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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



Protein electrophoresis - serum  


Total protein: 6.4 to 8.3 g/dL Albumin: 3.5 to 5.0 g/dL Alpha-1 globulin: 0.1 to 0.3 ... Consumptive coagulopathy Disseminated intravascular coagulation Increased gamma ... Acute infection Waldenstrom's macroglobulinemia ...


Immunochemical Study of Bovine Serum Proteins.  

National Technical Information Service (NTIS)

With the aid of immunological methods of analysis, this study characterizes certain protein fractions of the blood serum of cattle. Analyses indicate that serum proteins differ not only in their electrophoretic mobility, but also in their antigenic struct...

V. M. Kholod



Purine metabolites in serum of higher primates, including man  

Microsoft Academic Search

Using a new method for simultaneous quantification of hypoxanthine, xanthine, uric acid, and allantoin by means of high-pressure\\u000a liquid chromatography, purine metabolites of 18 species of higher primates, including man, have been determined. The data\\u000a thus produced indicate that the serum concentrations of purine metabolites in primates are influenced by nutrition, sexual\\u000a hormones, and the procedures used in catching the

G. Schreiber; W. Tiemeyer; Carola I. Flurer; H. Zucker I



Immunohistochemical demonstration of serum proteins in human cerebral gliomas  

Microsoft Academic Search

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

R. J. Seitz; W. Wechsler



Elemental Analysis of Human Serum and Serum Protein Fractions by Thermal Neutron Activation.  

National Technical Information Service (NTIS)

Some acplications of thermal neutron activation for the determination of elemental contents in human serum and human serum protein fractions are presented. First total serum is dealt with; second serum protein fractions obtained by gel filtration are desc...

J. R. W. Woittiez



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



Microchip-based capillary electrophoresis of human serum proteins  

Microsoft Academic Search

The separation and relative quantitation of human serum proteins is important to the clinical diagnosis of various states of disease. Microchip-based capillary electrophoresis (CE) of human serum proteins offers several advantages over sodium dodecyl sulfate-poly(acrylamide) gel electrophoresis and conventional CE methods, including decreased sample consumption and analysis time and the possibility of on-chip sample manipulation (dilution, labelling, etc.). The microchip

Christa L. Colyer; Shakuntala D. Mangru; D. Jed Harrison



Three automated quantitative assays for serum proteins.  


Three quantitative assays for detection of proteins are reported. One is an adaptation to the Auto-Analyzer of the widely used method of Lowry et al. In another procedure, the product of the reaction of proteins with a modified biuret reacts with the phosphomolybdic--phosphotungstic reagent of Folin and Ciocalteu. The third method, performed on hydrolyzed sample, is based on the measurement of proline (Pro) and the conversion of the Pro content into protein. The universal presence of proline in serum proteins suggested this assay. The values for total serum proteins as assayed by the three procedures are similar. The second and the third assay for proteins, described in this paper, can also be performed manually. Studies on interfering substances and their elimination as well as on the sensitivity of the assays are reported. PMID:7363455

Blumenkrantz, N



Fusion protein including of CD4  

US Patent & Trademark Office Database

Novel recombinant polypeptides are disclosed herein that include a CD4 polypeptide ligated at its C-terminus with a portion of an immunoglobulin comprising a hinge region and a constant domain of a mammalian immunoglobulin heavy chain. The portion or the IgG is fused at its C-terminus with a polypeptide comprising a tailpiece from the C-terminus of the heavy chain of an IgA antibody ara tailpiece from a C-terminus of the heavy chain of an IgM antibody. Also disclosed herein are methods for using these CD4 fusion proteins.



Serum protein electrophoresis in 147 dogs.  


Reference intervals for serum protein electrophoresis (SPE) were created from a group of 75 clinically healthy dogs and compared with SPE results obtained from clinical cases presented to the University of Bristol over an eight-and-a-half-year period. A total of 147 dogs, in which SPE had been performed, had complete case records available and thus met the inclusion criteria. Signalment and final diagnoses taken from the case records and SPE results were divided into normal and abnormal based on the newly established reference intervals. Cases were grouped according to the SPE protein fraction abnormalities and diagnosis using the DAMNITV classification system. Of the 147 cases, 140 (95.2 per cent) had abnormal SPE results. The most common protein fraction abnormality was decreased albumin (59.3 per cent) followed by a polyclonal increase in ? globulins (38.6 per cent). Decreased ?-1 globulins and increased ?-2 globulins were documented in 36.4 and 30.0 per cent of cases, respectively. The most common DAMNITV classification associated with abnormal SPE results was infectious/inflammatory disease, which was diagnosed in 79 of 140 cases (56.4 per cent). Monoclonal gammopathies were noted in eight dogs (5.7 per cent), and underlying lymphoproliferative disease was present in all cases where a diagnosis was achieved, including multiple myeloma (four dogs), splenic plasmacytoma (one dog), hepatic plasmacytoma (one dog) and lymphoma (one dog). PMID:21493443

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



Automated serum protein electrophoresis by Capillarys.  


Capillary zone electrophoresis (CZE) of serum proteins is increasingly gaining impact in clinical laboratories. In this report, we evaluate automated capillary zone electrophoresis by Capillarys (Sebia, France). Within-run and between-run imprecision for the five electrophoretic fractions was <2% and <6%, respectively. Data obtained with Capillarys correlated with results obtained with agarose gel electrophoresis and Paragon CZE 2000 (Beckman Coulter, USA). Analysis of serum obtained from patients with inflammation, nephrotic syndrome, bisalbuminemia, and alpha1-antitrypsin deficiency revealed that Capillarys was able to detect these abnormalities. Two hundred thirty eight samples were analyzed by agarose gel electrophoresis, Capillarys, capillary electrophoresis using Paragon CZE 2000 system, and immunofixation. Sample selection was based on the presence of a disturbed morphology (e.g., spike) of the protein profile or hypogammaglobulinemia on agarose gel electrophoresis and/or Capillarys. Immunofixation revealed the presence of a monoclonal protein, oligoclonal bands, polyclonal pattern, and a normal profile in, respectively, 89, 66, 19, and 64 samples. With Capillarys, Paragon, and agarose gel electrophoresis, a spike and/or disturbed morphology of the profile was found in 222, 182, and 180 samples, respectively. In these samples, immunofixation was negative in 73 (33%), 46 (25%), and 39 (22%) samples, respectively. These data indicate that Capillarys has a lower specificity than agarose gel electrophoresis and Paragon 2000. Of the 89 samples with a monoclonal protein, Capillarys, Paragon, and agarose gel electrophoresis failed to detect, respectively, three, three, and one monoclonal protein(s). Interferences by radio-opaque agents, complement degradation products, fibrinogen, and triglycerides are described. In conclusion, automated capillary zone electrophoresis with Capillarys provides for reproducible, rapid, and reliable serum electrophoresis. PMID:12812271

Bossuyt, Xavier; Lissoir, Bénédicte; Mariën, Godelieve; Maisin, Diane; Vunckx, Jozef; Blanckaert, Norbert; Wallemacq, Pierre



Electrohydrodynamic atomization of protein (bovine serum albumin).  


Bovine serum albumin (BSA) was chosen as a model protein. Three solutions of different concentrations of 5, 20 and 50 mg/ml were prepared, characterised and subjected to electrohydrodynamic atomization (EHDA). The 5 and 20 mg/ml solutions were atomized successfully and mode selection (M-S) maps were drawn for both concentrations to find out regions of stable cone-jet mode atomizaton. Droplet relics of these two solutions were investigated by electron microscopy. Samples were investigated by UV spectroscopy and circular dichroism (CD) spectroscopy before and after electrohydrodynamic atomization. We conclude that, particularly at the higher concentration of protein, EHDA does not result in significant structural change of BSA, and therefore is a processing route that can be considered for encapsulating drugs in proteins. PMID:16167100

Pareta, R; Brindley, A; Edirisinghe, M J; Jayasinghe, S N; Luklinska, Z B



Serum prolyl hydroxylase protein in experimental liver cancer.  


Total prolyl hydroxylase protein was measured in serum of rats fed with 0.06% of azodye, 3-methyl-4-dimethyl-aminoazobenzene (3-MeDAB). Serum enzyme protein values in the group progressing to primary hepatoma and those which had the tumors were significantly different (p less than 0.001) from the controls. The results suggest that the assay of serum prolyl hydroxylase protein may be an additional useful tool in the diagnosis of primary hepatic cancer. PMID:2839006

Bolarin, D M



Study of the effect of total serum protein and albumin concentrations on canine fructosamine concentration.  

PubMed Central

The relationship among serum fructosamine concentration and total serum protein and albumin concentrations were evaluated in healthy and sick dogs (diabetics and dogs with insulinoma were not included). Fructosamine was determined using a commercial colorimetric nitroblue tetrazolium method applied to the Technicon RA-500 (Bayer). Serum fructosamine concentration was not correlated to total protein in normoproteinemic (r = 0.03) and hyperproteinemic dogs (r = 0.29), but there was a high correlation (r = 0.73) in hypoproteinemic dogs. Similar comparison between serum fructosamine and albumin concentrations showed middle correlation (r = 0.49) in normoalbuminemic dogs and high degree of correlation (r = 0.67) in hypoalbuminemic dogs. These results showed the importance of recognizing serum glucose concentration as well as total serum protein and albumin concentrations in the assay of canine serum fructosamine concentration.

Loste, A; Marca, M C



Serum screening for oncogene proteins in workers exposed to PCBs  

Microsoft Academic Search

A cohort of 16 municipal workers engaged in cleaning oil from old transformers was examined for possible health effects from exposure to polychlorinated biphenyls (PCBs). In addition to the evaluation of routine clinical parameters (history, physical examination, liver function tests, serum triglycerides, serum PCB values), a new screening technique for the presence of oncogene proteins in serum using monoclonal antibodies

P W Brandt-Rauf; H L Niman



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.




Serum lipids and proteins in lactose malabsorption13  

Microsoft Academic Search

It has been suggested that dietary lactose may reduce the intestinal absorption of fat and protein in individuals with lactase deficiency. On the other hand, it is known that a high carbohydrate diet increases serum lipids. The purpose of this study was to examine whether there are differences iii the fasting serum lipid and protein concentrations between people with lactose

Timo Sahi; Jaakko Jussila; Seppo Sarna; Mauri Isokoski


Serum protein polymorphisms in four populations of Afghanistan.  

PubMed Central

Gene frequencies of the serum proteins third component of complement (C3) transferrin (Tf), haptoglobin (Hp), group specific component (Gc), serum cholinesterase (E1), alpha1-antitrypsin (Pi), beta2-glycoprotein I (Bg), and ceruloplasmin (Cp) in the Tajiks, Pushtoons, Hazaras, and Usbeks in Afghanistan were reported. Rare variants were observed in the C3, Tf, and Pi systems.

Rahimi, A G; Goedde, H W; Flatz, G; Kaifie, S; Benkmann, H G; Delbruck, H




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



Serum protein pattern in ewe with pregnancy toxemia  

Microsoft Academic Search

Pregnancy toxemia is a metabolic disease of pregnant ewes which causes significant economic losses in sheep industry. The\\u000a pathophysiology and metabolic changes of this disorder remain poorly understood. We conducted this study to describe the serum\\u000a protein pattern associated with the pregnancy toxemia in ewes. In this study, the electrophoretic pattern of serum proteins\\u000a of 15 ewes with naturally occuring

Gul Fatma Yarim; Gulay Ciftci



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.



Yield and Aging of Cheddar Cheeses Manufactured from Milks with Different Milk Serum Protein Contents  

Microsoft Academic Search

Whey proteins in general and specifically ?-lactoglo- bulin, ?-lactalbumin, and immunoglobulins have been thought to decrease proteolysis in cheeses manufac- turedfromconcentratedretentatesfromultrafiltration. The proteins found in whey are called whey proteins and are called milk serum proteins (SP) when they are in milk. The experiment included 3 treatments; low milk SP (0.18%), control (0.52%), and high milk SP (0.63%), and was

B. K. Nelson; D. M. Barbano



Grizzly bear corticosteroid binding globulin: Cloning and serum protein expression.  


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 of the stress status prior to capture in grizzly bears (Ursus arctos). To test this, a full-length CBG cDNA was cloned and sequenced from grizzly bear testis and polyclonal antibodies were generated for detection of this protein in bear sera. The deduced nucleotide and protein sequences were 1218 bp and 405 amino acids, respectively. Multiple sequence alignments showed that grizzly bear CBG (gbCBG) was 90% and 83% identical to the dog CBG nucleotide and amino acid sequences, respectively. The affinity purified rabbit gbCBG antiserum detected grizzly bear but not human CBG. There were no sex differences in serum total cortisol concentration, while CBG expression was significantly higher in adult females compared to males. Serum cortisol levels were significantly higher in bears captured by leg-hold snare compared to those captured by remote drug delivery from helicopter. However, serum CBG expression between these two groups did not differ significantly. Overall, serum CBG levels may be a better marker of chronic stress, especially because this protein is not modulated by the stress of capture and restraint in grizzly bears. PMID:20347821

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



Elevation of serum riboflavin carrier protein in hepatocellular carcinoma  

Microsoft Academic Search

Early detection of hepatocellular cancer (HCC), makes it surgically resectable with a potential for cure. The test most commonly used to detect HCC is the measurement of serum alpha-fetoprotein (AFP) levels. However, the AFP test is negative in HCC detection in more than 30% of the cases. Riboflavin carrier protein (RCP) is a growth and developmental protein, synthesized and secreted

Prakash N. Rao; Jeffrey Crippin; Edward Levine; Jay Hunt; Surendra Baliga; Luis Balart; Lowell Anthony; Madhuri Mulekar; Madhwa H. G. Raj



Analysis of glycans on serum proteins using antibody microarrays  

PubMed Central

Antibody arrays can be employed for the profiling glycan structures on proteins. Antibody arrays capture multiple, specific proteins directly from biological samples (such as serum), and lectin and glycan-binding antibodies probe the levels of specific glycans on the captured proteins. We use a practical method of partitioning microscope slides to enable the convenient processing of many detection reagents or samples. A critical first step in the procedure is the chemical derivatization of the glycans on the spotted capture antibodies, which prevents lectin binding to those glycans. We describe those methods along with the methods for preparing and treating serum samples, running the experiments, and designing and interpreting the experiments.

Chen, Songming; Haab, Brian B.



[Modification of placenta blood serum proteins under low temperature effect].  


Changes in environmental physical and chemical factors upon freeze-thawing and low temperature storage of biological samples can result in impairments of protein structures. This work specifies spontaneous and diamide-induced protein aggregations of placenta blood serum stored at -20 degrees and -196 degrees C during 2 years with SDS-PAGE. It was shown that storage of placenta blood serum at low temperatures did not cause any quantitative and qualitative changes in fraction distribution of proteins denatured with SDS in comparison to the native samples which were not frozen. Application of beta-mercaptoethanol revealed that placenta blood serum proteins upon freeze-thawing did not form spontaneous aggregates linked by disulphide bridges. Oxidation of amino acid sulfhydryl groups induced by diamide and accompanied by high molecular aggregate formation proved to be a quite effective way for indirect estimation of structural changes in protein upon low temperature effects. In samples thawed after low temperature storage the protein aggregation with 4 microM diamide was significantly higher than in native serum. These discrepancies between native and frozen-thawed samples are stipulated by impairments of protein structure under low temperature and increased in accessibility of reactive SH-groups of proteins for oxidation with diamide. Structural changes in placenta blood serum proteins, which caused by low temperatures and revealed by elevated sensibility to diamide-induced aggregate formation, did not depend on temperature (-20 degrees and -196 degrees C) and storage terms (2 years and 3 weeks). They reflect protein reaction to freeze-thawing processes and could be sequence of ice crystal formation which takes place in unprotected media. PMID:23789348

Fal'ko, O V; Zemlianskikh, N G; Lipina, O V; Prokopiuk, O S


Serum Resistin (FIZZ3) Protein Is Increased in Obese Humans  

Microsoft Academic Search

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




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



Electrophoretic and immunoelectrophoretic analysis of feline serum proteins.  

PubMed Central

Serum from 28 clinically healthy cats was subjected to agarose gel electrophoresis and the migration distance relative to albumin was determined. The reference values for the relative and absolute concentrations of each protein fraction were determined and compared to previous reports. The immunoelectrophoretic, crossed immunoelectrophoretic and crossed line immunoelectrophoretic pattern, of a pooled sample of serum from clinically normal cats was determined. The cross-reactivity between goat and/or rabbit monospecific antisera to human proteins and feline serum was determined using immunoelectrophoresis and crossed-line absorption immunoelectrophoresis. Feline alpha-2-macroglobulin, haptoglobin, B1C-globulin, IgG, albumin and ceruloplasmin cross reacted strongly with the monospecific antisera. Alpha-2-macroglobulin migrated anodal to haptoglobin. Lipoproteins and ceruloplasmin were studied using staining procedures described in man. Feline transferrin was precipitated with Rivanol. Images Fig. 4. Fig. 5.

Baker, R J; Valli, V E



Changes in serum protein profiles of chickens with tibial dyschondroplasia  

Technology Transfer Automated Retrieval System (TEKTRAN)

Differences in serum protein profiles were analyzed to identify biomarkers associated with a poultry leg problem named tibial dyschondroplasia (TD) that can cause lameness. We used a bead-based affinity matrix containing a combinatorial library of hexapeptides (ProteoMinerTM) to deplete high abundan...


