration. Further, alanine was significantly lower in the soybean protein-fed rats compared with the casein-fed rats, and the protein-induced differences in glycine and alanine concentrations of unfed rats were reproducible. When diets containing 15.0% casein or 30.0% soybean protein, a casein diet supplemented with glycine and a soybean protein diet supplemented with methionine were compared, the changes in serum glycine and alanine correlated with the changes in serum cholesterol. Concentrations of several amino acids, particularly valine, leucine and tyrosine, also changed when serum cholesterol concentrations varied, but these effects could not be explained by our experiments. The results suggest that a change in serum concentration of glycine and alanine of unfed rats may be related to the change in serum cholesterol concentration. PMID:1432265

Horigome, T; Cho, Y S



Eosinophilic myocarditis associated with dense deposits of eosinophil cationic protein (ECP) in endomyocardium with high serum ECP  

PubMed Central

A case of eosinophilic myocarditis following high serum levels of eosinophil cationic protein (ECP) is described. A 27 year old woman was admitted with New York Heart Association (NYHA) class III congestive heart failure. A haematological study showed hypereosinophilia with degranulation and vacuoles; the total eosinophil count was 7980/ml and the ECP serum concentration was noticeably high at 150 ng/ml. Endomyocardial biopsy from the right ventricle showed infiltration of eosinophils and dense deposits of ECP in the endocardium as well as the myocardium. Steroid treatment returned the total eosinophil count and serum ECP to normal, with satisfactory improvement in clinical features. Eosinophilia may cause cardiac damage, and this report confirms that eosinophil degranulation is toxic. Thus, serum ECP seems to be a reliable indicator for diagnosis and for determining treatment parameters of eosinophilic myocarditis.???Keywords: eosinophilic myocarditis; eosinophilia; eosinophil cationic protein; endomyocardial biopsy

Arima, M; Kanoh, T



Clinical significance of serum insulin-like growth factor-1 (IGF-1) and insulinlike growth factor binding protein-3 (IGFBP-3) in patients with epithelial ovarian cancer.  


Insulin-like growth factor-1 (IGF-1) and its primary binding protein IGFBP-3 play an important role in cellular proliferation, differentiation, and apoptosis in many tumors, including ovarian cancer. The objective of this study was to determine the clinical significance of the serum levels of IGF-1 and IGFBP-3 in epithelial ovarian cancer (EOC) patients. A total of 50 patients with a pathologically confirmed diagnosis of EOC were enrolled into this study. Serum IGF-1 and IGFBP-3 levels were determined by the solid-phase sandwich ELISA method. Twenty age- and sex-matched healthy controls were included in the analysis. Median age of patients was 56.5 years old (range 22 to 83 years). Majority of the patients had advanced disease (FIGO stage III-IV; 90%). No significant difference was observed in baseline serum IGF-1 and IGFBP-3 levels between EOC patients and healthy controls (p = 0.99 and p = 0.80, respectively). The young patients had higher serum IGF-1 and IGFBP-3 concentrations (p = 0.04 and p = 0.02, respectively). Patients with normal CA-125 levels had higher serum IGFBP-3 concentrations compared with those with higher CA-125 levels (p = 0.008). However, no other clinical variables including histology, tumor grade, stage of disease, and response to chemotherapy were found to be correlated with serum IGF assays (p > 0.05). A trend to significant relationship was found between the serum levels of IGF-1 and IGFBP-3 (r(s) = 0.212, p = 0.07). The patients with elevated serum IGF-1 levels had favorable progression-free and overall survivals than those with lower levels (p = 0.04 and p = 0.03, respectively). However, serum IGFBP-3 concentrations were found to have no prognostic role for both survivals (p = 0.12 and p = 0.26, respectively). In conclusion, elevated serum level of IGF-1 is associated with favorable progression-free and overall survivals in EOC patients. PMID:24254307

Tas, Faruk; Karabulut, Senem; Serilmez, Murat; Ciftci, Rumeysa; Duranyildiz, Derya



The serum protein ?2-Heremans-Schmid glycoprotein/fetuin-A is a systemically acting inhibitor of ectopic calcification  

PubMed Central

Ectopic calcification is a frequent complication of many degenerative diseases. Here we identify the serum protein ?2–Heremans-Schmid glycoprotein (Ahsg, also known as fetuin-A) as an important inhibitor of ectopic calcification acting on the systemic level. Ahsg-deficient mice are phenotypically normal, but develop severe calcification of various organs on a mineral and vitamin D–rich diet and on a normal diet when the deficiency is combined with a DBA/2 genetic background. This phenotype is not associated with apparent changes in calcium and phosphate homeostasis, but with a decreased inhibitory activity of the Ahsg-deficient extracellular fluid on mineral formation. The same underlying principle may contribute to many calcifying disorders including calciphylaxis, a syndrome of severe systemic calcification in patients with chronic renal failure. Taken together, our data demonstrate a critical role of Ahsg as an inhibitor of unwanted mineralization and provide a novel therapeutic concept to prevent ectopic calcification accompanying various diseases.

Schafer, Cora; Heiss, Alexander; Schwarz, Anke; Westenfeld, Ralf; Ketteler, Markus; Floege, Jurgen; Muller-Esterl, Werner; Schinke, Thorsten; Jahnen-Dechent, Willi



[Distribution of the ABO blood groups and the HP, TF, GC, PI and C3 serum proteins in Yakuts].  


In Yakut populations examined, polymorphisms of immunological and serum protein markers, including AB0 and Rhesus blood groups, HP, TF, GC, PI and C3, were revealed. Gene frequencies for the systems studied fell into the following ranges: AB0 system: r, 0.514 to 0.663; p, 0.136 to 0.306; q, 0.110 to 0.337; haptoglobin HP*1: 0.214 to 0.431; transferrin TF*C: 0.700 to 1.0; group specific component GC*1: 0.821 to 0.978; PI*M1 proteinase inhibitor (or alpha 1-antitrypsin) PIM1: 0.860 to 0.946; and third component of the complement C3*F: 0.031 to 0.143. PMID:12068551

Tarskaia, L A; Bychkovskaia, L S; Pa?, G V; Makarov, S V; Pakendorf, B; Spitsyn, V A



Interaction between dietary proteins and lipids in the regulation of serum and liver lipids in the rabbit. Effect of fish protein.  