Increased Serum Cholesteryl Ester Transfer Protein in Obese Children  

Microsoft Academic Search

Objective: To determine whether serum cholesteryl ester transfer protein (CETP), which is one of the physiologically active gene products secreted from adipose tissue, is increased and associated with atherogenic lipoprotein profile in obese children.Research Methods and Procedures: Subjects were 42 consecutive outpatient Japanese obese children, 29 boys and 13 girls, ranging in age from 5 to 14 years, and 25

Kohtaro Asayama; Hidemasa Hayashibe; Kazushige Dobashi; Norihiko Uchida; Takaya Nakane; Kohji Kodera; Akira Shirahata



Homology of beta Lactoglobulin, Serum Retinol-Binding Protein, and Protein HC  

Microsoft Academic Search

The milk protein beta -lactoglobulin has been extensively studied but its function has not been identified. A clue regarding the function of a protein can be obtained by discovering a genetic relationship with a protein of known function through comparisons of amino acid sequence. Such comparisons revealed that beta -lactoglobulin is similar to human serum retinol-binding protein and to another

Syed Pervaiz; Keith Brew




PubMed Central

Eight physicochemical factors which affect the uptake of lissamine green on filter paper impregnated with serum proteins have been examined, and their relevance to the staining of electrophoretically separated protein fractions is discussed. It is shown that grade of paper, weight of protein applied, separate and combined denaturation and staining time, temperature and concentration of staining solution, concentration of denaturant, and type of protein all influence the weight of dye absorbed per unit weight of applied protein, and must be rigidly standardized if valid quantitative results are to be obtained. Five sets of conditions are obtained for optimal staining and it is found that separation of denaturant from dye yields the best procedure. It is concluded that lissamine green is an excellent dye for the staining and quantitative estimation of separated protein fractions in paper electrophoresis, and that conditions can usually be arranged to produce a linear relation between dye uptake and protein concentration in an experimentally efficient manner.

Brackenridge, C. J.



Molecular forms of serum pancreatic stone protein in acute pancreatitis  

Microsoft Academic Search

Summary  \\u000a Conclusion: Elevation of serum pancreatic stone protein- (PSP) S1 suggests activation of trypsinogen in the pancreas. This information would prompt the start of intensive treatment and may\\u000a improve prognosis of acute pancreatitis (AP).\\u000a \\u000a \\u000a Background: PSP exists in two molecular forms, PSP-S2–5 and PSP-S1. PSP-S1 is produced by enzyme cleavage of PSP-S2–5 by trypsin. Total serum PSP rose in AP,

Yasuyuki Nakae; Satoru Naruse; Motoji Kitagawa; Hiroshi Ishiguro; Masanori Kato; Shinobu Hayakawa; Takaharu Kondo; Tetsuo Hayakawa



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



Pathology Case Study: Monoclonal Protein in Serum  

NSDL National Science Digital Library

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

Kelly, Robert; Torbenson, Michael



Standardisation of the quantitation of serum amyloid A protein (SAA) in human serum.  


An adequate method for standardising the quantitation of serum amyloid A protein (SAA) in human serum was developed. Acute phase high density lipoprotein3 (HDL3) was used as a standard. The concentration of the SAA in the standard was determined by the use of purified SAA. After protein determination, various concentrations of purified SAA were run on SDS-polyacrylamide gel together with the HDL3 standard containing an unknown amount of SAA amongst the apolipoproteins. From the standard curve obtained by pyridine extraction (Coomassie blue colour yield at A605 nm) the concentration of SAA in the HDL3 standard was determined. An established immunoradiometric assay (IRMA) for SAA was standardised with the HDL3. SAA concentrations in normal and acute phase sera were determined. PMID:4056404

Godenir, N L; Jeenah, M S; Coetzee, G A; Van der Westhuyzen, D R; Strachan, A F; De Beer, F C



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


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

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



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

PubMed Central

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

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



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



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

PubMed Central

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

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, Andre 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.



Expert systems for the interpretation of serum proteins.  


The ready availability of inexpensive desk-top computers with enormous disk storage has made the practicality of computer assisted medical interpretive software a reality. There seems little question that these programs could be of enormous help to physicians. However, there are daunting problems to their creation, including the lack of standards for clinical diagnostic precision or accuracy and paucity of helpful literature. As a result, the final products may be quite different. Little effort has been devoted in the laboratory to produce programs which could have great benefit in bridging the gap between laboratorians and clinicians. In a few circumscribed areas where the interpretation of laboratory measurements have been well studied in relation to patient demographics and to the final outcome, the impact has been enormous. The prime example is prenatal diagnosis of neural tube defects, and certain chromosomal and developmental abnormalities. Viewed as an obstacle by most people able and willing to attempt to create such programs is the omnipresence of necessary regulation. A brief overview of the general structure of a program to assist with the interpretation of serum proteins is given as a model in the perspective of current knowledge and state of the relevant literature. PMID:9877086

Ritchie, R F



[Effect of feed proteins on serum proteins in poultry infected with S. gallinarum pullorum].  


The effect was studied of the protein component of feed on the serum-protein profile in a Salmonella gallinarum pullorum infection on a model of birds of the Leghorn and Cornish breeds, participating in three age groups. In terms of feeding the birds were divided into two groups: I - controls, fed standard mixtures, and II - test birds, fed mixtures of low protein amounts. The birds were bled at definite intervals following their inoculations. The total proteins of the blood serum were determined, and the dynamics of the protein fractions of the serum was followed up through microelectrophoresis. Dispersion and statistical analysis was employed in data-processing. It was found that with the lowered amount of proteins in the ration of birds infected with Salmonella gallinarum pullorum there appeared changes in the protein spectrum - lower total protein, and a rise of the alfa - globulin fraction along with the depression and delayed synthesis of the beta- and gamma-globulin fractions of the blood serum. Greater were the changes in the serum-protein spectrum of birds at the age of 45 days, however, no breed differences were observed. PMID:7340110

Ganovska, M



Activation of Solubilized Steroid-Transforming Enzymes of Adrenal Microsomal Origin by Serum Proteins.  

National Technical Information Service (NTIS)

The experiments were designed to investigate the nature of the interaction between serum protein and the adrenal steroid 21-hydroxylase system, and to determine whether serum proteins also affected microsomal conversion of pregnenolone to progesterone whi...

M. A. Hamilton R. W. McCune S. Roberts



Characterization of Proteins in the Human Serum Proteome  

PubMed Central

This paper presents a multidimensional profile of the human serum proteome, produced by a two-dimensional protein fractionation system based on liquid chromatography followed by characterization with capillary electrophoresis (CE). The first-dimension separation was done by chromatofocusing over a pH range from 8.5 to 4.0, where proteins were separated by their isoelectric points (pI). In this dimension, fractions were collected based on pH. The first-dimension pI fractions were then resolved in the second dimension by high-resolution, reversed-phase chromatography with a gradient of trifluoroacetic acid (TFA) in acetonitrile and TFA in water. A selected protein fraction collected from the second dimension by time was characterized by CE for molecular-weight estimation and for presence of isoforms. Molecular-weight estimation was done by sodium dodecyl sulfate capillary gel electrophoresis, where proteins were separated in the range of 10,000–225,000 Da. Detection of isoforms was done by capillary isoelectric focusing over a pH range of 3–10. A selected second-dimension fraction that contained the putative serum iron–binding protein transferrin was analyzed by these two CE techniques for molecular-weight determination and the presence of isoforms. The combination of two-dimensional protein fractionation and CE characterization represents an advanced tool for proteomics.

Betgovargez, Edna; Knudson, Vita; Simonian, Michael H.



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

Microsoft Academic Search

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

B. C. Johnson; A. M. Swanson



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

Microsoft Academic Search

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

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



Association of increased serum heat shock protein 70 and C-reactive protein concentrations and decreased serum ? 2HS glycoprotein concentration with the syndrome of hemolysis, elevated liver enzymes, and low platelet count  

Microsoft Academic Search

The primary aim of this study was to determine serum Hsp70 concentrations in HELLP syndrome. We measured also the serum concentrations of three acute phase proteins: C-reactive protein (CRP), ?2-macroglobulin (AMG) and ?2-HS glycoprotein (AHSG). Ten severe preeclamptic patients with HELLP syndrome, 20 severe preeclamptic patients without HELLP syndrome and 20 normotensive, healthy pregnant women were included in this case-control

Attila Molvarec; Zoltán Prohászka; Bálint Nagy; László Kalabay; János Szalay; Georg Füst; István Karádi; János Rigó



Identification of M protein from filter paper using serum protein and immunofixation electrophoresis.  


Serum protein electrophoresis (SPE) and immunofixation electrophoresis (IFE) are standard methods for detection and monitoring of monoclonal (M) proteins. However, these tests are rarely available in the remote areas, especially in developing countries. Transportation of fresh serum (FS) samples is also usually inconvenient. This study investigated M-protein identification using serum blot on filter paper (FP). SPE and IFE were performed on FS and FP specimens using the Sebia Hydrasys automated electrophoresis system. Statistical analyses were conducted to assess sample stability and agreement of FS vs FP. The FP method showed good agreement with the FS method. The r values for correlation of albumin levels-?(1), ?(2), ?, and ? (%)-between FP and FS samples in SPE were all more than 0.95 (P < .01). IFE displayed no significant difference between those 2 methods in the identification of M protein. The FP method demonstrated an accurate and reproducible alternative to FS for identification of M protein using SPE and IFE. PMID:23010716

Wu, Yonghua; Yang, Xu; Wang, Tiancheng; Wang, Haining; Li, Zhenrong



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



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



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.



Acute-phase proteins and serum immunoglobulins in ankylosing spondylitis.  

PubMed Central

The erythrocyte sedimentation rate (ESR) and the serum acute-phase proteins (APP), C-reactive protein (CRP), fibrinogen, 9th component of complement (C9), and alpha, antitrypsin were measured on 231 occasions in 80 patients with ankylosing spondylitis and compared with those in 30 controls. APP levels did not correlate with clinical assessment of disease activity. However, there were significant correlations between CRP, C9, and fibrinogen (p = less than 0.01), suggesting that these APP may be more reliable indicators of disease activity. The mean values of the APP in those patients with a peripheral arthritis were significantly higher than in those with pelvospondylitis alone for ESR (p less than 0.01), CRP (p less than 0.01), and fibrinogen (p less than 0.05). The only significant difference between those patients with an iritis and those with only pelvospondylitis was an elevated CRP in the iritis group (p less than 0.01). This suggests that a peripheral arthritis is the most important cause of an elevated ESR or APP in ankylosing spondylitis. Serum immunoglobulins were also measured and they showed a significant elevation of IgA in all 3 patients groups, there being no difference between each group. Serum IgG was raised only in those patients with an iritis or peripheral arthritis, the IgM levels being within the normal range for all patient groups.

Laurent, M R; Panayi, G S



Serum Resistance in Haemophilus ducreyi Requires Outer Membrane Protein DsrA  

PubMed Central

Haemophilus ducreyi is resistant to killing by normal serum antibody and complement. We discovered an H. ducreyi outer membrane protein required for expression of serum resistance and termed it DsrA (for “ducreyi serum resistance A”). The dsrA locus was cloned, sequenced, and mutagenized. An isogenic mutant (FX517) of parent strain 35000 was constructed and characterized, and it was found to no longer express dsrA. FX517 was at least 10-fold more serum susceptible than 35000. DsrA was expressed by all strains of H. ducreyi tested, except three naturally occurring, avirulent, serum-sensitive strains. FX517 and the three naturally occurring dsrA-nonexpressing strains were complemented in trans with a plasmid expressing dsrA. All four strains were converted to a serum-resistant phenotype, including two that contained truncated lipooligosaccharide (LOS). Therefore, serum resistance in H. ducreyi does not require expression of full-length LOS but does require expression of dsrA. The dsrA locus from eight additional H. ducreyi strains was sequenced, and the deduced amino acid sequences were more than 85% identical. The major difference between the DsrA proteins was due to the presence of one, two, or three copies of the heptameric amino acid repeat NTHNINK. These repeats account for the variability in apparent molecular mass of the monomeric form of DsrA (28 to 35 kDa) observed in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Since DsrA is present in virulent strains, is highly conserved, and is required for serum resistance, we speculate that it may be a virulence factor and a potential vaccine candidate.

Elkins, Christopher; Morrow, K. John; Olsen, Bonnie



Serum Protein Electrophoresis in the Evaluation of Lytic Bone Lesions  

PubMed Central

Serum protein electrophoresis (SPEP) is often obtained at the initial evaluation of a radiolucent bone lesion of unknown etiology. The results are considered convincing evidence of the presence or absence of a plasma cell neoplasm. The sensitivity and specificity of the SPEP have not been reported in this clinical scenario. Our purpose is to assess the diagnostic value of the SPEP in the initial work-up of the radiolucent bone lesion. We identified 182 patients undergoing evaluation of a radiolucent bone lesion that included tissue biopsy and an SPEP value. We then calculated the sen-sitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of SPEP as a diagnostic test for a plasma cell neo-plasm in this clinical scenario. Forty-six of 182 (25.3%) patients in our series were diagnosed with a plasma cell neo-plasm by histopathologic analysis. The sensitivity of SPEP was 71% and the specificity was 83%. PPV was 47% and NPV was 94%. When analyzing only those presenting with multiple lesions, the percentage of patients diag-nosed with multiple myeloma increased to 44.7% (34 of 76 patients). The SPEP, however, did not have a substantially increased diagnostic accuracy with sensitivity of 71%, specificity 79%, PPV 40% and NPV 93%. SPEP lacks sensitivity and positive predictive value to provide a definitive diagnosis of myeloma in radiolucent bone lesions, but has a high negative predictive value which may make it useful in ruling out the disease. We recommend that this test either be performed in conjunction with urine electrophoresis, immunofixation electro-phoresis and free light chain assay, or after biopsy confirming the diagnosis of myeloma.

Nystrom, Lukas M.; Buckwalter, Joseph A.; Syrbu, Sergei; Miller, Benjamin J.



Serum protein electrophoresis in the evaluation of lytic bone lesions.  


Serum protein electrophoresis (SPEP) is often obtained at the initial evaluation of a radiolucent bone lesion of unknown etiology. The results are considered convincing evidence of the presence or absence of a plasma cell neoplasm. The sensitivity and specificity of the SPEP have not been reported in this clinical scenario. Our purpose is to assess the diagnostic value of the SPEP in the initial work-up of the radiolucent bone lesion. We identified 182 patients undergoing evaluation of a radiolucent bone lesion that included tissue biopsy and an SPEP value. We then calculated the sen-sitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of SPEP as a diagnostic test for a plasma cell neo-plasm in this clinical scenario. Forty-six of 182 (25.3%) patients in our series were diagnosed with a plasma cell neo-plasm by histopathologic analysis. The sensitivity of SPEP was 71% and the specificity was 83%. PPV was 47% and NPV was 94%. When analyzing only those presenting with multiple lesions, the percentage of patients diag-nosed with multiple myeloma increased to 44.7% (34 of 76 patients). The SPEP, however, did not have a substantially increased diagnostic accuracy with sensitivity of 71%, specificity 79%, PPV 40% and NPV 93%. SPEP lacks sensitivity and positive predictive value to provide a definitive diagnosis of myeloma in radiolucent bone lesions, but has a high negative predictive value which may make it useful in ruling out the disease. We recommend that this test either be performed in conjunction with urine electrophoresis, immunofixation electro-phoresis and free light chain assay, or after biopsy confirming the diagnosis of myeloma. PMID:24027470

Nystrom, Lukas M; Buckwalter, Joseph A; Syrbu, Sergei; Miller, Benjamin J



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.




Microsoft Academic Search

This study was conducted for qualitative analysis of serum proteins separated by SDS-PAGE and stained by Coomassie Brilliant Blue R-250 in order to describe the preliminary identification of serum proteins that may act as diagnostic marker in benign prostatic hyperplasia. Serum samples of 23 biopsy confirmed cases of benign prostatic hyperplasia and normal controls of similar age group were subjected

Saima Naz; Sarah Ahmad; Farkhanda Ghafoor



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



Improved medium and culture conditions for clonal growth with minimal serum protein and for enhanced serum-free survival of Swiss 3T3 cells  

Microsoft Academic Search

Summary  Improved culture conditions have been developed that will support clonal growth of Swiss mouse embryo 3T3 cells at concentrations\\u000a of serum protein as low as 125?g\\/ml. Survival of the cells under completely protein-free conditions also is enhanced greatly.\\u000a The improvements that made these results possible include: (a) use of medium MCDB 402, which was developed specifically for\\u000a Swiss 3T3 cells

Gary D. Shipley; Richard G. Ham



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



Increased serum level of Nup88 protein is associated with the development of colorectal cancer.  


Nucleoporin88 (Nup88) has been shown to be overexpressed in a wide variety of malignancies including colorectal cancer (CRC). However, no study about serum Nup88 in human CRC was reported. Therefore, in this study, we investigated the level of serum Nup88 protein and its relationships with clinicopathological variables in CRC. The serum concentration of Nup88 protein was determined by a quantitative sandwich enzyme-linked immunosorbent assay (ELISA) in 118 pre-operative serum samples, 66 post-operative and 96 healthy controls. Among the patients, the levels of CEA (n = 91) and CA19-9 (n = 87) in the pre-operative serum were measured, and DNA sequencing was performed in 12 CRCs and 2 samples from non-cancerous colon tissue. In the same patients, the level of pre-operative serum Nup88 was significantly higher than that of post-operative Nup88 (P = 0.021). Furthermore, the level of pre-operative Nup88 was positively related to the depth of tumor invasion (P = 0.002) and advanced stage (P = 0.001). The level of pre-operative Nup88 in the left colon tended to be higher than that in the right colon and the rectum (P = 0.063). DNA sequencing results showed that there were two single nucleotide polymorphisms, distributed in exon 6 (NM_002532.3:c.1044G>A (ACG-ACA, Thr ? Thr) and exon 10 (NM_002532.3:c.1389A>T, CCA-CCT, Pro ? Pro). Serum Nup88 might be a candidate for a new biomarker implicated in the development and aggressiveness of CRCs. PMID:21863385

Zhao, Zeng-Ren; Zhang, Li-Jing; Wang, Yuan-Yuan; Li, Fang; Wang, Ming-Wei; Sun, Xiao-Feng



Purification of lipopolysaccharide-binding protein from bovine serum.  