Purified diets varying in dietary protein, namely casein (CA), soy protein (SP), fish protein (FP), and lipid origin (corn oil (CN), coconut oil (CO)) were fed to rabbits to evaluate the effects of protein and fat source, as well as protein-lipid interactions, on serum total, lipoprotein and hepatic lipid levels. Dietary proteins and lipids exerted a separate effect on serum total cholesterol (C), very low-density lipoprotein cholesterol (VLDL-C), and low-density lipoprotein cholesterol to high density lipoprotein cholesterol (LDL-C/HDL-C) ratio. Hence, CA increased serum cholesterol compared to SP, while coconut oil enhanced serum and VLDL-C, and decreased LDL-C/HDL-C compared to corn oil. Dietary proteins interacted with dietary lipids to modulate HDL-C levels. Thus, FP maintained a high level of HDL-C regardless of lipid origin, compared to CA and SP whose HDL-C levels were decreased by corn oil, compared to coconut oil. A dietary protein-lipid interaction was also observed in the regulation of liver cholesterol levels. Coconut oil, compared to corn oil, decreased liver cholesterol in rabbits fed FP, whereas hepatic cholesterol concentration was unaltered by dietary lipid source in CA- and SP-fed rabbits. These results demonstrate that dietary proteins act synergistically with dietary lipids to regulate cholesterol metabolism in the rabbit. PMID:1762524

Bergeron, N; Deshaies, Y; Lavigne, C; Jacques, H



Protein interactions with polyelectrolyte multilayers: interactions between human serum albumin and polystyrene sulfonate/polyallylamine multilayers.  


The interactions between polystyrenesulfonate (PSS)/polyallylamine (PAH) multilayers with human serum albumin (HSA) were investigated by means of scanning angle reflectometry (SAR). We find that albumin adsorbs both on multilayers terminating with PSS (negatively charged) or PAH (positively charged) polyelectrolytes. On films terminating with PSS only, an albumin equivalent monolayer is found whereas when PAH constitutes the outer layer, albumin interacts with the multilayer in such a way as to form a protein film that extends over thicknesses that can be as high as four times the largest dimension of the native albumin molecule. Once the protein film is formed, it is found that when the albumin solution is replaced by a pure buffer solution of same ionic strength as the adsorption solution almost no desorption takes place. On the other hand, when a buffer solution of higher ionic strength is brought in contact with the albumin film, a significant amount of adsorbed proteins is released. One also observes that, for albumin solutions of a given protein concentration, the adsorbed protein amount depends on the ionic strength of the adsorption solution. On surfaces terminating with PAH, the adsorbed protein amount first increases rapidly but passes through a maximum and decreases with the ionic strength. The ionic strength corresponding to the maximum of the adsorbed albumin amount itself depends on the albumin concentration. On the other hand, on films terminating with PSS the adsorbed amount increases with the salt concentration before leveling-off. These results show that the underlying complexity of concentration and pH dependent adsorption/desorption equilibria often simply termed "protein adsorption" is the result of antagonist competing interactions that are mainly of electrostatic origin. We also propose two microscopic models, that are compatible with our experimental observations. PMID:11710198

Ladam, G; Gergely, C; Senger, B; Decher, G; Voegel, J C; Schaaf, P; Cuisinier, F J



Novel Roles of Specific Isoforms of Protein Kinase C in Activation of the c-fos Serum Response Element  

Microsoft Academic Search

Protein kinase C (PKC) is a multigene family of enzymes consisting of at least 11 isoforms. It has been implicated in the induction of c-fos and other immediate response genes by various mitogens. The serum response element (SRE) in the c-fos promoter is necessary and sufficient for induction of transcription of c-fos by serum, growth factors, and the phorbol ester



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

Microsoft Academic Search

\\u000a Increased understanding of influential factors for the cultivation of animal cells, combined with heightened regulatory concern\\u000a over potential transmission of adventitious contaminants associated with serum and other animal-derived components, has elevated\\u000a interest in using protein hydrolysates as serum replacements or nutrient supplements. This paper reviews the chemistry and\\u000a biology of various hydrolysates derived from animal, plant and microbial sources. It

Juliet Lobo-Alfonso; Paul Price; David Jayme



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


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. PMID:24976158

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



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

PubMed Central

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



C-reactive protein concentrations in serum of dogs with naturally occurring renal disease.  


The current study was undertaken to investigate the relation between serum C-reactive protein (CRP) concentrations and parameters of renal function in dogs with naturally occurring renal disease. Dogs were assigned to groups according to plasma creatinine concentration, urinary protein-to-creatinine ratio (UP/UC), and exogenous plasma creatinine clearance (P-Cl(Cr)) rates. Group A (healthy control dogs; n = 8): non-azotemic (plasma creatinine <125 µmol/l) and nonproteinuric (UP/UC <0.2), with P-Cl(Cr) rates >90 ml/min/m(2); group B (n = 11): non-azotemic, nonproteinuric dogs with reduced P-Cl(Cr) rates (50-89 ml/min/m(2)); group C (n = 7): azotemic, borderline proteinuric dogs (P-Cl(Cr) rates: 22-67 ml/min/m(2)); and group D (n = 6): uremic, proteinuric dogs (not tested for P-Cl(Cr)). The serum CRP concentrations were measured via commercial enzyme-linked immunosorbent assay. The CRP concentrations in the clinically healthy dogs (group A) ranged from 2.09 mg/l to 8.60 mg/l (median: 3.21 mg/l). In comparison with dogs of group A, median CRP concentrations were significantly (P < 0.01) elevated in dogs of group B (17.6 mg/l, range: 17.0-19.2 mg/l), group C (24.8 mg/l, range: 18.0-32.5 mg/l), and group D (59.7 mg/l, range: 17.7-123 mg/l). Serum CRP was significantly related to P-Cl(Cr) (r = -0.83; P < 0.001), plasma creatinine (r = 0.81; P < 0.001), UP/UC (r = 0.70; P < 0.001), and leukocytes (r = 0.49; P < 0.01). The significant relations between serum CRP concentrations and biochemical parameters of kidney function in plasma and urine suggest that a stimulation of the acute phase response is implicated in the pathogenesis of canine renal disease. PMID:21908313

Raila, Jens; Schweigert, Florian J; Kohn, Barbara



[Does the assay of acute phase protein concentrations in cerebrospinal fluid and/or in serum in patient with viral meningitis have a diagnostic value? Part II. Lymphocytic meningitis caused by echo 30 virus].  