Lipopolysaccharide-binding protein (LBP) plays a central role in presentation of bacterial-derived lipopolysaccharide (LPS; endotoxin) to leukocytes such as macrophages and neutrophils. Interaction of LBP with LPS is significant because LBP-LPS complexes promote activation of leukocytes and the immune system, which results in enhanced secretion of a spectrum of proinflammatory cytokines. An improved, simplified method was used to purify bovine LBP from serum. Methodology consisted of ion-exchange chromatography using Bio-Rex 70 resin, followed by gel-filtration chromatography (Sephacryl S-200 resin) of a selected ion-exchange fraction (0.22-0.50 M NaCl). Densitometric scans on silver-stained polyacrylamide gels of chromatographically-derived proteins indicated up to 88.7% purity of the resultant 64kD protein (bovine LBP) in the cleanest fractions. The isoelectric point of bovine LBP was determined to be 6.8. Identity of the protein was substantiated by western-blot analysis, and by N-terminus amino acid sequence analysis with favorable comparison to published sequence data from rabbit, human, and murine LBP Identity was corroborated by use of purified bovine LBP in bioassays which demonstrated enhanced tissue factor expression of LPS (1 ng ml(-1)-stimulated bovine alveolar macrophages. Tissue factor expression was inhibitable in these assays using anti-CD14 monoclonal antibodies, which is also consistent with LBP-mediated activation of cells. When bovine LBP was heated at 56 degrees C for 30 min, the biological activity was reduced by 50% in the macrophage-based bioassays. Biological activity of bovine LBP was completely destroyed by heating at 62 degrees C for 30 min, which compared favorably with data resulting from use of fetal bovine serum. PMID:8792567

Bochsler, P N; Yang, Z; Murphy, C L; Carroll, R C



Serum acute phase proteins in dogs with symptomatic esophageal spirocercosis.  


Canine spirocercosis (CS) is a helminthic infection caused by the nematode Spirocerca lupi. The clinical hallmark of the disease is esophageal dysphagia due to parasite-induced esophageal nodules. Currently, there is limited information on the involvement of serum acute phase proteins (APPs) in the symptomatic CS. The objective of this study was to investigate whether C-reactive protein (CRP), serum amyloid A (SAA), haptoglobin (Hp) and albumin are involved in CS, and if their concentrations measured on admission reflect the severity of benign esophageal lesions. Nineteen dogs with spontaneous symptomatic esophageal spirocercosis and 7 clinically healthy dogs were studied retrospectively. The most consistently increased APP in the symptomatic dogs was Hp (95% of the dogs), followed by CRP (68%). The SAA concentrations were infrequently increased (5% of the dogs), while albumin concentrations were decreased in 58% of the affected dogs. The dogs with spirocercosis had significantly higher median concentrations of Hp (p=0.0001) and CRP (p=0.02) compared to healthy dogs. Median albumin concentrations did not differ between the two groups of dogs. The median concentrations of Hp, CRP and albumin did not differ significantly between the dogs having a single or multiple esophageal nodules. The results of this study indicate that in symptomatic CS, Hp and CRP are significantly and consistently increased, while SAA and albumin may be of limited value as diagnostic markers. No association was established between the concentrations of Hp, CRP and albumin measured on admission and the number of esophageal nodules. PMID:22683300

Mylonakis, Mathios E; Ceron, Jose J; Leontides, Leonidas; Rallis, Tim S; Koutinas, Alexander F



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



Association of urinary polycyclic aromatic hydrocarbons and serum C-reactive protein  

Microsoft Academic Search

The association of 9 urinary monohydroxy polycyclic aromatic hydrocarbons (OH-PAHs) with serum C-reactive protein (CRP) was investigated using the National Health and Nutrition Examination Survey (NHANES) 2003–2004. The unweighted number of participants included was 999, which represented 139,362,776 persons in the non-institutionalized US population. In adjusted logistic regressions, two OH-PAHs, 2-hydroxyphenanthrene and 9-hydroxyfluorene, were associated with elevated CRP (>3mg\\/l). Logistic

Charles J. Everett; Dana E. King; Marty S. Player; Eric M. Matheson; Robert E. Post; Arch G. Mainous III



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

PubMed Central

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

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



Identification of Protein Carbonyls in serum of the fetal and neonatal pig.  

Technology Transfer Automated Retrieval System (TEKTRAN)

Oxidation of serum proteins leads to non-reversible carbonyl formation which alters their function and has long been associated with stress-related disease processes. The primary objective of this study was to identify oxidized serum protein biomarkers of metabolic stress in baby pigs. Protein carb...


Enrichment of low molecular weight serum proteins using acetonitrile precipitation for mass spectrometry based proteomic analysis.  


A rapid acetonitrile (ACN)-based extraction method has been developed that reproducibly depletes high abundance and high molecular weight proteins from serum prior to mass spectrometric analysis. A nanoflow liquid chromatography/tandem mass spectrometry (nano-LC/MS/MS) multiple reaction monitoring (MRM) method for 57 high to medium abundance serum proteins was used to characterise the ACN-depleted fraction after tryptic digestion. Of the 57 targeted proteins 29 were detected and albumin, the most abundant protein in serum and plasma, was identified as the 20th most abundant protein in the extract. The combination of ACN depletion and one-dimensional nano-LC/MS/MS enabled the detection of the low abundance serum protein, insulin-like growth factor-I (IGF-I), which has a serum concentration in the region of 100 ng/mL. One-dimensional sodium dodecyl sulfate/polyacrylamide gel electrophoresis (SDS-PAGE) analysis of the depleted serum showed no bands corresponding to proteins of molecular mass over 75 kDa after extraction, demonstrating the efficiency of the method for the depletion of high molecular weight proteins. Total protein analysis of the ACN extracts showed that approximately 99.6% of all protein is removed from the serum. The ACN-depletion strategy offers a viable alternative to the immunochemistry-based protein-depletion techniques commonly used for removing high abundance proteins from serum prior to MS-based proteomic analyses. PMID:18803344

Kay, Richard; Barton, Chris; Ratcliffe, Lucy; Matharoo-Ball, Balwir; Brown, Pamela; Roberts, Jane; Teale, Phil; Creaser, Colin



Comparison of composition and sensory properties of 80% whey protein and milk serum protein concentrates.  


Milk serum protein concentrates (SPC) are proteins found in cheese whey that are removed directly from milk. Because SPC are not exposed to the cheese-making process, enzymatic or chemical reactions that can lead to off-flavors are reduced. The objectives of this study were to identify and compare the composition, flavor, and volatile components of 80% protein SPC and whey protein concentrates (WPC). Each pair of 80% SPC and WPC was manufactured from the same lot of milk and this was replicated 3 times. At each replication, spray-dried product from each protein source was collected. Commercial 80% WPC were also collected from several manufacturers for sensory and volatile analyses. A trained sensory panel documented the sensory profiles of the rehydrated powders. Volatile components were extracted by solid-phase microextraction and solvent extraction followed by solvent-assisted flavor evaporation with gas chromatography-mass spectrometry and gas chromatography-olfactometry. Consumer acceptance testing of acidified 6% protein beverages made with 80% SPC and WPC produced in the pilot plant and with WPC from commercial sources was conducted. The SPC was lower in fat and had a higher pH than the WPC produced in the pilot plant or commercial WPC. Few sensory differences were found between the rehydrated SPC and WPC manufactured in this study, but their flavor profiles were distinct from the flavor of rehydrated commercial WPC. The pilot-plant WPC had higher concentrations of lipid oxidation products compared with SPC, which may be related to the higher fat content of WPC. There was a large difference in appearance between 80% SPC and WPC: solutions of SPC were clear and those of WPC were opaque. Concentrations of lipid oxidation products in commercial WPC were generally higher than those in pilot-plant SPC or WPC. Sensory profiles of the peach-flavored protein beverage included cereal, free fatty acid, and soapy flavors and bitter taste in beverages made from pilot-plant products, whereas cardboard flavors were detected in those made with commercial WPC. Consumer liking scores for the beverages made with SPC were ranked highest or equally high with beverages made with WPC for aroma, appearance, and mouthfeel, but the beverages made with SPC had lower flavor and overall liking scores compared with beverages made with 3 of the 4 WPC. PMID:20412896

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



Proteomic analysis of maternal serum in down syndrome: identification of novel protein biomarkers.  


Down syndrome (DS) is the most prevalent chromosomal disorder, accounting for significant morbidity and mortality. Definitive diagnosis requires invasive amniocentesis, and current maternal serum-based testing requires a false-positive rate of about 5% to detect 85% of affected pregnancies. We have performed a comprehensive proteomic analysis to identify potential serum biomarkers to detect DS. First- and second-trimester maternal serum samples of DS and gestational age-matched controls were analyzed using multiple, complementary proteomic approaches, including fluorescence 2-dimensional gel electrophoresis (2D-DIGE), 2-dimensional liquid chromatography-chromatofocusing (2D-CF), multidimensional protein identification technology (MudPIT; LC/LC-MS/MS), and MALDI-TOF-MS peptide profiling. In total, 28 and 26 proteins were differentially present in first- and second-trimester samples, respectively. Of these, 19 were specific for the first trimester and 16 for the second trimester, and 10 were differentially present in both trimesters. Analysis of MALDI-TOF-MS peptide profiles with pattern-recognition software also discriminated between DS and controls in both trimesters, with an average recognition capability approaching 96%. A majority of the biomarkers identified are serum glycoproteins that may play a role in cellular differentiation and growth of fetus. Further characterization and quantification of these markers in a larger cohort of subjects may provide the basis for new tests for improved DS screening. PMID:17373838

Nagalla, Srinivasa R; Canick, Jacob A; Jacob, Thomas; Schneider, Kimberly A; Reddy, Ashok P; Thomas, Archana; Dasari, Surendra; Lu, Xinfang; Lapidus, Jodi A; Lambert-Messerlian, Geralyn M; Gravett, Michael G; Roberts, Charles T; Luthy, David; Malone, Fergal D; D'Alton, Mary E



Baseline Staging of Melanoma with Unknown Primary Site: The Value of Serum S100 Protein and Positron Emission Tomography  

Microsoft Academic Search

Background: Baseline staging is important in all melanoma types, including melanoma with unknown primary site (MUP). Staging includes different examination strategies, each with different accuracy. Objective: To determine the value of serum S100 protein levels and positron emission tomography (PET) in the baseline staging of MUP. Methods: Twenty patients with MUP were evaluable for the analysis between 1996 and 2007

Patrick A. Oberholzer; Mirjana Urosevic; Hans C. Steinert; Reinhard Dummer



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



Discovery of serum biomarkers of alcoholic fatty liver in a rodent model: C-reactive protein  

PubMed Central

Background Excessive consumption of alcohol contributes to alcoholic liver disease. Fatty liver is the early stage of alcohol-related liver disease. The aim of this study was to search for specific serological biomarkers of alcoholic fatty liver (AFL) compared to healthy controls, non-alcoholic fatty liver (NAFL) and liver fibrosis in a rodent model. Methods Serum samples derived from animals with AFL, NAFL, or liver fibrosis were characterized and compared using two-dimensional differential gel electrophoresis. A matrix-assisted laser desorption ionization-time of flight tandem mass spectrometer in conjunction with mascot software was used for protein identification. Subsequently, Western blotting and flexible multi-analyte profiling were used to measure the expressions of the putative biomarkers present in the serum of animals and clinical patients. Results Eight differential putative biomarkers were identified, and the two most differentiated proteins, including upregulated C-reactive protein (CRP) and downregulated haptoglobin (Hp), were further investigated. Western blotting validated that CRP was dramatically higher in the serum of AFL compared to healthy controls and other animals with liver disease of NAFL or liver fibrosis (p < 0.05). Moreover, we found that CRP and Hp were both lower in liver fibrosis of TAA-induced rats and clinical hepatitis C virus-infected patients. Conclusion The results suggest that increased levels of CRP are an early sign of AFL in rats. The abnormally elevated CRP induced by ethanol can be used as a biomarker to distinguish AFL from normal or otherwise diseased livers.



HPLC-DAD protein kinase inhibitor analysis in human serum.  


We here describe an HPLC-DAD method to analyse different protein kinase inhibitors. Potential applications of this method are pharmacokinetic studies and therapeutic drug monitoring. Optimised chromatography conditions resulted in a very good separation of seven inhibitors (vatalanib, bosutinib, canertinib, tandutinib, pazopanib, dasatinib - internal standard and erlotinib). The good sensitivity makes this method competitive with LC/MS/MS. The separation was performed with a Lichrospher 100-5 RP8, 250 mm × 4 mm column maintained at 30 ± 1 °C, and with a mobile phase of 0.05 M H(3)PO(4)/KH(2)PO(4) (pH=2.3)-acetonitrile (7:3, v/v) at a flow rate of 0.7 mL/min. A simple and fast sample preparation sequence with liquid-liquid extraction led to good recoveries (73-90%) of all analytes. The recovery hardly reached 50% only for pazopanib. This method can also be used for targeted protein kinase inhibitor quantification. A perfect linearity in the validated range (20-10,000 ng/mL) and an LOQ of 20 ng/mL were achieved. The relative standard deviations and accuracies of all examined drug concentrations gave values much lower than 15% both for between- and within-batch calculations. All analysed PKIs were stable for 6 months in a 1mg/mL dimethyl sulfoxide stock solution. Vatalanib, bosutinib and erlotinib were also stable in human serum in the whole examined concentration range. PMID:22425385

Dziadosz, Marek; Lessig, Rüdiger; Bartels, Heidemarie




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



Insulin or sulfonylurea treatments of the diabetics differentially affect erythrocyte membrane and serum enzymes and extent of protein glycosylation.  


Erythrocyte membrane protein glycosylation increase by 3.4 fold in diabetes. Insulin or sulfonylurea treatment did not reduce the extent of glycosylation. The serum protein glycosylation was comparable in all the groups including control. Erythrocyte membrane Na(+),K(+)-ATPase activity decreased in the diabetics; only insulin treatment partly restored the activity. Erythrocyte membrane acetylcholinesterase activity decreased only in the sulfonylurea treated group. Serum butyrylcholinesterase activity was relatively low in the diabetic and insulin treated diabetic groups. The Km and Vmax of the two components of Na(+),K(+)-ATPase from erythrocyte membranes were differently affected in the diabetic and the two treatment groups. The Vmax of acetylcholinesterase decreased only in the sulfonylurea treated group. Diabetic states resulted in decreased Vmax of components I and II of serum butyrylcholinesterase. In insulin-treated diabetics, component II was absent. Sulfonylurea group resembled diabetics.In vitro incubation with insulin differentially affected the Na(+),K(+)-ATPase and serum butyrylcholinesterase activities. PMID:23105297

Dave, K R; Patel, T H; Katyare, S S



Biomarkers of Eosinophil Involvement in Allergic and Eosinophilic Diseases: review of phenotypic and serum markers including a novel assay to quantify levels of soluble Siglec-8  

PubMed Central

There remains considerable controversy in the management of eosinophilic disorders, mainly due to a paucity of information regarding the clinical interpretation of total blood eosinophil counts versus surface activation markers versus eosinophil-derived or eosinophil-influencing mediator levels. Regrettably, few tests have been validated that define a unique clinical or prognostic phenotype that is more useful than simply monitoring total blood eosinophil counts. In this manuscript, phenotypic (cell surface) markers, along with serum and tissue-based markers that have been examined in the context of disease activity, are reviewed. We also report the development of a novel assay for detecting soluble Siglec-8 (sSiglec-8), a protein likely derived largely from eosinophils, as a potential serum biomarker. The assay consists of a competitive ELISA using a recombinant Siglec-8-Fc fusion protein. The goal of this preliminary study was to determine if sSiglec-8 is a useful biomarker that differentiates among patients with various eosinophil-associated diseases. In the final analysis, it is fair to say that further research is sorely needed to fully understand and validate the utility of various biomarkers, including sSiglec-8, before their use in clinical practice can be recommended with confidence.

Na, Ho Jeong; Hamilton, Robert G.; Klion, Amy D.



Irreversible Effects of Serum Proteins on Beta-Lactam Antibiotics  

PubMed Central

The chromogenic cephalosporin nitrocefin (87/312) demonstrates rapid and visible instability to serum from many species. This phenomenon was distinct from serum binding, being significantly slower. Destruction of another cephalosporin, 10485, by serum appeared to account for some anomalous results during investigation into its human pharmacokinetics. Many cephalosporins of very different structures also showed serum instability, unrelated to their degrees of serum binding as measured by plate assay. Extrapolation could not be made from one species to another with regard to either binding or instability. Small changes in the chemical structures of the 3- and 7-substituents of the cephalosporins made profound changes in their susceptibility to serum attack. The decomposition is pH dependent, occurring more slowly at acid pH, and is due to a high-molecular-weight component of serum that resists boiling for several minutes. Isoelectric focusing of serum from several animal species gave various species-specific bands that decomposed nitrocefin. The inactivation of nitrocefin was not entirely parallel with that of 10485 and was inhibited by it. All other ?-lactam compounds tested also inhibited the reaction, much greater concentrations usually being necessary when the inhibitor was stable to serum. The complex that causes breakdown of the ?-lactam compounds is not necessarily the same as the one causing serum binding. It is postulated that serum may affect most other ?-lactam antibiotics in a similar way, although in most cases, this only occurs to a very slight extent.

O'Callaghan, Cynthia H.



Serum Protein MALDI Profiling to Distinguish Upper Aerodigestive Tract Cancer Patients From Control Subjects  

Microsoft Academic Search

Background: There are no reliable blood markers for the early detection and monitoring of aerodigestive tract tumors. Recent studies have suggested that serum protein patterns may be able to distinguish cancer patients from control subjects. Methods: We used matrix-assisted laser desorption and ionization (MALDI) mass spectroscopy to obtain serum protein patterns from patients with head and neck cancer (n 99)

David Sidransky; Rafael Irizarry; Joseph A. Califano; Xianbin Li; Hening Ren; Nicole Benoit; Li Mao



A comparison of depletion versus equalization for reducing high-abundance proteins in human serum.  