The aim of the study was to evaluate dynamics of selected acute phase proteins in serum and cerebrospinal fluid (CSF) in children with viral meningitis and to assess diagnostic power of protein determination for detection and treatment monitoring. 51 children with viral meningitis caused by ECHO 30 virus were included in the study group. Concentration of C-reactive protein (CRP), alpha 1-antitrypsin (AAT), alpha1-acid glycoprotein (AAG), alpha2-haptoglobin (HPT) and C3 complement fragment were determined in serum and CSF at entry and at day 14 after admittance to hospital. Control group for serum determination consisted in 30 healthy children (Group K1) and control group for CSF determination consisted in 19 hospitalized children in whom the diagnosis of meningitis was not confirmed (group K2). The greatest rise of acute phase proteins concentration was observed in children in case of HPT, AAG and C3 complement when determined in serum. Meningitis in children that was caused by ECHO 30 virus produces slight acute phase reaction that is more evident in serum than in CSF. It is confirmed by remarkable increase of AAG, HPT, C3 complement in serum and HPT in CSF either at entry or at the day 14. The determination of AAG, HPT and C3 complement in serum have diagnostic power that is strong enough to meningitis diagnostics and monitoring of treatment. PMID:15517816

Mame?ka, Beata; Lobos, Marek; Sass-Just, Maria; Dworniak, Daniela; Urbaniak, Anna; Terlecka, Monika; Paradowski, Marek



Redox and label-free array detection of protein markers in human serum.  


A substantial outstanding challenge in diagnostics and disease monitoring is an ability to rapidly and conveniently assay for protein biomarkers within complex biological media. Label-free electroanalytical methods present, arguably, the most promising and scalable means of achieving this but, as with all label-free assays, can struggle with response selectivity issues that arise from nonspecific surface interactions. Impedimetric methods are ultrasensitive and have been applied to the quantification of a wide range of proteins but have not previously been utilized in a multiplexed format capable of operation in complex analytical fluid. Herein, we present the use of thermally cross-linked poly(ethylene glycol) (PEG) polymer sensory array interfaces in the ultrasensitive quantification of two protein markers, insulin and C-reactive protein (CRP). This was achieved with detection limits of 171 ± 19 fM and 150 ± 10 pM, respectively. Significantly, the arrays not only enable the simultaneous, fast, nonamplified, and label-free detection of both markers without reagent addition but do so with little cross talk, even in human serum. A blind analysis of 17 real patient samples generated results in excellent agreement with those obtained through a clinically approved chemiluminescence assay. PMID:24813814

Luo, Xiliang; Xu, Qiao; James, Tim; Davis, Jason J



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

PubMed Central

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

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



First evidence of protein G-binding protein in the most primitive vertebrate: serum lectin from lamprey (Lampetra japonica).  


The intelectins, a recently identified subgroup of extracellular animal lectins, are glycan-binding receptors that recognize glycan epitopes on foreign pathogens in host systems. Here, we have described NPGBP (novel protein G-binding protein), a novel serum lectin found in the lamprey, Lampetra japonica. RT-PCR yielded a 1005 bp cDNA sequence from the lamprey liver encoding a 334 amino acid secretory protein with homology to mammalian and aquatic organism intelectins. Gene expression analyses showed that the NPGBP gene was expressed in the blood, intestines, kidney, heart, gill, liver, adipose tissue and gonads. NPGBP was isolated by protein G-conjugated agarose immunoprecipitation, and SDS-PAGE analyses showed that NPGBP migrated as a specific band (?35 and ?124 kDa under reducing and non-reducing conditions, respectively). These results suggested that NPGBP forms monomers and tetramers. NPGBP gene expression was induced by in vivo bacterial stimulation, and NPGBP showed different agglutination activities against pathogenic Gram-positive bacteria, Gram-negative bacteria and fungi. The induction of NPGBP suggested that it plays an important role in defense against microorganisms in the internal circulation system of the lamprey. When incubated with an unrelated antibody, the specific binding between NPGBP and protein G was competitively inhibited, indicating that NPGBP and the Fc region of Ig bind to the same site on protein G. We thus assume that the tertiary structure of NPGBP is similar to that of the Fc region of Ig. Additionally, NPGBP can effectively promote endothelial cell mitosis. These findings suggest that NPGBP plays a role in the immune defense against microorganisms, and this study represents one of the few examples of the characterization and functional analysis of an aquatic organism intelectin. PMID:23806362

Xue, Zhuang; Pang, Yue; Liu, Xin; Zheng, Zhen; Xiao, Rong; Jin, Minli; Han, Yinglun; Su, Peng; Lv, Li; Wang, Jihong; Li, QingWei



Methodological Challenges of Protein Analysis in Blood Serum and Cerebrospinal Fluid by Capillary Electrophoresis  

Microsoft Academic Search

Summary High-performance capillary electrophoresis (HPCE or CE) is an ultrasensitive analytical technique with high resolving power and a wide area of applications including peptide\\/protein analysis. Its applicability is greatly enhanced by the short separation times, the ease of method development and the minimum sample and organic solvent requirements. Various HPCE modes have been developed for peptide\\/protein analysis, including capillary zone

F. N. Lamari; N. K. Karamanos



Serum levels of eosinophil cationic protein, eosinophil-derived neurotoxin and myeloperoxidase in infections with filariea and schistosomes  

Microsoft Academic Search

The serum levels of three major granulocyte proteins were measured in patients with onchocerciasis, bancroftian filariasis and intestinal schistosomiasis and compared to controls from patients with malaria, Africans living in areas not endemic for these infections and healthy Germans. The investigation comprised the determination of the eosinophil granule proteins eosinophil cationic protein (ECP) and eosinophil-derived neurotoxin (EDN\\/EPX), and the neutrophil\\/monocyte

F. W. Tischendorf; N. W. Brattig; D. W. Büttner; A. Pieper; M. Lintzel



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


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

Sadeghi, Gh; Pourreza, J



ABT-737 promotes the dislocation of ER luminal proteins to the cytosol, including pseudomonas exotoxin.  


Impaired apoptosis is often a key element in tumor development. Therefore, drugs mimicking prosurvival antagonists offer promise as cancer therapeutics. When ABT-737, a BH3-only mimetic, was added to KB3-1 human cervical adenocarcinoma cells, we noted an induction of an endoplasmic reticulum (ER) stress response and the dislocation of ER luminal proteins, including chaperones, to the cell cytosol. Furthermore, when immunotoxin (antibody-toxin chimeric molecule) and ABT-737 combinations were added to cells, there was enhanced toxin-mediated inhibition of protein synthesis, consistent with enhanced translocation of toxin to the cytosol. A similar enhancement was not seen with thapsigargin, suggesting that ER stress alone was not responsible for enhanced translocation. Cytosol preparations from ABT-737-treated but not from thapsigargin-treated cells revealed the presence of greater amounts of processed 37-kDa toxin fragment compared with the addition of immunotoxin alone. As early as 4 hours after the addition of ABT-737 and immunotoxin, there was release of mitochondrial cytochrome c and activation of caspase-3/7 indicating that the combination caused apoptotic cell death. These results were reflected in decreased cellular ATP levels that were noted with combinations of ABT-737 and immunotoxin but not with either agent alone or with combinations of thapsigargin and immunotoxin. We conclude that ABT-737 increases ER permeability, promoting the dislocation of toxin from the ER to the cytosol resulting in early apoptotic cell death. These mechanistic insights suggest why this class of BH3-only mimetic synergizes in a particular way with Pseudomonas exotoxin-based immunotoxins. PMID:24739394

Antignani, Antonella; Sarnovsky, Robert; FitzGerald, David J



Comparable autoantibody serum levels against amyloid- and inflammation-associated proteins in Parkinson's disease patients and controls.  


Naturally occurring autoantibodies (NAbs) against a number of potentially disease-associated cellular proteins, including Amyloid-beta1-42 (Abeta1-42), Alpha-synuclein (Asyn), myelin basic protein (MBP), myelin oligodendrocyte glycoprotein (MOG), and S100 calcium binding protein B (S100B) have been suggested to be associated with neurodegenerative disorders, in particular Alzheimer's (AD) and Parkinson's disease (PD). Whereas the (reduced) occurrence of specific NAbs in AD is widely accepted, previous literature examining the relation of these NAb titres between PD patients and controls, as well as comparing these levels with demographic and clinical parameters in PD patients have produced inconsistent findings. We therefore aimed, in a cross-sectional approach, to determine serum titres of the above NAbs in a cohort of 93 PD patients (31 of them demented) and 194 controls. Levels were correlated with demographic and clinical variables, cerebrospinal fluid Abeta1-42, total tau and phospho-tau levels, as well as with single nucleotide polymorphisms (SNPs) of genes which either have been reported to influence the immune system, the amyloid cascade or the occurrence of PD (ApoE, GSK3B, HLA-DRA, HSPA5, SNCA, and STK39). The investigated NAb titres were neither significantly associated with the occurrence of PD, nor with demographic and clinical parameters, neurodegenerative markers or genetic variables. These results argue against a major potential of blood-borne parameters of the adaptive immune system to serve as trait or state markers in PD. PMID:24586351

Maetzler, Walter; Apel, Anja; Langkamp, Markus; Deuschle, Christian; Dilger, Sarah Selina; Stirnkorb, Johannes Georg; Schulte, Claudia; Schleicher, Erwin; Gasser, Thomas; Berg, Daniela



The Cellulosome System of Acetivibrio cellulolyticus Includes a Novel Type of Adaptor Protein and a Cell Surface Anchoring Protein  

PubMed Central

A scaffoldin gene cluster was identified in the mesophilic cellulolytic anaerobe Acetivibrio cellulolyticus. The previously described scaffoldin gene, cipV, encodes an N-terminal family 9 glycoside hydrolase, a family 3b cellulose-binding domain, seven cohesin domains, and a C-terminal dockerin. The gene immediately downstream of cipV was sequenced and designated scaB. The protein encoded by this gene has 942 amino acid residues and a calculated molecular weight of 100,358 and includes an N-terminal signal peptide, four type II cohesions, and a C-terminal dockerin. ScaB cohesins 1 and 2 are very closely linked. Similar, but not identical, 39-residue Thr-rich linker segments separate cohesin 2 from cohesin 3 and cohesin 3 from cohesin 4, and an 84-residue Thr-rich linker connects the fourth cohesin to a C-terminal dockerin. The scaC gene downstream of scaB codes for a 1,237-residue polypeptide that includes a signal peptide, three cohesins, and a C-terminal S-layer homology (SLH) module. A long, ca. 550-residue linker separates the third cohesin and the SLH module of ScaC and is characterized by an 18-residue Pro-Thr-Ala-Ser-rich segment that is repeated 27 times. The calculated molecular weight of the mature ScaC polypeptide (excluding the signal peptide) is 124,162. The presence of the cohesins and the conserved SLH module implies that ScaC acts as an anchoring protein. The ScaC cohesins are on a separate branch of the phylogenetic tree that is close to, but distinct from, the type I cohesins. Affinity blotting with representative recombinant probes revealed the following specific intermodular interactions: (i) an expressed CipV cohesin binds selectively to an enzyme-borne dockerin, (ii) a representative ScaB cohesin binds to the CipV band of the cell-free supernatant fraction, and (iii) a ScaC cohesin binds to the ScaB dockerin. The experimental evidence thus indicates that CipV acts as a primary (enzyme-recognizing) scaffoldin, and the protein was also designated ScaA. In addition, ScaB is thought to assume the role of an adaptor protein, which connects the primary scaffoldin (ScaA) to the cohesin-containing anchoring scaffoldin (ScaC). The cellulosome system of A. cellulolyticus thus appears to exhibit a special type of organization that reflects the function of the ScaB adaptor protein. The intercalation of three multiple cohesin-containing scaffoldins results in marked amplification of the number of enzyme subunits per cellulosome unit. At least 96 enzymes can apparently be incorporated into an individual A. cellulolyticus cellulosome. The role of such amplified enzyme incorporation and the resultant proximity of the enzymes within the cellulosome complex presumably contribute to the enhanced synergistic action and overall efficient digestion of recalcitrant forms of cellulose. Comparison of the emerging organization of the A. cellulolyticus cellulosome with the organizations in other cellulolytic bacteria revealed the diversity of the supramolecular architecture.

Xu, Qi; Gao, Wenchen; Ding, Shi-You; Kenig, Rina; Shoham, Yuval; Bayer, Edward A.; Lamed, Raphael



Role of serum procalcitonin and C-reactive protein in the diagnosis of neonatal sepsis.  