In this work three methods to diminish the content of most highly abundant proteins in human serum have been studied and compared. Protein depletion with ACN or DTT and protein equalization with the ProteoMiner(™) (PM) have been assessed by 1-D gel electrophoresis and MS. After treatment 5, 18 and 9 major proteins within the 20 most abundant proteins in serum were identified for the ACN, DTT and PM methods, respectively. The ACN method was efficient for depleting high molecular weight proteins, over 75?KDa, resulting in 10±4% (n=3) of the total protein content remaining in the depleted serum. In addition, 75% of the proteins belonging to the group of the 20 most abundant proteins were not detected, making this depletion strategy a cheap alternative to expensive commercial tools regularly used for removing high abundance proteins from serum. The ACN extract was found rich in apolipoproteins. The dithithreitol method promotes the precipitation of proteins rich in disulfide bonds, mainly albumin, with 73±7% (n=3) of the total protein content remaining in the depleted serum, which was found rich in immunoglobulins. The PM method compresses the dynamic range of the serum proteins, rendering an extract containing 16±2% (n=3) of the total initial protein content. The extract was found to be rich in both apolipoproteins and immunoglobulins. As a general rule the DTT and PM methods provide a compression of the dynamic range of serum protein concentrations while the ACN method allows an effective depletion of the protein fraction above 72?KDa. PMID:21997478

Fernández, Carolina; Santos, Hugo M; Ruíz-Romero, Cristina; Blanco, Francisco J; Capelo-Martínez, José-Luis



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

PubMed Central

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 CTD-matched women without silicone implants, were assessed in the primary study; other groups were used for additional comparisons. Routine serum protein determinations and high-sensitivity protein electrophoresis and immunofixation electrophoresis were performed for detection of paraproteins. Women with silicone implants, either with or without CTD, had significantly lower serum total protein and ?1-globulin, ?2-globulin, ?-globulin, ?-globulin, and IgG levels compared with those without silicone implants. There was no significant difference, however, in the frequency of paraproteinemia between women with silicone implants and CTD (9.5%) and age-matched and CTD-matched women without silicone implants (5.4%) (odds ratio, 1.82; 95% confidence interval, 0.51–6.45). Paraprotein isotypes were similar in the two groups, and the clinical characteristics of the 13 women with paraproteinemia were comparable with an independent population of 10 women with silicone breast implants, CTD, and previously diagnosed monoclonal gammopathies. In summary, this first comprehensive study of serum proteins in women with silicone implants and CTD found no substantially increased risk of monoclonal gammopathy. Women with silicone implants, however, had unexpectedly low serum globulin and immunoglobulin levels, with or without the subsequent development of CTD. The causes and clinical implications of these findings require further investigation.

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



Decrease in Serum Protein Carbonyl Groups Concentration and Maintained Hyperhomocysteinemia in Patients Undergoing Bariatric Surgery  

Microsoft Academic Search

Background  Human obesity is associated with oxidative stress but the factors contributing to the increase of reactive oxygen species\\u000a (ROS) production remain unknown. We evaluated the association between serum homocysteine concentration, which may increase\\u000a ROS production, and serum protein carbonyl groups concentration before and after bariatric surgery.\\u000a \\u000a \\u000a \\u000a Methods  Serum protein carbonyl groups and serum homocysteine concentrations, as well as obesity markers, were

T. Sledzinski; E. Goyke; R. T. Smolenski; Z. Sledzinski; J. Swierczynski



Proteomic Analysis of Nitrated and 4Hydroxy2- Nonenal-Modified Serum Proteins During Aging  

Microsoft Academic Search

Using proteomic techniques, we investigated peroxynitrite (ONOO? ) and 4-hydroxy-2-nonenal (HNE) modified serum proteins from young and old Fischer 344 rats. Two-dimensional gel electrophoresis\\/western blot analysis of nitrotyrosine and HNE-histidine revealed that serum proteins were differentially modified by ONOOand HNE. Among them, 16 of the modified proteins, identified by matrix-assisted laser desorption\\/ionization-time of flight mass spectrom- etry (MALDI-TOF MS), are

Chul Hong Kim; Yani Zou; Dae Hyun Kim; Nam Deuk Kim; Byung Pal Yu; Hae Young Chung



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



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



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


Oxidation of serum proteins can lead to carbonyl formation that alters their function and is often associated with stress-related diseases. As it is recommended that all pigs reared in modern production facilities be given supplemental iron at birth to prevent anemia, and metals can catalyze the carbonylation of proteins, the primary objective of this study was to determine whether standard iron dextran treatment was associated with enhanced serum protein oxidation in newborn piglets. Piglets were treated with 100 mg of iron dextran intramuscularly either on the day of birth, or on the third day after birth. Blood samples were collected from piglets 48 or 96 h after treatment and serum was harvested. For quantification, serum protein carbonyls were converted to hydrazones with dinitrophenyl hydrazine and analyzed spectrophotometrically. To identify and determine relative distribution of carbonylated proteins, serum protein carbonyls were derivatized with biotin hydrazide, separated by two-dimensional polyacrylamide gel electrophoresis, stained with avidin-fluorescein and identified by mass spectrometry. The standard iron dextran treatment was associated with no increase in total oxidized proteins if given either on the first or third day of life. In addition, with a few noted exceptions, the overall distribution and identification of oxidized proteins were similar between control and iron dextran-treated pigs. These results indicate that while iron dextran treatment is associated with a marked increase in circulating iron, it does not appear to specifically induce the oxidation of serum proteins. PMID:22436157

Caperna, T J; Shannon, A E; Blomberg, L A; Garrett, W M; Ramsay, T G



Effects of Adherence, Activation and Distinct Serum Proteins on the In Vitro Human Monocyte Maturation Process  

Microsoft Academic Search

Elutriator-punfied human monocytes were cultured in a serum-free (SF) medium, and various serum proteins and functional activating agents were assessed for their effects on the in vitro maturation of human monocytes to macrophages. Following 3 days of suspension culture in Teflon Iabware, 60% of the monocytes were easily recovered. When varying concentrations of human AB serum (HuAB) were employed, human

Yukio Akiyama; Richard Griffith; Paul Miller; G. W. Stevenson; Stacy Lund; D. J. Kanapa; Henry C. Stevenson


Serum proteins and immunoglobulins in oral submucous fibrosis  

Microsoft Academic Search

Patients suffering from Oral Submucous Fibrosis (OSMF) have significantly elevated levels of serum globulins and immunoglobulin\\u000a IgG. (P.< 0.001). Hyperglobulinaemia favours the possibility of OSMF being an autoimmune disorder.

A. G. Phatak



Human Serum Haeme-albumin: An Allosteric ‘Chronosteric’ Protein  

Microsoft Academic Search

Serum albumin (SA) participates in plasma haeme scavenging. Sequestering the haeme, SA accounts for most of the antioxidant\\u000a capacity of plasma. In turn, serum haeme albumin (SA-haeme) displays ligand-binding and pseudoenzymatic properties. Recently,\\u000a engineered SA-haeme has been proposed as an O2 carrier not only for red blood cell substitutes but also as an O2-therapeutic agent. Eventually, SA-haeme could be considered

Mauro Fasano; Gabriella Fanali; Riccardo Fesce; Paolo Ascenzi


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

Microsoft Academic Search

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

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



Serum Levels of the Adipokine Adipocyte Fatty Acid–binding Protein Are Increased in Preeclampsia  

Microsoft Academic Search

BackgroundPreeclampsia (PE) is a serious complication of pregnancy which is associated with an increased future metabolic and cardiovascular risk for both mother and newborn. Recently, adipocyte fatty acid–binding protein (AFABP) was introduced as a novel adipokine, serum levels of which independently correlate with the development of the metabolic syndrome and cardiovascular disease in humans. In this study, we investigated serum

Mathias Fasshauer; Jeannette Seeger; Theresa Waldeyer; Susanne Schrey; Thomas Ebert; Jürgen Kratzsch; Ulrike Lössner; Matthias Blüher; Michael Stumvoll; Renaldo Faber; Holger Stepan



Association of Serum C-Reactive Protein (CRP) with Some Nutritional Parameters of Maintenance Hemodialysis Patients  

Microsoft Academic Search

Malnutrition and inflammation are common in hemodialysis patients, and are usually closely associated. Serum C-reactive protein (CRP) concentrations have been found to be significantly elevated in hemodialysis patients and reflects chronic inflammation, and as an acute-phase reactant, is a sensitive and independent marker of malnutrition. To investigate the association of serum CRP level with some nutritional variables in diabetic and



Serum phospholipid transfer protein activity and genetic variation of the PLTP gene  

Microsoft Academic Search

The inverse relationship between serum levels of high density lipoproteins (HDL) and risk of coronary heart disease is well established. The phospholipid transfer protein (PLTP) promotes the transfer of phospholipids between lipoproteins and modulates HDL size and composition. It thus plays a central role in HDL metabolism. Serum PLTP activity was measured in 400 healthy Finnish individuals in order to

Esa Tahvanainen; Matti Jauhiainen; Harald Funke; Erkki Vartiainen; Jouko Sundvall; Christian Ehnholm



Transient increase of serum Clara cell protein (CC16) after exposure to smoke  

Microsoft Academic Search

OBJECTIVES: Smoke inhalation is a well known cause of airways injury in firefighting personnel. The aim of this study was to evaluate whether toxic effects of smoke on the respiratory tract can be detected by measuring Clara cell protein (CC16), a recently described serum marker of lung function. METHODS: CC16 was measured by a sensitive latex immunoassay in the serum

A Bernard; C Hermans; G Van Houte



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  

NASA Astrophysics Data System (ADS)

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

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



Serum protein binding of diazepam in maternal and foetal serum during pregnancy.  

PubMed Central

1 The serum binding capacity for diazepam was significantly lower in pregnancy and there was a linear correlation with gestational age. 2 The binding of diazepam was not correlated to albumin during pregnancy. 3 In cord sera there was a significantly reduced binding capacity for diazepam with albumin levels of less than 40 g/l.

Lee, J N; Chen, S S; Richens, A; Menabawey, M; Chard, T



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



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

PubMed Central

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

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



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

Microsoft Academic Search

BACKGROUND: The aim of the study was to optimize protocols for finding and identifying serum proteins that are differentially expressed in persons with normal glucose tolerance (NGT) compared to individuals with type 2 diabetes mellitus (T2DM). Serum from persons with NGT and persons with T2DM was profiled using ProteinChip arrays and time-of-flight mass spectra were generated by surface enhanced laser

Tea Sundsten; Michael Eberhardson; Michael Göransson; Peter Bergsten



An electrophoretic taxonomic study on serum proteins of Acanthobrama marmid, Leuciscus cephalus, and Chondrostoma regium  

Microsoft Academic Search

In this study, native polyacrylamide gel electrophoresis (Native-PAGE) and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) were applied to the serum proteins of Leuciscus cephalus, Acanthobrama marmid and Chondrostoma regium (Cyprinidae) fish taken from) the Karakaya Dam (Malatya, Turkey). The electrophoregrams showed that there were similarities and differences in the molecular weight (MW) of the serum proteins among the three species.

Muhittin Yilmaz; H. Ramazan Yilmaz; Ali Alas


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



Peptide sequences that target proteins to lysosomes for enhanced degradation during serum withdrawal  

SciTech Connect

Ribonuclease A (RNase A) microinjected into human diploid fibroblasts is degraded with a half-life of 80-100 h in the presence of serum, but its half-life declines to 40 h when cells are deprived of serum. This increased degradation in response to serum withdrawal results from an increased rate of uptake of microinjected RNase A from the cytosol into lysosomes. The cells' ability to recognize RNase A for this enhanced degradation is based on some feature of amino acids 1-20. Covalent linkage of S-peptide to other proteins results in enhanced lysosomal degradation of the conjugate in response to serum withdrawal. The authors have further defined the essential region of RNase S-peptide to be within residues 7-11, KFERQ. Coinjection of radiolabeled RNase A and excess unlabeled pentapeptide specifically blocks the enhanced degradation of RNase A. Affinity-purified polyclonal IgGs raised against KFERQ immunoprecipitate 25-30% of radiolabeled cytosolic proteins from human fibroblasts. Pulse-chase experiments indicate that these proteins are preferentially degraded upon serum withdrawal while degradation of nonimmuno-precipitated proteins is unaffected. These results suggest that peptide sequences similar to KFERQ are contained within cellular proteins and that such sequences target proteins to lysosomes for enhanced degradation during serum withdrawal.

Dice, J.F.; Backer, J.M.; Chiang, H.L.



Serum C-reactive protein and leptin for assessment of nutritional status in patients on maintenance hemodialysis.  


Nutrition is one of the key parameters in predicting morbidity and mortality in patients with end-stage renal disease (ESRD) on hemodialysis. Body weight, body mass index, and visceral protein levels (serum protein, albumin, prealbumin, and transferrin) have traditionally been used as markers for nutritional status. Serum leptin and C-reactive protein (CRP), have been recently added to the list of markers for nutritional status. This study was a comparative assessment of serum leptin and CRP for nutritional status in patients with ESRD on maintenance hemodialysis. A total of 40 patients with ESRD on maintenance hemodialysis and a similar number of age-, gender-, and BMI-matched healthy individuals were studies. Complete medical history was obtained and relevant clinical examination including anthropometry was carried out. All the individuals were subjected to routine investigations and special investigations (serum leptin and CRP). Data were analyzed using Student's t-test and correlation was found using Pearson's correlation coefficient. Mean value of serum leptin for the study group (1.44 ± 0.72 ng/ml) was found to be significantly higher than that of the control group (0.68 ± 0.55 ng/ml). In addition, we also observed a positive correlation between serum leptin and BMI (r = 0.350, P<0.05). For CRP, we observed that the study group (3.93 ± 1.20 mg/ml) had a significantly higher value vis-ŕ-vis the control group (0.28 ± 0.24 mg/ml). However, CRP and BMI did not show a significant correlation. Based on the above observations, we conclude that serum leptin is a better biomarker than CRP for assessing nutritional status in patients with ESRD on maintenance hemodialysis. PMID:23440668

Kaur, S; Singh, N P; Jain, A K; Thakur, A



Measurement of serum proteins during attacks of ulcerative colitis as a guide to patient management.  

PubMed Central

Serial measurements of 11 serum proteins have been made throughout 39 admissions of 36 patients to hospital for the treatment of acute attacks of ulcerative colitis. There was a striking correlation between rapid changes in C-reactive protein and pre-albumin concentrations and the clinical response to medical treatment. Measurements of the alpha1-acid glycoprotein, albumin, and total serum protein concentrations at the time of admission were found to correlate with the outcome of the attack. Measurement of these proteins provides a useful guide to the management of patients with attacks of ulcerative colitis.

Buckell, N A; Lennard-Jones, J E; Hernandez, M A; Kohn, J; Riches, P G; Wadsworth, J



Serum protein fractionation using supported molecular matrix electrophoresis.  


Supported molecular matrix electrophoresis (SMME), in which a hydrophilic polymer such as PVA serves as a support within a porous PVDF membrane, was recently developed. This method is similar to cellulose acetate membrane electrophoresis but differs in the compatibility to glycan analysis of the separated bands. In this report, we describe the first instance of the application of SMME to human serum fractionation, and demonstrate the differences with serum fractionation by cellulose acetate membrane electrophoresis. The SMME membrane exhibited almost no EOF during electrophoresis, unlike the cellulose acetate membrane, but afforded comparative results for serum fractionation. The visualization of each fraction was achieved by conventional staining with dye such as Direct Blue-71, and objective quantification was obtained by densitometry after inducing membrane transparency with 1-nonene. Immunostaining was also achieved. Moreover, mass spectrometric analysis of both N-linked and O-linked glycans from the separated bands was demonstrated. Serum fractionation and glycan profiling of each fraction using SMME will enable novel insights into the relationships between various glycosylation profiles and disease states. PMID:23765906

Dong, Weijie; Matsuno, Yu-Ki; Kameyama, Akihiko



Analyses of intricate kinetics of the serum proteome during and after colon surgery by protein expression time series  

Microsoft Academic Search

Monitoring changes in serum protein expression in response to acute events such as trauma, infection or drug intervention may reveal key proteins of great value in predicting recovery or treatment response. Concerted actions of many proteins are expected. Proteins sharing similar expression changes may function in the same physiological process. As a model we analyzed expression changes in serum of

Han Roelofsen; Gloria Alvarez-Llamas; Martijn Dijkstra; Rainer Breitling; Klaas Havenga; Johan Bijzet; Wouter Zandbergen; Marcel P. de Vries; Rutger J. Ploeg; Roel J. Vonk



Preparation of Solid-Phase Immunosorbents by Coupling Human Serum Proteins to Cyanogen Bromide-Activated Agarose  

PubMed Central

The method of preparing solid-phase immunosorbents by covalently attaching proteins from whole human serum to cyanogen bromide-activated agarose has been investigated to determine optimum concentrations of cyanogen bromide and protein, and the optimum pH for the maximum attachment of proteins from serum. Two systems in which the above immunosorbents have proved useful are described: the removal of antibodies to normal serum proteins from anti-hepatitis B serum and the removal of light chain antibodies from anti-human immunoglobulin M serum. Images

Sipe, J. D.; Schaefer, F. V.



Interaction of liposomal formulations of meta-tetra(hydroxyphenyl)chlorin (temoporfin) with serum proteins: protein binding and liposome destruction.  


mTHPC is a non polar photosensitizer used in photodynamic therapy. To improve its solubility and pharmacokinetic properties, liposomes were proposed as drug carriers. Binding of liposomal mTHPC to serum proteins and stability of drug carriers in serum are of major importance for PDT efficacy; however, neither was reported before. We studied drug binding to human serum proteins using size-exclusion chromatography. Liposomes destruction in human serum was measured by nanoparticle tracking analysis (NTA). Inclusion of mTHPC into conventional (Foslip(®)) and PEGylated (Fospeg(®)) liposomes does not affect equilibrium serum protein binding compared with solvent-based mTHPC. At short incubation times the redistribution of mTHPC from Foslip(®) and Fospeg(®) proceeds by both drug release and liposomes destruction. At longer incubation times, the drug redistributes only by release. The release of mTHPC from PEGylated vesicles is delayed compared with conventional liposomes, alongside with greatly decreased liposomes destruction. Thus, for long-circulation times the pharmacokinetic behavior of Fospeg(®) could be influenced by a combination of protein- and liposome-bound drug. The study highlights the modes of interaction of photosensitizer-loaded nanovesicles in serum to predict optimal drug delivery and behavior in vivo in preclinical models, as well as the novel application of NTA to assess the destruction of liposomes. PMID:22607362

Reshetov, Vadzim; Zorin, Vladimir; Siupa, Agnieszka; D'Hallewin, Marie-Ange; Guillemin, François; Bezdetnaya, Lina



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

PubMed Central

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

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



[Spectrofluorometric analysis of protein fractions of blood serum of health donors and patients with kidney diseases].  