This cross sectional observational study was done in the division of neonatology, department of pediatrics, Bangabandhu Sheikh Mujib Medical University (BSMMU) in the year 2007. The study population was 50 newborns in total who needed evaluation of sepsis on clinical suspicion. The main objective of this study was to assess serum procalcitonin (PCT) as a better diagnostic marker than C-Reactive Protein (CRP) in neonatal sepsis. The total study populations were classified into 4 groups like highly probable, probable, and possible and no sepsis group according to the clinical and blood parameters. PCT and CRP were assessed and compared by statistical analysis. For the estimation of PCT and CRP, venous blood was drawn and centrifuged and stored at - 20 degrees C in the refrigerator. Later on PCT was measured by rapid semi quantitative immunochromatographic test. Level of CRP was determined by semi quantitative method (latex). All data were analyzed by SPSS version 10 windows. For statistical analysis appropriate tests were done. In all observations sepsis was found to be more common in male newborns and in those who were delivered by caesarean section. In low birth weight and preterm newborns sepsis was more prevalent. Premature rupture of membrane (PROM) was found to be the commonest maternal clinical condition as a risk factor of sepsis. There was positive correlation between serum PCT and CRP and values of serum PCT as well as CRP differed significantly in the different categories of sepsis indicating relation to the severity of sepsis. PCT is a useful, sensitive and independent biomarker of neonatal sepsis. CRP measurement along with PCT measurement may increase the specificity. Though PCT measurement is comparatively expensive but an easy bed side promt convenient procedure for sick neonates in addition to CRP for rapid evaluation of neonatal sepsis rather than waiting for the report of blood culture. PMID:21877603

Naher, B S; Mannan, M A; Noor, K; Shahiddullah, M



Serum Cellular Apoptosis Susceptibility Protein Is a Potential Prognostic Marker for Metastatic Colorectal Cancer  

PubMed Central

Colorectal cancer has high rates of recurrence and metastasis. Many patients with similar histopathological features show significantly different clinical outcomes, and these differences are primarily related to metastases undetected by current diagnostic methods. There is no useful serological marker for metastatic disease. We investigated the cellular apoptosis susceptibility (CSE1L/CAS) protein in comparison with carcinoembryonic antigen (CEA) as a marker for metastatic colorectal cancer. Using serum from 103 patients with stage I, II, III, and IV disease, CSE1L was detected in 36.0% (9 of 25), 57.7% (15 of 26), 71.4% (30 of 42), and 88.9% (8 of 9) of patients, respectively; a pathological CEA level was found in 16.0% (4 of 25), 42.3% (11 of 26), 47.6% (20 of 42), and 77.8% (7 of 9) of patients, respectively; a combined CSE1L/CEA assay was detected in 48.0% (12 of 25), 65.4% (17 of 26), 88.1% (37 of 42), and 100% (9 of 9) of patients, respectively. Lymphatic metastasis is an important predictor of poor prognosis and crucial for determination of therapeutic strategy. Serum CSE1L was detected in 74.5% (38 of 51) of patients with lymph node metastasis, whereas a pathological CEA level was found in only 52.9% (27 of 51) of the same patients (P < 0.001); the combined CSE1L/CEA assay increased sensitivity to 90.2% (46 of 51). Animal experiments showed CSE1L reduction in B16-F10 melanoma cells correlated with decreased metastasis to the colorectal tract in C57BL/6 mice. These results indicate that assay of serum CSE1L may facilitate diagnosis of colorectal cancer lymphatic metastases; furthermore, CSE1L is a possible therapeutic target.

Stella Tsai, Chin-Shaw; Chen, Hung-Chang; Tung, Jai-Nien; Tsou, Shung-Sheng; Tsao, Tang-Yi; Liao, Ching-Fong; Chen, Ying-Chun; Yeh, Chi-Yuan; Yeh, Kun-Tu; Jiang, Ming-Chung



Enrichment of serum low-molecular-weight proteins using C18 absorbent under urea/dithiothreitol denatured environment.  


Serum low-molecular-weight proteins (LMWPs, molecular weight<30kDa) are closely related to the body physiological and pathological situations, whereas many difficulties are encountered when enriching and fractionating them. Using C(18) absorbent (100 A) enrichment and fractionation under urea/dithiothreitol (DTT) denatured environment followed by 60% acetonitrile (ACN) elution, serum LMWPs could be enriched more than 100-fold and were evaluated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), two-dimensional gel electrophoresis (2-DE), and isotope-coded affinity tag (ICAT) labeling quantification. Proteins existing in human serum at low nanograms/milliliter (ng/ml) levels, such as myeloid-related proteins (MRPs), could be identified directly from 2-DE coupled with matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry (MALDI-TOF/TOF MS) and LTQ-Orbitrap MS. Sixteen proteins were confidentially identified and quantified using ICAT labeling and liquid chromatography-tandem mass spectrometry (LC-MS/MS). By virtue of its easy operation and high reproducibility to process large quantity complex serum samples, this method has potential uses in enriching LMWPs either in serum or in cell and tissue samples. PMID:19891953

Wu, Jing; An, Yuan; Pu, Hai; Shan, Yue; Ren, Xiaoqing; An, Mingrui; Wang, Qingsong; Wei, Shicheng; Ji, Jianguo



In vitro serum protein-binding characteristics of bis-(2-ethylhexyl) phthalate and its principal metabolite, mono-(2-ethylhexyl) phthalate  

SciTech Connect

The metabolism and toxicity of the ubiquitous plasticizer, bis-(2-ethylhexyl) phthalate (DEHP), and its principal metabolite, mono-(2-ethylhexyl) phthalate (MEHP), have been extensively investigated. In an attempt to understand their disposition in man, the authors studied the in vitro serum protein-binding characteristics of these compounds, using ultracentrifugation and agarose gel electrophoresis. The association of DEHP and lipoproteins was shown to be highly dependent upon, and proportional to, the lipid concentration of the serum. It appears that more than half of the serum DEHP is bound to proteins with density greater than 1.21 g/mL when the concentration of cholesterol is below 300 mg/dL or the cholesterol and triglyceride total concentration is less than 600 mg/dL. As the cholesterol and triglyceride concentrations increase, the percent DEHP bound to VLDL, IDL, and LDL increases. MEHP is bound principally to nonlipoprotein constituents in the serum, and this binding distribution is unaffected by lipid concentration. The percent binding of DEHP and MEHP to individual proteins was also found to be unaffected by their concentrations in serum. These data indicate that the protein-binding characteristics of these compounds, in vitro, is somewhat more complex than previously reported.

Griffiths, W.C.; Camara, P.D.; Saritelli, A.; Gentile, J.