A comparison was made of ultra-violet fluorescence characteristics of albumin enriched protein fractions of serum blood of 35 glomerulonephritis (GN) patients, 30 uremic haemodialysis patients, and 40 donors. It has been found that fluorescence spectra of the albumin enriched fractions of serum from GN patients are blue shifted as compared with those of donors' serum preparations. At the same time fluorescence spectra of the albumin enriched fractions of donors' serum are similar, while those of GN patients vary significantly. The ratio of fluorescence intensities at 320 and 365 nm (A = I320/I365), characterizing the spectrum position is 1.27 +/- 0.05, for protein preparations of donors, while that for GN patients varies within the limits of 1.3-2.1. Fluorescence spectra of protein fractions from blood serum of uremic patients are red shifted (A = 0.77-1.29) in comparison with those of donors' blood. Chromatographic investigations show that protein preparations of GN patients' blood contain monomers and dimers of albumin, that can be divided according to their molecular masses or hydrophobic properties of the surface. Joint chromatographic and spectral analysis allowed to distinguish up to six albumin enriched fractions. A protein fraction with blue fluorescence spectrum was obtained by gel-filtration and ion exchange chromatography. A higher concentration of this fraction determines a high value of parameter A, that is typical for protein preparations of GN patients' blood. Amino acid sequence shows that one component of this fraction is beta-haptoglobin. PMID:9821250

Parkhomenko, T V; Klytsenko, O A; Shavlovski?, M M; Poletaev, A I; Kuznetsova, I M; Turoverov, K K



Significance of serum protein S100 levels in screening for melanoma metastasis: does protein S100 enable early detection of melanoma recurrence?  


A number of recent reports suggest serum protein S100 as a prognostic parameter in patients with metastatic melanoma. In the present study, serum protein S100 was investigated as a tumour marker for screening for melanoma metastasis in patients attending regular follow-up examinations. During the period from September 1997 to December 1998, serum protein S100 levels were measured by an immunoluminometric assay in 411 consecutive high risk melanoma patients (666 samples) and in 120 control subjects. Melanoma patients with resected primary tumours with a tumour thickness of 1.5 mm or more with resected metastasis were included in the study. Overall, 41 of the 411 patients developed metastasis during the period of observation. According to the distribution of protein S100 levels, the following different cut-off values were examined: 0.08 microg/l (95 percentile of the control group) and 0.13 microg/l (95 percentile of the group of melanoma patients without metastasis). The test efficiency for protein S100 as a diagnostic test for the detection of metastasis was highest for the cut-off value of 0.13 microg/l. In eight of the 41 patients (19.5%), elevation of protein S100 was the first sign of recurrence. Of the 41 patients with metastatic disease, 13 had elevated protein S100, giving a sensitivity of 0.32. The specificity for the detection of metastasis was 0.96. In eight of the 14 patients (57%) who developed distant metastasis, elevated S100 values were the first sign of tumour progression. In conclusion, determination of serum protein S100 levels enables earlier detection of distant metastasis in patients at high risk for metastasis. The impact on survival time needs to be investigated in follow-up studies. PMID:11095406

Schlagenhauff, B; Schittek, B; Ellwanger, U; Stroebel, W; Blum, A; Schwarz, M; Rassner, G; Garbe, C



A Serum Protein Profile Predictive of the Resistance to Neoadjuvant Chemotherapy in Advanced Breast Cancers*  

PubMed Central

Prediction of the responses to neoadjuvant chemotherapy (NACT) can improve the treatment of patients with advanced breast cancer. Genes and proteins predictive of chemoresistance have been extensively studied in breast cancer tissues. However, noninvasive serum biomarkers capable of such prediction have been rarely exploited. Here, we performed profiling of N-glycosylated proteins in serum from fifteen advanced breast cancer patients (ten patients sensitive to and five patients resistant to NACT) to discover serum biomarkers of chemoresistance using a label-free liquid chromatography-tandem MS method. By performing a series of statistical analyses of the proteomic data, we selected thirteen biomarker candidates and tested their differential serum levels by Western blotting in 13 independent samples (eight patients sensitive to and five patients resistant to NACT). Among the candidates, we then selected the final set of six potential serum biomarkers (AHSG, APOB, C3, C9, CP, and ORM1) whose differential expression was confirmed in the independent samples. Finally, we demonstrated that a multivariate classification model using the six proteins could predict responses to NACT and further predict relapse-free survival of patients. In summary, global N-glycoproteome profile in serum revealed a protein pattern predictive of the responses to NACT, which can be further validated in large clinical studies.

Hyung, Seok-Won; Lee, Min Young; Yu, Jong-Han; Shin, Byunghee; Jung, Hee-Jung; Park, Jong-Moon; Han, Wonshik; Lee, Kyung-Min; Moon, Hyeong-Gon; Zhang, Hui; Aebersold, Ruedi; Hwang, Daehee; Lee, Sang-Won; Yu, Myeong-Hee; Noh, Dong-Young



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.



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

Microsoft Academic Search

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

Christopher M. R. Bax; David L. Bloxam



Search for protein markers for serum diagnostics of tumors by analysis of microRNA expression profiles  

Microsoft Academic Search

New algorithm of bioinformatic search for potential serum tumor markers has been worked out, including: (1) identification\\u000a of microRNAs with the level of synthesis most evidently and often decreasing in tumors; (2) search for target mRNAs regulated\\u000a by microRNAs; (3) selection of targets encoding secretory proteins; (4) comparative analysis of transcription level of the\\u000a targets in normal and tumor tissues.

Yu. A. Bukurova; I. G. Nikitina; S. L. Khankin; G. S. Krasnov; N. A. Lisitsyn; V. L. Karpov; S. F. Beresten



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

PubMed Central

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

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



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

PubMed Central

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

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



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



The Promoter Polymorphism in the Eosinophil Cationic Protein Gene and Its Influence on the Serum Eosinophil Cationic Protein Level  

Microsoft Academic Search

Asthma is characterized by reversible airway obstruction and airway ECP is a member of the eosinophil-associated RNase fam- inflammation. Serum levels of eosinophil cationic protein (ECP) ily, and the gene is located on human chromosome 14q11.2. might reflect eosinophilic airway inflammation and asthma activity. The eosinophil peroxidase and eosinophil-derived neuro- However, serum ECP levels are not elevated in some patients

Emiko Noguchi; Asushi Iwama; Kazunori Takeda; Tetsuya Takeda; Masashi Kamioka; Kunio Ichikawa; Toshiko Akiba; Tadao Arinami; Masanao Shibasaki


Increased serum levels of non-collagenous matrix proteins (cartilage oligomeric matrix protein and melanoma inhibitory activity) in marathon runners  

Microsoft Academic Search

Objective Marathon runners have an increased risk of developing joint disease. During and after a 42-km run, elevation of multiple cytokines occurs in the blood, reflecting inflammatory processes. We compared this cytokine response with serum levels of cartilage oligomeric matrix protein (COMP) and melanoma inhibitory activity (MIA), two markers for joint metabolism and\\/or damage.Methods Serum from eight endurance-trained runners was

M. Neidhart; U. Müller-Ladner; W. Frey; A. K. Bosserhoff; P. C. Colombani; P. Frey-Rindova; K. M. Hummel; R. E. Gay; H.-J. Häuselmann; S. Gay



Influence of colostral quality on serum proteins in dairy calves raised in smallholder farms in Thailand.  


The objective of this study was to analyze the influence of colostral quality on serum proteins in calves. Samples were collected from visited farms in Kasetsart University Veterinary Teaching Hospital at Kamphaeng Saen and Nong Pho Animal Hospital. In total, 35 dairy farms contributed 80 dams and calves' samples. Colostrum samples from 80 dairy cows and blood samples from their calves were taken to evaluate colostral immunoglobulins (Ig) and immunoglobulin G (IgG), and calf serum protein and IgG. Total colostral Ig, colostral and serum IgG, and serum protein were measured by a colostrometer, single radial immunodiffusion, and refractrometer, respectively. Immunoglobulin G and serum protein concentrations increased in the 1st day after birth, and maximum concentrations were seen in the 2nd day and then decreased in the 7th and 14th days. Average?±?SD total colostral IgG concentrations at calving date and at 1 and 2 days after calving were 93.85?±?33.89, 37.11?±?23.51, and 17.23?±?9.4 mg/mL, respectively. The profile of total Ig and IgG concentrations in colostrum had a similar pattern, with the maximum concentrations obtained in calving date and rapidly decreased thereafter. Low IgG concentrations were seen in the 7th and 14th day after calving. The calves that were fed with high quality colostrum had higher serum protein at 1 day of age, 7.49?±?1.01 g/dL, than calves fed with low quality colostrum, 6.40?±?0.86 g/dL (P?serum protein after first colostrum feeding of high and low quality colostrum was 1.55?±?1.07 and 0.81?±?0.69 g/dL, respectively (P?=?0.02). PMID:23645514

Kananub, Suppada; Rukkwamsuk, Theera; Arunvipas, Pipat



Yield and aging of Cheddar cheeses manufactured from milks with different milk serum protein contents.  


Whey proteins in general and specifically beta-lactoglobulin, alpha-lactalbumin, and immunoglobulins have been thought to decrease proteolysis in cheeses manufactured from concentrated retentates from ultrafiltration. The proteins found in whey are called whey proteins and are called milk serum proteins (SP) when they are in milk. The experiment included 3 treatments; low milk SP (0.18%), control (0.52%), and high milk SP (0.63%), and was replicated 3 times. The standardized milk for cheese making of the low milk SP treatment contained more casein as a percentage of true protein and more calcium as a percentage of crude protein, whereas the nonprotein nitrogen and total calcium content was not different from the control and high SP treatments. The nonprotein nitrogen and total calcium content of the milks did not differ because of the process used to remove the milk SP from skim milk. The low milk SP milk contained less free fatty acids (FFA) than the control and high milk SP treatment; however, no differences in FFA content of the cheeses was detected. Approximately 40 to 45% of the FFA found in the milk before cheese making was lost into the whey during cheese making. Decreasing the milk SP content of milk by 65% and increasing the content by 21% did not significantly influence general Cheddar cheese composition. Higher fat recovery and cheese yield were detected in the low milk SP treatment cheeses. There was more proteolysis in the low milk SP cheese and this may be due to the lower concentration of undenatured beta-lactoglobulin, alpha-lactalbumin, and other high molecular weight SP retained in the cheeses made from milk with low milk SP content. PMID:16291609

Nelson, B K; Barbano, D M



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.



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

Microsoft Academic Search

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

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



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



Mutations and polymorphisms of the gene of the major human blood protein, serum albumin.  


We have tabulated the 77 currently known mutations of the familiar human blood protein, serum albumin (ALB). A total of 65 mutations result in bisalbuminemia. Physiological and structural effects of these mutations are included where observed. Most of the changes are benign. The majority of them were detected upon clinical electrophoretic studies, as a result of a point mutation of a charged amino acid residue. Three were discovered by their strong binding of thyroxine or triiodothyronine. A total of 12 of the tabulated mutations result in analbuminemia, defined as a serum albumin concentration of <1 g/L. These were generally detected upon finding a low albumin concentration in patients with mild edema, and involve either splicing errors negating translation or premature stop codons producing truncated albumin molecules. A total of nine mutations, five of those with analbuminemia and four resulting in variants modified near the C-terminal end, cause frameshifts. Allotypes from three of the point mutations become N-glycosylated and one C-terminal frameshift mutation shows O-glycosylation. PMID:18459107

Minchiotti, Lorenzo; Galliano, Monica; Kragh-Hansen, Ulrich; Peters, Theodore



Gold-peptide nanoconjugate cellular uptake is modulated by serum proteins.  


Gold nanoparticles (Au NPs, 20 nm) were conjugated with two different cysteine-terminated peptides. Radio-ligand binding studies were conducted to characterize Au NP-peptide binding, suggesting both covalent and noncovalent interactions. The interactions of serum proteins with Au NP-peptide nanoconjugates were determined using gel electrophoresis and dynamic light scattering. Serum proteins rapidly bound the nanoconjugates (15 minutes). The cellular uptake of free peptides and nanoconjugates into mouse myogenic (Sol8) cells was investigated in the absence or presence of serum. In the absence of serum, peptides presented as nanoconjugates showed significantly higher intracellular fluorescence signals compared to those in the presence of serum (P < 0.05), suggesting that serum proteins inhibit Au NP-mediated peptide delivery. The cellular uptake of nanoconjugates was also confirmed using transmission electron microscopy. These data suggest that Au NP-peptide nanoconjugates are a useful platform for intracellular delivery of therapeutics. However, a deeper understanding of the mechanisms regulating their uptake and intracellular trafficking is needed. PMID:22051699

Wang, Guankui; Papasani, Madhusudhan R; Cheguru, Pallavi; Hrdlicka, Patrick J; Hill, Rodney A



Serum bone gla protein (BGP) and other markers of bone mineral metabolism in postmenopausal osteoporosis  

Microsoft Academic Search

Summary  Bone gla protein, the vitamin K-dependent protein synthesized by osteoblasts and measured in blood by radioimmunoassay, has\\u000a been used as an index of the rate of bone turnover. The relationship of bone gla protein with other markers of bone mineral\\u000a metabolism was determined in 31 untreated postmenopausal women with the osteoporotic syndrome. In addition to serum osteocalcin\\u000a (BGP) we measured

F. Ismail; S. Epstein; R. Pacifici; D. Droke; S. B. Thomas; L. V. Avioli



Intravenous streptokinase treatment and serum C-reactive protein in patients with acute myocardial infarction  

Microsoft Academic Search

Serum concentrations of C-reactive protein were studied in 23 patients with acute myocardial infarction. In 14 patients who did not receive thrombolytic treatment there was a linear relation between infarct size (determined by serial creatine kinase-MB determinations and thallium-201 isotope emission tomography) and the C-reactive protein response. The correlation coefficient between the concentration-time integrals of creatine kinase-MB and C-reactive protein

K Pietilä; A Harmoinen; L Pöyhönen; M Koskinen; J Heikkila; R Ruosteenoja



Pulsed Electric Field Induced Aggregation of Food Proteins: Ovalbumin and Bovine Serum Albumin  

Microsoft Academic Search

Ovalbumin (OVA), bovine serum albumin (BSA), and a mixture of the two proteins (OVA?+?BSA) in solution were exposed to pulsed\\u000a electric field (PEF) to investigate the protein interaction and aggregation. The results demonstrated the self-aggregation\\u000a of OVA through disulfide bond due to the exposure of sulfhydryl groups and intermolecular disulfide interactions when PEF\\u000a intensity exceeded 25 kV cm?1. However, no protein self-aggregation

Wei Zhao; Ruijin Yang


Erythrocyte Sedimentation Rate, C-Reactive Protein Level, and Serum Amyloid A Protein for Patient Selection and Monitoring of AntiTumor Necrosis Factor Treatment in Ankylosing Spondylitis  

Microsoft Academic Search

Objective. To study the usefulness of erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), and serum amyloid A (SAA) for response prediction and monitoring of anti-tumor necrosis factor (anti-TNF) treatment in ankylosing spondylitis (AS) patients. Methods. Patients were included consecutively before starting etanercept or infliximab treatment. ASsessment in Ankylosing Spondylitis (ASAS) response, defined as a 50% improvement or an absolute improvement

Vries de M. K; Eijk van I. C; I. E. Bruinsma; Mike J. L. Peters; Michael T. Nurmohamed; Ben A. C. Dijkmans; Bouke P. C. Hazenberg; Gerrit J. Wolbink



Effective stabilization of curcumin by association to plasma proteins: human serum albumin and fibrinogen.  


The use of curcumin as an effective wound healing agent is of significant interest currently. It is well established that curcumin undergoes rapid degradation in physiological buffer by hydrolysis. The means by which curcumin is stabilized at the wound site to enable healing is poorly understood because blood plasma is composed of approximately 92% water. Plasma proteins, which constitute the remaining 6-8%, has been shown to stabilize curcumin. It is, however, still unclear which proteins are responsible for this phenomenon. In this study, the effects of major plasma proteins, which include human serum albumin (HSA), fibrinogen, immunoglobulin G (IgG), and transferrin, on stabilizing curcumin are investigated. In particular, we investigate their effects on the hydrolysis of curcumin at pH 7.4. In the presence of both transferrin and IgG, curcumin continues to undergo rapid hydrolysis but this reaction is suppressed by the presence of either HSA or fibrinogen with an impressive yield of approximately 95%. Furthermore, the binding constants of curcumin to HSA and fibrinogen are on the order of 10(4) and 10(5) M(-1), respectively. The binding constants of transferrin and IgG, however, are at least 1 order of magnitude less than those of HSA and fibrinogen. The results support that strong binding occurs at the hydrophobic moieties of HSA and fibrinogen, excluding water access. Therefore, strong interactions with HSA and fibrinogen inhibit hydrolysis of curcumin and in turn lead to effective suppression of degradation. PMID:19320475

Leung, Mandy H M; Kee, Tak W



Homology Modeling and Domain Interactions in Fetal Serum Protein  

NSDL National Science Digital Library

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

Steve Festin (Hamilton College;)



Meta-analysis of the effect of thiazolidinediones on serum C-reactive protein levels.  


We conducted a meta-analysis of randomized clinical trials to evaluate the effect of thiazolidinediones on serum C-reactive protein levels. Compared with placebo, treatment with thiazolidinediones significantly decreased the serum C-reactive protein levels (mean -0.82 mg/L, 95% confidence interval -1.15 to -0.49 mg/L, p <0.0001). In a subgroup analysis, the effect of thiazolidinediones on the serum C-reactive protein levels was more pronounced in diabetic patients (mean -1.24 mg/L, 95% confidence interval -2.15 to -0.32 mg/L, p = 0.008) compared with nondiabetic patients (mean -0.27 mg/L, 95% confidence interval -0.41 to -0.14 mg/L, p <0.0001). PMID:16490432

Qayyum, Rehan; Adomaityte, Jurga



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

PubMed Central

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

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



Gross protein influence upon blood plasma and serum self organization processes in patients with coronary heart disease  

NASA Astrophysics Data System (ADS)

Blood plasma pattern formation is a process sensitive to environment and carrier properties, and plasma biochemical content. 96 patients with coronary heart disease (CHD) were involved in the study. Control group include 12 practically health persons (PHP). Platelets poor plasma and serum were used to study functional morphology. Plasma and serum samples of equal volume were placed on degreased glass carrier with after going wedge dehydration. The result of wedge dehydration is a formation of a special structure called facia. To the samples of compare albumin solution was added. Morphology of prepared facies was studied by means of light microscopy ("Lomo Biolam P2-1") with 10 times magnification. All received facies were of the same principle structure with central, intermediate and edge zones. Zone index was increasing in samples with albumin adding. Special structures obligatory to atherosclerosis, vessels stiffness increase and hypoxia were found in facies of plasma and serum of patients with CHD. Quantity of these structures correlated to protein concentration (p = 0.021). Samples' drying period was also increasing in samples of compare, and differed significantly in patients with CHD and PHP. In our study gross proteins concentration increase modified plasma and serum morphology. Albumin solution can be proposed as a probe to elucidate differences of facies of patients with CHD and PHP.

Malinova, Lidia I.; Sergeeva, Uliya V.; Simonenko, Georgy V.; Denisova, Tatiana P.; Tuchin, Valery V.



Association of dehydroepiandrosterone sulfate, serum lipids, C-reactive protein and body mass index with age-related macular degeneration.  


The present study was designed to evaluate the associations between exudative age-related macular degeneration (AMD) and the serum concentrations of C-reactive protein (CRP), dehydroepiandrosterone sulfate (DHEAS), and lipids as well as the relationship between exudative AMD and body mass index (BMI). This cross-sectional study included of 141 healthy control subjects (70 males and 71 females with a mean age of 71.01 ± 3.84 years) and 142 exudative AMD patients (70 males and 72 females with a mean age of 70.92 ± 3.60 years). BMI and the serum concentrations of CRP, DHEAS, and lipids were measured in both groups. The data were statistically analysed using the Mann-Whitney U test, Chi squared test, independent sample t test, Cramer's V, point biserial correlation and logistic regression analysis. BMI values and serum concentrations of CRP, total cholesterol, and low-density lipoprotein (LDL) cholesterol were significantly higher in exudative AMD patients compared with normal controls (p values were 0.001, <0.001, 0.005 and <0.001, respectively). The serum concentrations of DHEAS were not significantly different between the controls and the exudative AMD patients for the subgroups of either gender (p values in males and females were 0.785 and 0.159, respectively). A logistic regression analysis revealed that the BMI and serum concentration of CRP moderately contributed to the predictive ability of the model (odds ratios were 1.205 and 1.179, respectively). Elevated total cholesterol concentrations and LDL cholesterol concentrations, BMI values and serum concentrations of CRP were associated with exudative AMD. However, no association between the serum DHEAS concentration and exudative AMD was established. PMID:23377999

Ula?, Fatih; Balbaba, Mehmet; Ozmen, Sedat; Celebi, Serdal; Do?an, Umit



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 ADA and C-reactive Protein in Rheumatoid Arthritis  

Microsoft Academic Search

Immunological and inflammatory reactions play a pivotal role in the initiation and perpetuation of rheumatoid arthritis. The present study is an attempt to estimate the levels of adenosine deaminase (ADA) activity, a marker for cell mediated immunity and C-reactive protein a marker for inflammation in patients with rheumatoid arthritis.75 cases presenting rheumatoid arthritis and same number of age and sex

H. Surekha Rani; G. Madhavi; B. M. V. Srikanth; P. Jharna; U. R. K. Rao; A. Jyothy



BPV E1 protein alters the kinetics of cell cycle entry of serum starved mouse fibroblasts  

SciTech Connect

A stable bovine papillomarvirus E1 expressing cell line (C2E1) was used to investigate the effects of E1 protein on the requirement for growth factors during serum-induced reentry from quiescence to proliferation. Flow cytometric bivariate DNA/PCNA analysis was utilized to study the expression of proliferating cell nuclear antigen (PCNA) concomitant with this transition. C2E1 cells, unlike the control cells (CNEO), were able to reenter the cell cycle when stimulated with low serum (1%). Stimulation with 10% serum revealed that C2E1 cells entered the first cell cycle faster than CNEO, indicating that E1 protein decreased the time of progression from GO state upon serum activation. It was also shown that PCNA expression started earlier in C2E1 cells than in CNEO cells after quiescent cells were stimulated with 10% serum. Addition of 1% serum was able to induce PCNA expression in C2E1 but not in CNEO cells in the first 24 h after stimulation. Using Triton X-100 treatment, it was found that the distribution between bound and unbound forms of PCNA was altered in E1-expressing cells compared to CNEO cells. Based on these results, it is suggested that E1 might possess mitogen-like properties. 30 refs., 4 figs.

Belyavskyi, M.; Miller, J.; Belyavskaya, E.; Wilson, V. [Texas A& M Univ., College Station, TX (United States)



A unique property of fetal bovine serum: high levels of protein-glutathione mixed disulfides.  


Fetal bovine serum has been reported to delay or inhibit "spontaneous" neoplastic transformation in vitro as compared with all other sera tested. The present results indicate that fetal bovine serum is also unique in containing high levels of protein-glutathione mixed disulfides (3 to 7 microng glutathione as mixed disulfide per ml serum). The level of mixed disulfide appears to vary in accordance with the period of gestation of the fetal calves used to prepare the serum, decreasing below detectable levels (less than 0.2 microng per ml) with near-term fetal calves. Calf, adult bovine, fetal horse, and swine sera did not contain detectable levels of this type of mixed disulfide. PMID:67079

Bump, E A; Reed, D J



Purification of lipopolysaccharide-binding protein from bovine serum  

Microsoft Academic Search

Lipopolysaccharide-binding protein (LBP) plays a central role in presentation of bacterial-derived lipopolysaccharide (LPS; endotoxin) to leukocytes such as macrophages and neutrophils. Interaction of LBP with LPS is significant because LBP-LPS complexes promote activation of leukocytes and the immune system, which results in enhanced secretion of a spectrum of proinflammatory cytokines. An improved, simplified method was used to purify bovine LBP

Philip N. Bochsler; Zhengang Yang; Charles L. Murphy; Roger C. Carroll



Prion protein detection in serum using micromechanical resonator arrays  

Microsoft Academic Search

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

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



Association of Serum Total Bilirubin with Serum High Sensitivity C-reactive Protein in Middle-aged Men  

PubMed Central

Background It has been suggested that bilirubin has an inverse association with cardiovascular disease (CVD) due to its antioxidant properties. However, there are few data regarding the relationship between serum total bilirubin (sTB) and risk factors for CVD in Koreans. This study aimed to evaluate the relationship between sTB and high sensitivity C-reactive protein (hsCRP), which is an independent risk factor for CVD. Methods We performed a cross sectional study in 6,800 men who were examined at a health promotion center at a university hospital in Korea between May 2005 and June 2006. We grouped the subjects according to values of serum hsCRP (above or below 1.0 mg/L) and compared the characteristics of the two groups. To evaluate the relationship between sTB and hsCRP, we classified the subjects according to quartile values of sTB. Multivariate logistic regression analyses were used to analyze the relationship of levels of sTB and hsCRP after adjusting for known risk factors for CVD. Results Serum hsCRP was significantly associated with body mass index (BMI), smoking, diabetes, hypertension, fasting plasma glucose, systolic blood pressure, alanine aminotransferase, and total cholesterol/high density lipoprotein (TC/HDL-C) ratio, but not with age or alcohol use. As levels of sTB increased, there was a decrease in age, numbers of smokers, BMI, and TC/HDL ratio. Compared to the lowest quartile of sTB, levels of hsCRP decreased with odds ratios of 0.82 (95% CI, 0.71 to 0.96), 0.75 (95% CI, 0.65 to 0.88), and 0.63 (95% CI, 0.54 to 0.74) in the 2nd, 3rd, and 4th quartiles of bilirubin, respectively. Conclusion Bilirubin may be inversely associated with hsCRP

Yu, Kiwoong; Sung, Eunju; Shin, Hocheol; Lee, Hyewon



Microassay for the quantitation of protein precipitable polyphenols: use of bovine serum albumin–benzidine conjugate as a protein probe  

Microsoft Academic Search

A simple, sensitive and indirect spectrophotometric method for the determination of protein precipitable polyphenols (tannins) has been developed, based on the ability of the polyphenols to precipitate the synthetic, brown coloured azo-protein, bovine serum albumin–benzidine conjugate (BSA–benzidine, mole ratio 1:7), which shows an absorption maxima at 405nm. The amount of unprecipitated BSA–benzidine is measured directly at 405nm, which is inversely

C. V Ratnavathi; R. B Sashidhar



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


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

Denner, Joachim



Serum resistance in Haemophilus parasuis SC096 strain requires outer membrane protein P2 expression.  


Haemophilus parasuis outer membrane protein P2 (OmpP2), the most abundant protein in the outer membrane, has been identified as an antigenic protein and a potential virulence factor. To study the precise function of OmpP2, an ompP2-deficient mutant (?ompP2) of a H. parasuis serovar 4 clinical strain SC096 was constructed by a modified natural transformation system. Compared with the wild-type SC096 strain, the ?ompP2 mutant showed a pronounced growth defect and exhibited significantly greater sensitivity to the bactericidal action of porcine and rabbit sera, whereas the complemented strain could restore the growth and serum resistance phenotypes. The results indicated that H. parasuis OmpP2 from SC096 strain is an important surface protein involved in serum resistance. PMID:22092746

Zhang, Bin; Feng, Saixiang; Xu, Chenggang; Zhou, Suming; He, Yanbing; Zhang, Lingyun; Zhang, Jianmin; Guo, Lili; Liao, Ming



Serum lipopolysaccharide-binding protein prediction of severe bacterial infection in cirrhotic patients with ascites.  


Serum lipopolysaccharide-binding protein is increased in a subset of non-infected ascitic cirrhotic patients, a finding previously related to bacterial passage from the gut to the circulation without overt infection. We prospectively analysed the risk factors associated with a first episode of severe bacterial infection in 84 ascitic cirrhotics, followed up for a median of 46 weeks. The cumulative probability of such infection in patients with raised and normal lipopolysaccharide-binding protein was 32.4% and 8.0% (p=0.004), respectively. Increased lipopolysaccharide-binding protein was the only factor independently associated with severe bacterial infection in a multivariate analysis (relative risk 4.49, 95% CI 1.42-14.1). Monitoring of serum lipopolysaccharide-binding protein could, therefore, help to target cirrhotic patients with ascites for antibiotic prophylaxis. PMID:15145636

Albillos, Agustín; de-la-Hera, Antonio; Alvarez-Mon, Melchor



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.



Integrative Proteomic Analysis of Serum and Peritoneal Fluids Helps Identify Proteins that Are Up-Regulated in Serum of Women with Ovarian Cancer  

PubMed Central

Background We used intensive modern proteomics approaches to identify predictive proteins in ovary cancer. We identify up-regulated proteins in both serum and peritoneal fluid. To evaluate the overall performance of the approach we track the behavior of 20 validated markers across these experiments. Methodology Mass spectrometry based quantitative proteomics following extensive protein fractionation was used to compare serum of women with serous ovarian cancer to healthy women and women with benign ovarian tumors. Quantitation was achieved by isotopically labeling cysteine amino acids. Label-free mass spectrometry was used to compare peritoneal fluid taken from women with serous ovarian cancer and those with benign tumors. All data were integrated and annotated based on whether the proteins have been previously validated using antibody-based assays. Findings We selected 54 quantified serum proteins and 358 peritoneal fluid proteins whose case-control differences exceeded a predefined threshold. Seventeen proteins were quantified in both materials and 14 are extracellular. Of 19 validated markers that were identified all were found in cancer peritoneal fluid and a subset of 7 were quantified in serum, with one of these proteins, IGFBP1, newly validated here. Conclusion Proteome profiling applied to symptomatic ovarian cancer cases identifies a large number of up-regulated serum proteins, many of which are or have been confirmed by immunoassays. The number of currently known validated markers is highest in peritoneal fluid, but they make up a higher percentage of the proteins observed in both serum and peritoneal fluid, suggesting that the 10 additional markers in this group may be high quality candidates.

Amon, Lynn M.; Law, Wendy; Fitzgibbon, Matthew P.; Gross, Jennifer A.; O'Briant, Kathy; Peterson, Amelia; Drescher, Charles; Martin, Daniel B.; McIntosh, Martin



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

PubMed Central

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

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



Grouped Variations in the Occurrence of New Protein Components in Normal Human Serum  

Microsoft Academic Search

A METHOD of zone electrophoresis using starch gel as the supporting medium has been developed. The advantages of zone electrophoresis and the convenience of protein detection by staining methods are combined with a resolving power in some cases superior to that obtained in free-solution boundary-electrophoresis. The method has been tested with human serum proteins, with cabbage enzyme preparations1, and with

O. Smithies



Serum electrolyte and protein modification during different workload in jumper horse  

Microsoft Academic Search

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

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



Temporal changes in serum concentrations of acute phase proteins in newborn dairy calves  

Microsoft Academic Search

Age-dependent changes in blood concentrations of four bovine acute phase proteins (APPs), serum amyloid A (SAA), lipopolysaccharide binding protein (LBP), haptoglobin (Hp) and alpha1-acid glycoprotein (AGP), were examined using two groups of newborn dairy calves. APP concentrations were monitored for either 3 weeks (Group A, n=13) or 2 months (Group B, n=13) after birth. Blood was collected at day 0–1

Toomas Orro; Stine Jacobsen; Jean-Philippe LePage; Theo Niewold; Sakari Alasuutari; Timo Soveri



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

Microsoft Academic Search

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

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



Effects of serum proteins on in vitro melamine-cyanurate crystal formation.  


Melamine toxicity is recognized as a distinct form of renal failure due to occlusion of the renal tubules by the compound melamine and its deaminated derivative, cyanuric acid. The morphology of melamine-cyanurate crystals in vivo differs from that in vitro, being rounded in the former case but needle-like in the latter. The reasons for this difference in morphology between in vivo and in vitro crystals remain unknown. In the present study, we investigated the in vitro effects of several possible intra-renal factors, i.e., pH and serum and urinary proteins, on the morphology of melamine-cyanurate crystals in order to clarify what might be responsible for the formation of rounded urolites in vivo. We found that serum proteins, such as fetal bovine serum, bovine serum albumin and bovine gamma-globulin, can alter the morphology of melamine cyanurate, turning it into rounded crystals. The urinary protein beta-2-microglobulin had a less pronounced effect. The crystal morphology was unaffected by pH. Based on the present in vitro findings and known clinical data, we suggest a putative protein-related model for melamine-cyanurate formation in the kidney. PMID:22813964

Taksinoros, Sarawut; Murata, Hideo



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

Technology Transfer Automated Retrieval System (TEKTRAN)

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


Identification of Patients With Head and Neck Cancer Using Serum Protein Profiles  

Microsoft Academic Search

Background: New and more consistent biomarkers of head and neck squamous cell carcinoma (HNSCC) are needed to improve early detection of disease and to moni- tor successful patient management. Objective: To determine if a new proteomic technol- ogy can correctly identify protein expression profiles for cancer in patient serum samples as well as detect the pres- ence of a known

J. Trad Wadsworth; Kenneth D. Somers; Brendan C. Stack; Lisa Cazares; Gunjan Malik; Bao-Ling Adam; George L. Wright; O. John Semmes



Serum cleaved tau protein levels and clinical outcome in adult patients with closed head injury  

Microsoft Academic Search

Study Objective: Intracranial injuries (ICI) are associated with high mortality and morbidity. Unfortunately, tools for diagnosis and risk stratification of ICIs are limited in the emergency department setting. We determine the relationship between the presence or absence of a detectable cleaved serum tau protein (?c), ICI, and outcome at hospital discharge in adults with closed head injuries (CHI). Methods: This

George J. Shaw; Edward C. Jauch; Frank P. Zemlan



Comparison of Bromcresol Green and Agarose Protein Electrophoresis for Quantitation of Serum Albumin in Multiple Myeloma  

Microsoft Academic Search

Background: The International Staging System for mul- tiple myeloma has increased the importance of accurate measurement of serum albumin. Two common albumin assays, bromcresol green (BCG) and agarose gel protein electrophoresis (PEL), frequently yield discordant re- sults, creating confusion regarding which assay is supe- rior for use in myeloma. Methods: We measured albumin by BCG on a Roche Modular system,

Christine L. H. Snozek; Amy K. Saenger; Philip R. Greipp; Sandra C. Bryant; Robert A. Kyle; S. Vincent Rajkumar; Jerry A. Katzmann



Serum proteins and nutritional status of free-living Thai elderly  

Microsoft Academic Search

Fifty-six Thai males and 146 Thai females aged 60 years and above visiting a special clinic for the elderly were investigated. The serum protein and immunoglobulin of these elderly were assessed. Anthropometric measurements were also taken. From a random sample of the group, the dietary intake of main nutrients was determined by a 24 h recall method with the help

Praneet Pongpaew; Rungsunn Tungtrongchitr; Benjaluck Phonrat; Venus Supawan; Areeya Lertchawanakul; Somsak Tawprasert; Niyomsri Vudhivai; Frank-Peter Schelp



Acute phase protein quantitation in serum samples from healthy Atlantic bottlenose dolphins (Tursiops truncatus).  