Adiponectin, chemerin, cytokines, and dipeptidyl peptidase 4 are released from human adipose tissue in a depot-dependent manner: an in vitro system including human serum albumin  

PubMed Central

Background Adipose tissue (AT) contributes to metabolic dysfunction through imbalanced production of adipokines, including cytokines. Visceral AT in particular is associated with metabolic disorders, indicating a specific secretory status. The relative significance of different human AT depots in adipokine release is not fully known. Further, previous in vitro systems usually included medium containing bovine serum albumin (BSA), which may induce cytokine release. Our aim was to compare release of a number of adipokines/cytokines – all implicated in insulin resistance – from human subcutaneous and visceral AT in a short-term incubation system minimizing cytokine induction and including repeated measurements during 24 h. A prerequisite was to evaluate a potential alternative to BSA in the incubation medium. Methods Subcutaneous and/or visceral AT from 17 patients (age 20–68 years; BMI 22.6–56.7 kg/m2) undergoing elective surgery was incubated for 2, 4, 6, 8, and 24 h in medium with or without 1% BSA or human serum albumin (HSA). Medium concentrations of adiponectin, chemerin, nine cytokines, dipeptidyl peptidase 4 (DPP4), and omentin were analyzed by multiplex immunoassay or ELISA. Adipocyte size, AT macrophage density, and medium concentrations of endotoxin were determined. Results Cytokine release was induced by BSA but not by HSA. In evaluation of the final incubation protocol including 1% HSA, and as expected, adiponectin release was higher from subcutaneous biopsies of nonobese than of obese subjects and inversely associated with adipocyte size; omentin was released almost exclusively from visceral AT. Exploratory incubations revealed more abundant release of chemerin, cytokines (except IL-6), and DPP4 from the visceral depot, while adiponectin release was higher from subcutaneous than visceral AT. Release was linear for a maximum of 2–6 h. Macrophage density was higher in visceral than subcutaneous AT. Levels of endotoxin in the medium were negligible. Conclusions Adiponectin, chemerin, many cytokines, and DPP4 are released from human AT in a depot-dependent manner. These results highlight functional differences between visceral and subcutaneous AT, and a mechanistic link between regional fat accumulation and metabolic disorders. Supplementation of human AT incubation medium with HSA rather than BSA is recommended to minimize induction of cytokine release.



Changes of Serum Retinol Binding Protein 4 Levels Following 8 Weeks Moderate Aerobic Exercise  

PubMed Central

Purpose The purpose of this study was to examine the effects of 8 weeks moderate intensity aerobic exercise on serum retinol binding protein 4 (RBP4) levels in female athletes. Methods Twenty female karate athletics were randomly assigned to one of the exercise group (n = 10) or control group (n = 10). The training group performed endurance training 3 days a week for 8 weeks at an intensity corresponding to 50-60% individual maximum oxygen consumption for 45 min. Results Body mass and body mass index increased (P < 0.05) after 8 weeks aerobic exercise compared to the control group. For waist to hip ratio (WHR), body fat percentage and maximal oxygen consumption there were no significant differences between the exercise group and the control group. There were virtually no changes in body fat percentage, fasting glucose, insulin, insulin resistance and RBP4 levels after 8 weeks training. Conclusion Serum RBP4 level was not affected by 8-week moderate aerobic exercise in female athletes.

Ahmadi, Narges; Moghadasi, Mehrzad; Nuri, Reza



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.

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



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


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



Serum adipocyte fatty acid-binding protein is independently associated with complex coronary lesions in patients with stable coronary artery disease.  


The association between circulating adipocyte fatty acid-binding protein (A-FABP) levels and coronary artery disease (CAD) is reported. We assessed whether plasma A-FABP levels are associated with angiographic coronary lesion morphology in patients with stable CAD. Serum A-FABP levels were analyzed in 115 patients with stable CAD (mean age 69 ± 10 years; 80 % men). These patients were angiographically studied and divided into two groups: simple lesions (n = 34) and complex lesions (n = 81). We also compared 50 age- and gender-matched controls with no evidence of CAD. Serum A-FABP levels in patients with stable CAD were significantly higher than those in controls. In patients with stable CAD, serum A-FABP levels were significantly higher in patients with complex lesions than in those with simple lesions: median (25th-75th percentile), 23.4 (17.7-30.8) vs 18.2 (12.2-24.7) ng/ml, P < 0.01. Serum A-FABP levels were also significantly associated with angiographic scores of extent of coronary lesion (r = 0.21, P = 0.02). Multiple logistic analysis that included dyslipidemia, statin therapy, and extent score demonstrated that serum A-FABP was independently associated with complex lesions. The multiple adjusted odds ratio for a complex lesion with a serum A-FABP level (per doubling) was 2.38 (95 % confidence interval, 1.03-6.41; P = 0.03). High serum A-FABP levels were significantly associated with complex coronary lesions in patients with stable CAD, suggesting that high A-FABP levels may be involved in coronary plaque vulnerability. PMID:23224329

Kajiya, Masahito; Miyoshi, Toru; Doi, Masayuki; Usui, Shinichi; Iwamoto, Mutsumi; Takeda, Ko; Nosaka, Kazumasa; Nakayama, Rie; Hirohata, Satoshi; Kusachi, Shozo; Nakamura, Kazufumi; Ito, Hiroshi



Influence of Polymorphism on Glycosylation of Serum Amyloid A4 Protein  

PubMed Central

Serum amyloid A4 (SAA4) is a constitutive apolipoprotein of high-density lipoprotein. It exhibits N-linked glycosylation in its second half. There are both glycosylated and nonglycosylated forms in plasma and the ratio of these two forms varies among individuals. This study was conducted to examine the influence of genetic polymorphism of SAA4 on its glycosylation status. In 55 healthy subjects, SAA4 polymorphism was analyzed by PCR combined direct sequencing and its glycosylation status was analyzed by immunoblotting. The results showed that the percentage of glycosylation in subjects with amino acid substitutions at positions 71 and/or 84 was significantly (P < 0.05) higher than that in subjects with the wild type. The polymorphism had no influence on the plasma concentration of SAA4. These findings suggest that the changes in protein structures alter the efficiency of glycosylation in the SAA4 molecule. The functional implication of this should be of interest.

Kotani, Kazuhiko; Tanaka, Masafumi



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.

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



Further characterization of the Ca2+-dependent F-actin-depolymerizing protein of human serum.  