Acute phase proteins (APPs) have been studied in many companion and large animals and have been reported to have a differential sensitivity to traditional markers of inflammation. Studies have been performed indicating the conservation of these proteins as well as the application and cross-reactivity of automated assays among different species, but few reports have detailed APPs in marine mammal species. In the present study, automated assays were utilized to generate reference intervals for C-reactive protein, haptoglobin, and serum amyloid A using 44 serum samples from healthy Atlantic bottlenose dolphins (Tursiops truncatus). A total of 25 samples were obtained from dolphins under human care and 19 samples were obtained from free-ranging dolphins. Mild yet statistically significant differences were observed in levels of haptoglobin and serum amyloid A between these groups. The reference intervals from the combined groups were as follows: C-reactive protein 3.1-19.7 mg/l, haptoglobin 0-0.37 mg/ml, and serum amyloid A 17.5-42.9 mg/l. These baseline data should provide an important foundation for future studies of the application of APP quantitation in monitoring the health and stressors of dolphins under human care and with live capture of free-ranging dolphins. PMID:23242666

Cray, Carolyn; Arheart, Kristopher L; Hunt, Michael; Clauss, Tonya; Leppert, Lynda L; Roberts, Kevin; McCulloch, Stephen D; Goldstein, Juli D; Gonzalez, Christie; Sweeney, Jay; Stone, Rae; Fair, Patricia A; Bossart, Gregory D



Soy protein, casein, and zein regulate histidase gene expression by modulating serum glucagon.  


Glucagon has been postulated as an important physiological regulator of histidase (Hal) gene expression; however, it has not been demonstrated whether serum glucagon concentration is associated with the type and amount of protein ingested. The purpose of the present work was to study the association between hepatic Hal activity and mRNA concentration in rats fed 18 or 50% casein, isolated soy protein, or zein diets in a restricted schedule of 6 h for 10 days, and plasma glucagon and insulin concentrations. On day 10, five rats of each group were killed at 0900 (fasting), and then five rats were killed after being given the experimental diet for 1 h (1000). Rats fed 50% casein or soy diets showed higher Hal activity than the other groups studied. Rats fed 50% zein diets had higher Hal activity than rats fed 18% casein, soy, or zein diets, but lower activity than rats fed 50% casein or soy diets. Hal mRNA concentration followed a similar pattern. Hal activity showed a significant association with serum concentrations of glucagon. Serum glucagon concentration was significantly correlated with protein intake. Thus the type and amount of protein consumed affect Hal activity and expression through changes in serum glucagon concentrations. PMID:12376330

Tovar, Armando R; Ascencio, Claudia; Torres, Nimbe



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

Microsoft Academic Search

Skeletal myogenesis is precisely regulated by multiple transcription factors. Previously, we demonstrated that enhancer of polycomb 1 (Epc1) induces skeletal muscle differentiation by potentiating serum response factor (SRF)-dependent muscle gene activation. Here, we report that an interacting partner of Epc1, ret finger protein (RFP), blocks skeletal muscle differentiation. Our findings show that RFP was highly expressed in skeletal muscles and

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



Serum cytokine profiles associated with specific adjuvants used in a DNA prime-protein boost vaccination strategy.  


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. PMID:24019983

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



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



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.



Identification of the Major Mannose-Binding Proteins from Chicken Egg Yolk and Chicken Serum as Immunoglobulins  

Microsoft Academic Search

Chicken egg yolk contains a mannose-binding protein that could be purified with a modification of the procedure of affinity chromatography and gel filtration used for chicken serum mannose-binding protein. The yolk protein was indistinguishable from the serum protein with respect to apparent molecular masses (169 ± 7 kDa), subunits (74 kDa and 27 kDa, in approximately 1:1 ratio) produced after

Ke-Yi Wang; Craig A. Hoppe; Pradip K. Datta; Amy Fogelstrom; Yuan Chuan Lee



Antigenicity, storage, and aging: physiologic autoantibodies to cell membrane and serum proteins and the senescent cell antigen  

Microsoft Academic Search

Normal human serum contains autoantibodies to a wide range of cellular and serum proteins. IgG autoantibodies to cell membrane proteins spectrin, syndein (Band 2.1), Band 3 degradation products, and the senescent cell antigen are among them. Physiologic autoantibodies to the senescent cell antigen, a ~62 000 dalton glycopeptide derived from Band 3, initiate removal of senescent, damaged, and stored cells

M. M. B. Kay; K. Sorensen; P. Wong; P. Bolton



Preliminary proteomic-based identification of a novel protein for Down's syndrome in maternal serum.  


Prenatal screening for Down's syndrome (DS) is in need of improvement. As a powerful platform, proteomics techniques could also be used for identification of new biomarkers for DS screening. In this case-control proteome study, pregnant women were diagnosed prenatally by karyotype analysis from amniotic fluid (AF). Maternal serum samples were collected from six pregnancies with fetuses affected by DS and six pregnancies with normal fetuses. First, we used two-dimensional electrophoresis and mass spectrometry to identify the different levels of expression of proteins in maternal serum between the DS and control groups in the second trimester. Second, we used bioinformatics to analyze the proteins by DAVID. Then, the interesting candidates were further tested by enzyme-linked immunosorbent assay (ELISA). Twenty-nine proteins were successfully identified in maternal serum obtained from pregnancies with fetuses affected by DS. The top five proteins up-regulated were serotransferrin (TF), alpha-1b-glycoprotein (A1BG), desmin (DES), alpha-1-antitrypsin (SERPINA1) and ceruloplasmin (CP), while serum amyloid P-component (APCS) was the most down-regulated protein. These 29 proteins were categorized based on binding, catalytic activity and enzyme regulator activity. The biological roles were involved in biological regulation, metabolic processes, cellular processes and response to a stimulus. Based on ELISA, the median concentrations of CP and complement factor B (CFB) were 332.3 and 412.3 ng/mL, respectively. The concentrations of CP and CFB were significantly higher in the DS group than in the control group (P < 0.05). In conclusion, proteomic approaches offer the possibility of further improving the performance of DS screening and our identification of up- and down-regulated proteins may lead to new candidates for DS screening. PMID:22678011

Yu, Bin; Zhang, Bin; Wang, Jing; Wang, Qiu-wei; Huang, Rui-ping; Yang, Yu-qi; Shao, Shi-he



Transient increase of serum Clara cell protein (CC16) after exposure to smoke.  

PubMed Central

OBJECTIVES: Smoke inhalation is a well known cause of airways injury in firefighting personnel. The aim of this study was to evaluate whether toxic effects of smoke on the respiratory tract can be detected by measuring Clara cell protein (CC16), a recently described serum marker of lung function. METHODS: CC16 was measured by a sensitive latex immunoassay in the serum of six voluntary firefighters from a chemical plant who had inhaled smoke from the combustion of polypropylene for about 20 minutes. The protein was measured immediately after the fire and 10 days later. The values were compared with those of six control workers examined simultaneously. RESULTS: The mean (SD) concentration of CC16 in the serum of firefighters after the fire (54.4 (34.9) micrograms/l) was significantly higher than that of controls (19.5 (11.7), P = 0.04). 10 days later, serum CC16 from firefighters had returned to the concentrations found in controls (15.9 (2.76) v 17.7 (12.5)). With the values at day 10 as a baseline, the rise of serum CC16 was estimated at 328% on average (range 100%-564%). These changes were found in the absence of any functional sign of lung impairment. CONCLUSION: Acute exposure to smoke results in a transient increase of CC16 in serum due most likely to an increased permeability of the bronchoalveolar/blood barrier. Serum CC16 seems potentially to be a new biomarker for the early detection of acute airways injury caused by smoke.

Bernard, A; Hermans, C; Van Houte, G



Levels of three inflammation markers, C-reactive protein, serum amyloid A protein and procalcitonin, in the serum and cerebrospinal fluid of patients with meningitis.  


The levels of C-reactive protein (CRP) and serum amyloid A protein (SAA) in blood are increased in patients with inflammatory diseases as acute phase proteins. Most of the presently used indicators of inflammation, such as body temperature, white cell count, erythrocyte sedimentation rate or CRP, are non-specific parameters. In contrast, procalcitonin (PCT) has been reported to be selectively induced by severe bacterial infection during the systemic inflammatory response syndrome (SIRS), and also in sepsis or multiorgan dysfunction syndrome. PCT expression is only slightly induced, if at all, by viral infections, autoimmune disorders, neoplastic diseases and trauma of surgical intervention. We measured the concentrations of CRP, SAA and PCT in the sera and cerebrospinal fluid (CSF) of 30 patients with bacterial, viral, or mycotic meningitis, and 12 patients with a noninflammatory central nervous system disease as controls. An extremely high CRP level in CSF of above 100 microg/L was seen in all seven bacterial meningitis patients and in only 10% of the viral meningitis patients. A high SAA level in CSF of greater than 10 microg/L was observed in all of the bacterial meningitis and mycotic meningitis patients, and in 95% of the viral meningitis patients. Among those with bacterial meningitis, the serum PCT level was more elevated in those with more serious bacterial meningitis. The PCT level in the CSF did not significantly differ among the patients with the three types of meningitis. However, the serum PCT level was very high above 0.1 microg/L in all seven bacterial meningitis patients, especially in the clinically serious cases. PMID:11763415

Shimetani, N; Shimetani, K; Mori, M



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

PubMed Central

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

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



Soluble CD86 protein in serum samples of patients with asthma  

PubMed Central

Background: Previous studies have reported that soluble (s) CD86 is involved in the initiation of the immune response. A study was undertaken to investigate the concentrations of sCD86 in serum samples from patients with bronchial asthma and to determine the cell origin of sCD86. Methods: Serum sCD86 concentrations were measured in 52 asthmatic subjects and 25 non-atopic normal volunteers using an enzyme linked immunosorbent assay, and the relationship of serum sCD86 concentrations to asthma severity and to total and differential white cell counts was analysed. Each type of white blood cell was purified and cultured in vitro to determine the cell origin of serum sCD86. Results: Serum samples from patients with an acute asthma exacerbation had much higher levels of sCD86 (585.4 (20.5) IU/ml) than those from stable asthmatics (479.6 (15.7) IU/ml, p<0.001) and healthy individuals (435.1 (13.8) IU/ml, p<0.001), and there was no difference between the latter two groups (p = 0.079). In asthmatic subjects the serum sCD86 level was inversely correlated with airway responsiveness, forced expiratory volume in 1 second, and with arterial carbon dioxide tension. In addition, the serum sCD86 level was positively correlated with numbers of lymphocytes, eosinophils, monocytes, but not neutrophils. The in vitro experiments indicated that sCD86 was produced by monocytes. Conclusions: The serum sCD86 protein level was significantly increased in asthmatic subjects during an exacerbation and correlated with the severity of asthma. sCD86 is most probably derived from monocytes in the peripheral blood.

Shi, H; Xie, Z; Deng, J; Chen, Y; Xiao, C



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:



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

PubMed Central

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

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



Serum C-reactive protein: a prognostic factor in metastatic urothelial cancer of the bladder.  


Until today, there is no reliable prognostic or predictive parameter for the prognosis of patients with metastatic urothelial cancer of the bladder prior to chemotherapy. Recently, serum C-reactive protein (CRP) level has been shown to be associated with survival of patients with various malignancies including localized and metastatic renal cell carcinoma, upper urinary tract as well as penile cancer. The aim of this study was to evaluate the prognostic impact of the pretreatment CRP serum level in patients with metastatic urothelial cancer of the bladder. We retrospectively evaluated 34 patients with metastatic urothelial cancer of the bladder and information about the CRP level prior to chemotherapy. The CRP level was correlated with patient- and tumor-specific characteristics. Kaplan-Meier and log-rank analyses were employed to calculate progression-free (PFS) and overall survival (OS). Receiver operating characteristics (ROC) analysis was used to determine an optimal prognostic CRP cutoff value to predict cancer-specific death. The median PFS to first-line chemotherapy and the OS for the whole cohort were 3.3 and 24.3 months, respectively. Serum CRP in mg/l was significantly associated with patients' survival (HR 1.02, p < 0.001, univariate Cox-regression). ROC analysis identified a CRP value of 80 mg/l to be the optimal cutoff. The median PFS was 4.5 and 3.0 months (p = 0.08; Mann-Whitney test), and the calculated 1-year OS was 82.6 and 22.2 % for patients with a CRP <80 and ?80 mg/l, respectively (log-rank, p < 0.001). In contrast, neither T-stage, tumor grade, sex, age nor the body mass index was related to the CRP level or associated with overall survival. This is the first analysis revealing that the CRP value prior to systemic treatment might be of prognostic significance and could enable better risk stratification for patients with metastatic urothelial cancer of the bladder. PMID:24005810

Eggers, Hendrik; Seidel, Christoph; Schrader, Andres Jan; Lehmann, Rieke; Wegener, Gerd; Kuczyk, Markus A; Steffens, Sandra



Association of Serum Ceruloplasmin Level with Obesity: Some Components of Metabolic Syndrome and High-Sensitive C-Reactive Protein in Iran  

PubMed Central

Background. One of the mechanisms that has been suggested for obesity related metabolic disturbances is obesity-induced inflammation. Pro-inflammatory cytokines generated in adipose tissue can increase hepatic synthesis of inflammation-sensitive plasma proteins (ISPs) including ceruloplasmin (Cp). In this study we aimed to investigate the relation between serum Cp level and obesity. Methods. 61 persons with body mass index (BMI) ? 25?kg/m2 (case group) and 61 persons with BMI < 25?kg/m2 (control group) were included in this study with a case-control design. Serum Cp levels, triglyceride level, fating blood glucose, total cholesterol, LDL-cholesterol, HDL-cholesterol and hsCRP were measured in both groups. Results. We did not observe any significant association between serum Cp level and BMI in all subjects [OR: 1.02 (CI, 0.967 to 1.07)] and in case (? = 0.012, P = 0.86) and control groups (? = 0.49, P = 0.07) separately. However, in control group, this positive association was marginally significant. We found a positive correlation between serum Cp level and serum triglyceride level. Conclusion. Serum Cp level was not related to obesity in this group of subjects. None of the baseline variables could predict obesity in this group of subjects, including serum Cp level, FBS, total cholesterol, LDL and HDL- cholesterols and hsCRP.

Safavi, Seyyed Morteza; Ziaei, Rahele; Maracy, Mohammad Reza



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



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

Microsoft Academic Search

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




Serum heat shock protein 70 level as a biomarker of exceptional longevity.  


Heat shock proteins are highly conserved proteins that, when produced intracellularly, protect stress exposed cells. In contrast, extracellular heat shock protein 70 (Hsp70) has been shown to have both protective and deleterious effects. In this study, we assessed heat shock protein 70 for its potential role in human longevity. Because of the importance of HSP to disease processes, cellular protection, and inflammation, we hypothesized that: (1) Hsp70 levels in centenarians and centenarian offspring are different from controls and (2) alleles in genes associated with Hsp70 explain these differences. In this cross-sectional study, we assessed serum Hsp70 levels from participants enrolled in either the New England Centenarian Study (NECS) or the Longevity Genes Project (LGP): 87 centenarians (from LGP), 93 centenarian offspring (from NECS), and 126 controls (43 from NECS, 83 from LGP). We also examined genotypic and allelic frequencies of polymorphisms in HSP70-A1A and HSP70-A1B in 347 centenarians (266 from the NECS, 81 from the LGP), 260 NECS centenarian offspring, and 238 controls (NECS: 53 spousal controls and 106 septuagenarian offspring controls; LGP: 79 spousal controls). The adjusted mean serum Hsp70 levels (ng/mL) for the NECS centenarian offspring, LGP centenarians, LGP spousal controls, and NECS controls were 1.05, 1.13, 3.07, 6.93, respectively, suggesting that a low serum Hsp70 level is associated with longevity; however, no genetic associations were found with two SNPs within two hsp70 genes. PMID:17027907

Terry, Dellara F; Wyszynski, Diego F; Nolan, Vikki G; Atzmon, Gil; Schoenhofen, Emily A; Pennington, JaeMi Y; Andersen, Stacy L; Wilcox, Marsha A; Farrer, Lindsay A; Barzilai, Nir; Baldwin, Clinton T; Asea, Alexzander



The small GTP-binding protein Rho links G protein-coupled receptors and G?12 to the serum response element and to cellular transformation  

PubMed Central

Receptors coupled to heterotrimeric G proteins can effectively stimulate growth promoting pathways in a large variety of cell types, and if persistently activated, these receptors can also behave as dominant-acting oncoproteins. Consistently, activating mutations for G proteins of the G?s and G?i2 families were found in human tumors; and members of the G?q and G?12 families are fully transforming when expressed in murine fibroblasts. In an effort aimed to elucidate the molecular events involved in proliferative signaling through heterotrimeric G proteins we have focused recently on gene expression regulation. Using NIH 3T3 fibroblasts expressing m1 muscarinic acetylcholine receptors as a model system, we have observed that activation of this transforming G protein-coupled receptors induces the rapid expression of a variety of early responsive genes, including the c-fos protooncogene. One of the c-fos promoter elements, the serum response element (SRE), plays a central regulatory role, and activation of SRE-dependent transcription has been found to be regulated by several proteins, including the serum response factor and the ternary complex factor. With the aid of reporter plasmids for gene expression, we observed here that stimulation of m1 muscarinic acetylcholine receptors potently induced SRE-driven reporter gene activity in NIH 3T3 cells. In these cells, only the G?12 family of heterotrimeric G protein ? subunits strongly induced the SRE, while G?1?2 dimers activated SRE to a more limited extent. Furthermore, our study provides strong evidence that m1, G?12 and the small GTP-binding protein RhoA are components of a novel signal transduction pathway that leads to the ternary complex factor-independent transcriptional activation of the SRE and to cellular transformation.