The F-actin-depolymerizing factor (ADF) of human serum was purified and identified as a 93000-daltons protein. The concentration of ADF was calculated to 120-150 micrograms/ml in four normal sera and about half as much in three sera from leukemia patients treated with cytostatic drugs. In the presence of Ca2+ ADF shortened actin filaments into fragments, the size of which was correlated to the actin: ADF molar ratio, as judged by electron microscopy. The severing of F-actin was not necessarily followed by an increase in the quantities of monomeric actin, as determined by a DNase I inhibition assay and a sedimentation assay. The findings indicated that ADF shortens filamentous actin by breaking bonds between adjacent actin molecules thereby forming stable ADF-actin complexes, without a monomeric net release. The effect of ADF on F-actin was rapid and was reversed upon chelation of Ca2+. ADF cross-reacted immunologically and exhibited similarity in reaction mechanism with gelsolin, the Ca2+-dependent F-actin-severing protein from macrophages. This implies that the proteins are both structurally and functionally related. The physiological role of ADF may be to handle actin released at cell destruction, probably by forming ADF-G-actin 1:1 complexes thereby preventing formation of actin filaments. PMID:7128580

Thorstensson, R; Utter, G; Norberg, R



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


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

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



A fully integrated multi-column system for abundant protein depletion from serum/plasma.  


This work details the transformation of a conventional HPLC system to a low back pressure liquid chromatography set-up for automated serum/plasma depletion and fractionation. A Dionex U3000 HPLC was converted to low back pressure operation (125 psi max) by replacing all narrow-bore lines to larger inner-diameter tubing. The system was configured to use two immunoaffinity columns, first for depletion of the top 14 most abundant proteins (Seppro IgY14), then for the next 200-300 proteins (Seppro SuperMix). The autosampler was dual-purposed for both injection and fraction collection. Both the flow-through and SuperMix bound proteins were collected in an automated fashion. Three samples could be depleted consecutively before the system required user intervention, and up to nine samples could be depleted within a 24 h period. This study documents the validation of the instrument performance with a 90-patient sample set, demonstrating overall CVs for 86 of the 90 samples to be within the 95% confidence intervals. Additionally, there was excellent reproducibility within the same patient (biological replicates) across days. PMID:22771237

Janecki, Dariusz J; Pomerantz, Steven C; Beil, Eric J; Nemeth, Jennifer F



Discrete serum protein signatures discriminate between human retrovirus-associated hematologic and neurologic disease.  


The human T-cell leukemia virus type I (HTLV-I) is the causative agent for adult T-cell leukemia (ATL) and HTLV-I-associated myelopathy/tropical spastic paraparesis (HAM/TSP). Approximately 5% of infected individuals will develop either disease and currently there are no diagnostic tools for early detection or accurate assessment of disease state. We have employed high-throughput expression profiling of serum proteins using mass spectrometry to identify protein expression patterns that can discern between disease states of HTLV-I-infected individuals. Our study group consisted of 42 ATL, 50 HAM/TSP, and 38 normal controls. Spectral peaks corresponding to peptide ions were generated from MS-TOF data. We applied Classification and Regression Tree analysis to build a decision algorithm, which achieved 77% correct classification rate across the three groups. A second cohort of 10 ATL, 10 HAM and 10 control samples was used to validate this result. Linear discriminate analysis was performed to verify and visualize class separation. Affinity and sizing chromatography coupled with tandem mass spectrometry was used to identify three peaks specifically overexpressed in ATL: an 11.7 kDa fragment of alpha trypsin inhibitor, and two contiguous fragments (19.9 and 11.9 kDa) of haproglobin-2. To the best of our knowledge, this is the first application of protein profiling to distinguish between two disease states resulting from a single infectious agent. PMID:15889159

Semmes, O J; Cazares, L H; Ward, M D; Qi, L; Moody, M; Maloney, E; Morris, J; Trosset, M W; Hisada, M; Gygi, S; Jacobson, S



Quantification of nonenzymically glycated albumin and total serum protein by affinity chromatography.  


We have evaluated an affinity-chromatographic procedure for determination of glycated albumin (GA) and glycated total serum protein (GSP). Recovery of these analytes was inversely related to free glucose concentration, thus necessitating removal of free glucose. For this we used molecular-exclusion chromatography on G-25 Sephadex, or dialysis, the latter procedure resulting in significantly (p less than 0.05) lower concentrations of GSP and GA. Total protein concentration and percent glycation are also inversely related, and so protein concentrations must be standardized before the assay. Within- and between-run CVs for both GSP and GA were less than 6.5 and 18%, respectively, the determination of GA being generally the more precise of the two. Labile glycated fractions, lipemia, icterus, hemolysis, and type of anticoagulant did not affect the results, but assay temperature did. Diabetic subjects showed substantially higher concentrations of GA and GSP than did normal subjects. Because of the life span of these analytes in circulation, their measurement may provide a short-term index of glycemic control. PMID:6697494

Yatscoff, R W; Tevaarwerk, G J; MacDonald, J C



Milk-derived proteins and minerals alter serum osteocalcin in prepubertal boys after 7 days.  


We have previously shown that at equal protein content, milk, but not meat, decreased bone turnover in boys. This suggested that milk-derived components are important for bone metabolism. In the present study, we hypothesized that milk-derived proteins (whey and casein) affect bone turnover during growth depending on the content of milk minerals (calcium and phosphorus). This was a randomized, parallel, double-blind study. Eight-year-old boys (n = 57) received 1 of 4 milk drinks: whey protein with low or high content of minerals, or casein protein with low or high content of minerals. The amount of whey and casein was identical to their content in 1.5 L of milk. We measured serum osteocalcin (sOC), bone-specific alkaline phosphatase, and C-terminal telopeptides of type I collagen (immunoassay) and estimated dietary intake (3-day weighed food record) at baseline and after 7 days. Only sOC was significantly affected by the treatments (P < .05). There was a significant interaction between milk-derived proteins and minerals with regard to sOC (P = .01). The intake of milk drinks containing whey increased sOC at the low content of minerals, whereas it decreased sOC at the high content of minerals (P < .05). In contrast, milk drinks containing casein increased sOC both at the low and at the high contents of minerals. In conclusion, whey and casein (corresponding to their content in 1.5 L of milk) differently affect sOC in 8-year-old boys depending on the content of milk minerals, but do not seem to affect other markers for bone turnover. PMID:20851310

Mark, Alicja Budek; Hoppe, Camilla; Michaelsen, Kim F; Mřlgaard, Christian



Effects of uremic toxins and fatty acids on serum protein binding of furosemide: possible mechanism of the binding defect in uremia  

Microsoft Academic Search

To elucidate the mechanism of impaired serum binding of furosemide observed in patients with renal dysfunc- tion, we examined in vitro the serum protein binding of furosemide in the absence and presence of uremic toxins that are endogenously retained solutes in uremic serum and act as inhibitors of drug binding. Analysis of the binding data of furosemide at its therapeutic

Norito Takamura; Toru Maruyama; Masaki Otagiri


Proteomics analysis of serum protein profiling in pancreatic cancer patients by DIGE: up-regulation of mannose-binding lectin 2 and myosin light chain kinase 2  

Microsoft Academic Search

BACKGROUND: Pancreatic cancer has significant morbidity and mortality worldwide. Good prognosis relies on an early diagnosis. The purpose of this study was to develop techniques for identifying cancer biomarkers in the serum of patients with pancreatic cancer. METHODS: Serum samples from five individuals with pancreatic cancer and five individuals without cancer were compared. Highly abundant serum proteins were depleted by

Yefei Rong; Dayong Jin; Chenrui Hou; Jianwen Hu; Wenchuan Wu; Xiaolin Ni; Dansong Wang; Wenhui Lou



Serum 25-hydroxyvitamin d levels and C-reactive protein in persons with human immunodeficiency virus infection.  


Human immunodeficiency virus (HIV) infection has frequently been associated with vitamin D deficiency as well as chronic inflammatory response. We tested the hypothesis of an independent relationship between serum concentrations of 25-hydroxyvitamin D [25(OH)D] and high-sensitivity C-reactive protein (CRP) in a cohort of HIV-positive people. A cross-sectional survey was conducted among 316 HIV-positive people (181 men and 135 women) aged 16 to 60 years residing in the Kathmandu Valley, Nepal. Serum high-sensitivity CRP concentrations and serum 25(OH)D levels were measured by the latex agglutination nephelometry method and the competitive protein-binding assay, respectively. The relationship between serum CRP concentrations and 25(OH)D serum level was assessed using multiple logistic regression analysis with adjustment of potential cardiovascular and HIV-related factors. The proportions of participants with 25(OH)D serum levels <20 ng/ml, 20-30 ng/ml, and ?30 ng/ml were 83.2%, 15.5%, and 1.3%, respectively. The mean 25(OH)D serum levels in men and women were 15.3 ng/ml and 14.4 ng/ml, respectively. Participants with a 25(OH)D serum level of <20 ng/ml had a 3.2-fold higher odds of high CRP (>3 mg/liter) compared to those with a 25(OH)D serum level of ?20 ng/ml (p=0.005). Men and women with a 25(OH)D serum level of <20 ng/ml had 3.2- and 2.7-fold higher odds of high CRP (>3 mg/liter), respectively, compared to those with a 25(OH)D serum level of ?20 ng/ml. The relationships remained significant only in men (p =0.02) but not in women (p=0.28). The risk of having a high level of inflammation (CRP>3 mg/liter) may be high among HIV-positive men and women with a 25(OH)D serum level of <20 ng/ml. PMID:23003113

Poudel-Tandukar, Kalpana; Poudel, Krishna C; Jimba, Masamine; Kobayashi, Jun; Johnson, C Anderson; Palmer, Paula H



Serum 25-Hydroxyvitamin D Levels and C-Reactive Protein in Persons with Human Immunodeficiency Virus Infection  

PubMed Central

Abstract Human immunodeficiency virus (HIV) infection has frequently been associated with vitamin D deficiency as well as chronic inflammatory response. We tested the hypothesis of an independent relationship between serum concentrations of 25-hydroxyvitamin D [25(OH)D] and high-sensitivity C-reactive protein (CRP) in a cohort of HIV-positive people. A cross-sectional survey was conducted among 316 HIV-positive people (181 men and 135 women) aged 16 to 60 years residing in the Kathmandu Valley, Nepal. Serum high-sensitivity CRP concentrations and serum 25(OH)D levels were measured by the latex agglutination nephelometry method and the competitive protein-binding assay, respectively. The relationship between serum CRP concentrations and 25(OH)D serum level was assessed using multiple logistic regression analysis with adjustment of potential cardiovascular and HIV-related factors. The proportions of participants with 25(OH)D serum levels <20?ng/ml, 20–30?ng/ml, and ?30?ng/ml were 83.2%, 15.5%, and 1.3%, respectively. The mean 25(OH)D serum levels in men and women were 15.3?ng/ml and 14.4?ng/ml, respectively. Participants with a 25(OH)D serum level of <20?ng/ml had a 3.2-fold higher odds of high CRP (>3?mg/liter) compared to those with a 25(OH)D serum level of ?20?ng/ml (p=0.005). Men and women with a 25(OH)D serum level of <20?ng/ml had 3.2- and 2.7-fold higher odds of high CRP (>3?mg/liter), respectively, compared to those with a 25(OH)D serum level of ?20?ng/ml. The relationships remained significant only in men (p?=0.02) but not in women (p=0.28). The risk of having a high level of inflammation (CRP>3?mg/liter) may be high among HIV-positive men and women with a 25(OH)D serum level of <20?ng/ml.

Poudel, Krishna C.; Jimba, Masamine; Kobayashi, Jun; Johnson, C. Anderson; Palmer, Paula H.



Seasonal influence on biochemical profile and serum protein electrophoresis for Boa constrictor amarali in captivity.  


Similarly to other reptiles, snakes are ectothermic animals and depend exclusively on the environment for the maintenance of their physiological, biochemical and immunological processes. Thus, changes in biochemical values can be expected due to seasonal influence. Twenty-two adult specimens of Boa constrictor amarali kept in captivity were used. Blood collections were done in two different seasons: winter (July 2004) and summer (January 2005) for the following assays: uric acid, aspartate aminotransferase (AST), glucose, cholesterol, total protein, and serum protein electrophoresis. The mean biochemical results found in summer and winter, respectively, were: 6.3 ± 3.4 and 11.3 ± 6.2 mg/dL for uric acid; 28.7 ± 12.4 and 20.7 ± 16.2 UI/L for AST; 26.3 ± 17 and 17.4 ± 6.8 mg/dL for glucose; 67.3 ± 30.2 and 69.7 ± 38.5 mg/dL for cholesterol; and 5.9 ± 1.6 and 5.9 ± 1.4 g/dL for total protein. Results regarding electrophoresis in summer and winter, respectively, were: 1.9 ± 0.7 and 2.4 ± 0.6 g/dL for albumin; 0.7 ± 0.2 and 0.5 ± 0.2 g/dL for ?-globulin; 1.5 ± 0.5 and 1.7 ± 0.6 g/dL for ?-globulin; and 1.8 ± 0.5 and 1.