Fromm, Christian; Coso, Omar A.; Montaner, Silvia; Xu, Ningzhi; Gutkind, J. Silvio



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

NSDL National Science Digital Library

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

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



Serum vascular adhesion protein-1 level is higher in smokers than non-smokers.  


Abstract Background: Semicarbazide-sensitive amine oxidase (SSAO)/vascular adhesion protein-1 (VAP-1) is involved in the pathogenesis of both atherosclerosis and cancer. Because chemical components and metabolites of cigarettes are deaminated by SSAO, the relationship between smoking and serum SSAO/VAP-1 was studied in humans. Methods: A total of 451 non-diabetic and normoalbuminuric Han Chinese subjects were recruited to participate in this study. Smoking history was obtained by using a questionnaire and those who smoked more than 100 cigarettes during a 6-month period were considered smokers. Serum VAP-1 concentration was measured by time-resolved immunofluorometric assay. Age, gender, waist circumference and estimated glomerular filtration rate (GFR) were adjusted in different statistical models. Results: Smokers were mainly male (85.7% versus 26.3%) and were more obese than non-smokers (p?serum VAP-1 concentrations were older (p?Serum VAP-1 concentration was higher in smokers than in non-smokers (p?serum VAP-1 concentration. Whether VAP-1 and its SSAO activity link the relationship between cigarette smoking, atherosclerosis and cancer requires further investigation. PMID:23802578

Wang, Yi-Chia; Li, Hung-Yuan; Wei, Jung-Nan; Lin, Mao-Shin; Shih, Shyang-Rong; Hua, Cyue-Huei; Smith, David J; Vanio, Jani; Chuang, Lee-Ming




PubMed Central

Human serum albumin (HSA) was explored for use as a stationary phase and ligand in affinity microcolumns for the ultrafast extraction of free drug fractions and the use of this information for the analysis of drug-protein binding. Warfarin, imipramine, and ibuprofen were used as model analytes in this study. It was found that greater than 95% extraction of all these drugs could be achieved in as little as 250 ms on HSA microcolumns. The retained drug fraction was then eluted from the same column under isocratic conditions, giving elution in less than 40 s when working at 4.5 mL/min. The chromatographic behavior of this system gave a good fit with that predicted by computer simulations based on a reversible, saturable model for the binding of an injected drug with immobilized HSA. The free fractions measured by this method were found to be comparable to those determined by ultrafiltration, and equilibrium constants estimated by this approach gave good agreement with literature values. Advantages of this method include its speed and the relatively low cost of microcolumns that contain HSA. The ability of HSA to bind many types of drugs also creates the possibility of using the same affinity microcolumn to study and measure the free fractions for a variety of pharmaceutical agents. These properties make this technique appealing for use in drug binding studies and in the high-throughput screening of new drug candidates.

Mallik, Rangan; Yoo, Michelle J.; Briscoe, Chad J.; Hage, David S.



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



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


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. PMID:22781126

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



Association of canine obesity with reduced serum levels of C-reactive protein.  


The prevalence of obesity is increasing in dogs as well as in humans. C-reactive protein (CRP) is an important tool for the detection of inflammation and/or early tissue damage and is linked to obesity in humans. The objective of the present study was to determine if serum CRP levels are altered in obese dogs. Fifteen lean (control group) and 16 overweight (obese group) dogs were examined. Blood samples were collected under fasted conditions for serum determination of CRP, glucose, insulin, cholesterol, triglyceride, and fructosamine. Results indicated that obese dogs were insulin resistant because serum insulin and insulin/glucose ratios were higher than in lean dogs (P < or = 0.05). Serum CRP concentrations were lower in obese dogs than in controls (P < or = 0.001). C-reactive protein was negatively correlated with insulin/glucose ratio (R = -0.42) and cholesterol (R = -0.39; P < or = 0.05). Furthermore, levels of cholesterol, triglycerides, and fructosamine were increased in the obese group compared with the control group. Based on these results, it can be postulated that CRP production is inhibited by obesity and insulin resistance in dogs. PMID:18319438

Veiga, Angela P M; Price, Christopher A; de Oliveira, Simone T; Dos Santos, Andréa P; Campos, Rómulo; Barbosa, Patricia R; González, Félix H D



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



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



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



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

PubMed Central

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

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



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


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

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



Fast human serum profiling through chemical depletion coupled to gold-nanoparticle-assisted protein separation.  


The use of chemical protein depletion in conjunction with gold-based nanoparticles for fast matrix assisted laser desoption ionization time of flight mass spectrometry-based human serum profiling was assessed. The following variables influencing the process were optimized: (i) amount of nanoparticles, (ii) sample pH, (iii) amount of protein and (iv) incubation time. pH was found the most important factor to be controlled, with an optimum range comprised between 5.8 and 6.4. The minimum incubation time to obtain an adequate profiling was 30 min. Using this approach, serum from five patients with lymphoma, five patients with myeloma and from two healthy volunteers were correctly classified using Principal component analysis. PMID:23141332

López-Cortés, Rubén; Oliveira, Elisabete; Núńez, Cristina; Lodeiro, Carlos; Páez de la Cadena, María; Fdez-Riverola, Florentino; López-Fernández, H; Reboiro-Jato, Miguel; Glez-Peńa, Daniel; Luis Capelo, José; Santos, Hugo M



Maternal Milk and Serum Vitamin B12, Folic Acid, and Protein Levels in Indian Subjects  

PubMed Central

Serum vitamin B12 and folic acid activity levels were studied in 47 mothers. The mean serum vitamin B12 value in non-vegetarian mothers was 228 ??g./ml. ± 38·9 SE, as compared to mean values 97 ??g./ml. ± 20·7 SE in lacto-vegetarians. There was no significant difference in folic acid levels, haemoglobin, and total proteins among the different dietetic groups. Analysis of milk revealed mean vitamin B12 values of 103 ??g./ml. in non-vegetarians as compared to 91 ??g./ml. in lacto-vegetarians; the difference was not statistically significant. Mean total protein and folic acid levels for milk were similar in different dietetic groups. The figures obtained for vitamin B12 content of human milk in these mothers are considerably lower than those reported in similar studies from Western countries.

Jathar, V. S.; Kamath, S. A.; Parikh, M. N.; Rege, D. V.; Satoskar, R. S.



[Effect of radiomimetics on the composition of proteins in the serum of dog].  


In this paper the protein serum of dogs was investigated after an application of the radiomimetics of beta, beta'-dichlorodiethyl sulphide by means of immunoelectrophoresis. The applied dose of beta, beta'-dichlorodiethyl sulphide is 35 mg of live weight s.c. per dog. It was found that together with the lengthened time after the application the precipitation lines in the sphere of beta1- and alpha2-globulins increased. The disintegration and distance of the precipitation lines in the direction of the electrophoretic migration and immunodiffusion. In our case SE and IE in agar provide only a rough semi-quantitative picture of the different protein fractions of dog serum and of their changes after an application of radiomimetics. PMID:818774

Hrus?ovský, J



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


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

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



Development of a high-throughput LC/APCI-MS method for the determination of thirteen phytoestrogens including gut microbial metabolites in human urine and serum.  


The investigation into the potential usefulness of phytoestrogens in the treatment of menopausal symptoms requires large-scale clinical trials that involve rapid, validated assays for the characterization and quantification of the phytoestrogenic precursors and their metabolites in biological matrices, as large interindividual differences in metabolism and bioavailability have been reported. Consequently, a new sensitive high-performance liquid chromatography-mass spectrometry method (HPLC-MS) for the quantitative determination of thirteen phytoestrogens including their most important gut microbial metabolites (genistein, daidzein, equol, dihydrodaidzein, O-desmethylangolensin, coumestrol, secoisolariciresinol, matairesinol, enterodiol, enterolactone, isoxanthohumol, xanthohumol and 8-prenylnaringenin) in human urine and serum within one single analytical run was developed. The method uses a simple sample preparation procedure consisting of enzymatic deconjugation followed by liquid-liquid extraction (LLE) or solid-phase extraction (SPE) for urine or serum, respectively. The phytoestrogens and their metabolites are detected with a single quadrupole mass spectrometer using atmospheric pressure chemical ionization (APCI), operating both in the positive and the negative mode. This bioanalytical method has been fully validated and proved to allow an accurate and precise quantification of the targeted phytoestrogens and their metabolites covering the lower parts-per-billion range for the measurement of relevant urine and serum levels following ingestion of phytoestrogen-rich dietary supplements. PMID:20299290

Wyns, Ciska; Bolca, Selin; De Keukeleire, Denis; Heyerick, Arne



The human serum proteome: display of nearly 3700 chromatographically separated protein spots on two-dimensional electrophoresis gels and identification of 325 distinct proteins.  


Plasma, the soluble component of the human blood, is believed to harbor thousands of distinct proteins, which originate from a variety of cells and tissues through either active secretion or leakage from blood cells or tissues. The dynamic range of plasma protein concentrations comprises at least nine orders of magnitude. Proteins involved in coagulation, immune defense, small molecule transport, and protease inhibition, many of them present in high abundance in this body fluid, have been functionally characterized and associated with disease processes. For example, protein sequence mutations in coagulation factors cause various serious disease states. Diagnosing and monitoring such diseases in blood plasma of affected individuals has typically been conducted by use of enzyme-linked immunosorbent assays, which using a specific antibody quantitatively measure only the affected protein in the tested plasma samples. The discovery of protein biomarkers in plasma for diseases with no known correlations to genetic mutations is challenging. It requires a highly parallel display and quantitation strategy for proteins. We fractionated blood serum proteins prior to display on two-dimensional electrophoresis (2-DE) gels using immunoaffinity chromatography to remove the most abundant serum proteins, followed by sequential anion-exchange and size-exclusion chromatography. Serum proteins from 74 fractions were displayed on 2-DE gels. This approach succeeded in resolving approximately 3700 distinct protein spots, many of them post-translationally modified variants of plasma proteins. About 1800 distinct serum protein spots were identified by mass spectrometry. They collapsed into 325 distinct proteins, after sequence homology and similarity searches were carried out to eliminate redundant protein annotations. Although a relatively insensitive dye, Coomassie Brilliant Blue G-250, was used to visualize protein spots, several proteins known to be present in serum in < 10 ng/mL concentrations were identified such as interleukin-6, cathepsins, and peptide hormones. Considering that our strategy allows highly parallel protein quantitation on 2-DE gels, it holds promise to accelerate the discovery of novel serum protein biomarkers. PMID:12872236

Pieper, Rembert; Gatlin, Christine L; Makusky, Anthony J; Russo, Paul S; Schatz, Courtney R; Miller, Stanton S; Su, Qin; McGrath, Andrew M; Estock, Marla A; Parmar, Prashanth P; Zhao, Ming; Huang, Shih-Ting; Zhou, Jeff; Wang, Fang; Esquer-Blasco, Ricardo; Anderson, N Leigh; Taylor, John; Steiner, Sandra



Does binding of complement factor H to the meningococcal vaccine antigen, factor H binding protein, decrease protective serum antibody responses?  


Factor H binding protein (fHbp) is a principal antigen in a multicomponent meningococcal vaccine recently licensed in Europe for prevention of serogroup B diseases. The protein recruits the complement downregulator, factor H (fH), to the bacterial surface, which enables the organism to resist complement-mediated bacteriolysis. Binding is specific for human fH. In preclinical studies, mice and rabbits immunized with fHbp vaccines developed serum bactericidal antibody responses, which in humans predict protection against developing meningococcal disease. These studies, however, were in animals whose fH did not bind to the vaccine antigen. Here we review the immunogenicity of fHbp vaccines in human fH transgenic mice. The data suggest that animals with high serum human fH concentrations have impaired protective antibody responses. Further, mutant fHbp vaccines with single amino acid substitutions that decrease fH binding are superior immunogens, possibly by unmasking epitopes in the fH binding site that are important for eliciting serum bactericidal antibody responses. Humans immunized with fHbp vaccines develop serum bactericidal antibody, but achieving broad coverage in infants required incorporation of additional antigens, including outer membrane vesicles, which increased rates of fever and local reactions at the injection site. The experimental results in transgenic mice predict that fHbp immunogenicity can be improved in humans by using mutant fHbp vaccines with decreased fH binding. These results have important public health implications for developing improved fHbp vaccines for control of serogroup B meningococcal disease and for development of vaccines against other microbes that bind host molecules. PMID:23740919

Granoff, Dan M; Ram, Sanjay; Beernink, Peter T



Changes in serum C-reactive protein levels in dogs with various disorders and surgical traumas  

Microsoft Academic Search

The serum levels of C-reactive protein (CRP) produced as an inflammatory response in dogs with various disorders and surgical traumas were measured by enzyme-linked immunoabsorbent assay and slide reversed passive latex agglutination test (RPLA). The CRP levels were greatly increased 1–2 days after surgery in most of the dogs (n=29) subjected to surgery. These levels had markedly decreased by the

S. Yamamoto; T. Shida; S. Miyaji; H. Santsuka; H. Fujise; K. Mukawa; E. Furukawa; T. Nagae; M. Naiki



Serum lipase, C-reactive protein, and interleukin-6 levels in ERCP-induced pancreatitis  

Microsoft Academic Search

Background: C-reactive protein (CRP) and interleukin-6 (IL-6) are elevated in acute pancreatitis. Limited studies have evaluated their role in ERCP-induced pancreatitis. The aim of this study was to assess the role of serum lipase, CRP, and IL-6 in ERCP-induced pancreatitis. Methods: Eighty-five patients (62 women, 23 men; mean age 43 years; range 16-85 years) who underwent ERCP were entered in

Madhukar Kaw; Sandeep Singh



Measurement of serum C-reactive protein concentration in myocardial ischaemia and infarction  

Microsoft Academic Search

Serum C-reactive protein (CRP) and creatine kinase (CK) MB levels were measured prospectively in patients with definite myocardial infarction, patients with spontaneous or exercise-induced angina, subjects undergoing coronary arteriography, and patients with non-cardiac chest pain. All individuals with infarction developed raised CRP levels and there was a significant correlation between the peak CRP and CK MB values. The CRP, however,

F C de Beer; C R Hind; K M Fox; R M Allan; A Maseri; M B Pepys



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

Microsoft Academic Search

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

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



An evaluation of metal bioaccessibility in estuarine sediments using the commercially available protein, bovine serum albumin  

Microsoft Academic Search

The bioaccessibility of metals (Al, Ca, Fe, Mn, Ag, Cd, Co, Cu, Ni, Pb, Sn, Zn) in oxic estuarine sediments has been evaluated using solutions of a commercially available protein (bovine serum albumin; BSA) that mimic the chemical conditions encountered in the gut environment of many deposit-feeding organisms. Over a 20 h incubation period with 5 g L?1 BSA, metal mobilisation was

Judit Kalman; Andrew Turner



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


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

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



Seasonal variations in serum protein fractions of dairy cows during different physiological phases  

Microsoft Academic Search

The seasonal variations of the level of serum total proteins and their electrophoretic fractions in dairy cows, living in\\u000a the Mediterranean area, were investigated. During different physiological phases, ten dairy cows were housed in a sheltered\\u000a pen in Sicily (Italy). Throughout the experimental period, ambient temperature and relative humidity were continuously recorded\\u000a with a data logger and temperature humidity index

Giuseppe Piccione; Vanessa Messina; Daniela Alberghina; Claudia Giannetto; Stefania Casella; Anna Assenza


Distinct roles of haptoglobin-related protein and apolipoprotein LI in trypanolysis by human serum  

Microsoft Academic Search

Apolipoprotein L-I (apoL-I) is a human high-density lipoprotein (HDL) component able to kill Trypanosoma brucei brucei by forming anion-selective pores in the lysosomal membrane of the parasite. Another HDL component, haptoglobin-related protein (Hpr), has been suggested as an additional toxin required for full trypanolytic activity of normal human serum. We recently reported the case of a human lacking apoL-I $({\\\\rm

Benoit Vanhollebeke; M. J. Nielsen; Yoshihisa Watanabe; Philippe Truc; Luc Vanhamme; Kazunori Nakajima; S. K. Moestrup; Etienne Pays



Serum C-Reactive Protein and Cognitive Function in Healthy Elderly Italian Community Dwellers  

Microsoft Academic Search

Background. Chronic low-grade inflammation, as measured with the peripheral serum marker C-reactive protein (sCRP), may be a risk factor for dementia in elderly persons. Methods. The relationship between sCRP and score on the Mini-Mental State Examination (MMSE), a commonly used screening cognitive measure, was investigated in 540 well functioning, healthy, and cognitively normal elders (age 73 6 6 years). Sociodemographic

Giovanni Ravaglia; Paola Forti; Fabiola Maioli; Nicoletta Brunetti; Mabel Martelli; Lucia Servadei; Luciana Bastagli; Marisa Bianchin; Erminia Mariani



Serum Ratio of Heart-Type Fatty Acid-Binding Protein to Myoglobin  

Microsoft Academic Search

Background: The concentration of heart-type fatty acid-binding protein (hFABP), a promising novel marker for detection of acute or persistent myocardial damage, is significantly influenced by renal clearance and thus has limitations to its usefulness in patients with renal dysfunction. We evaluated whether the serum ratio of hFABP to myoglobin (F\\/M) might be a useful marker for assessing cardiac damage in

Masato Furuhashi; Nobuyuki Ura; Koichi Hasegawa; Hideaki Yoshida; Kazufumi Tsuchihashi; Tomoaki Nakata; Kazuaki Shimamoto



Origin of Fluorescence Lifetimes in Human Serum Albumin. Studies on Native and Denatured Protein  

Microsoft Academic Search

Human serum albumin consists of a single polypeptide of 585 amino acid residues with 1 Trp residue. In the present work, we\\u000a measured fluorescence lifetimes of the protein in both native and denatured states. The results indicate that Trp emission\\u000a occurs with three lifetimes in both states. Lifetimes values and contribution to the global emission decay differ between\\u000a the two

Megdouda Amiri; Kristina Jankeje; Jihad René Albani



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.



Binding of colchicine and thiocolchicoside to human serum proteins and blood cells.  


The binding of 3H-colchicine and its derivative 3H-thiocolchicoside to human serum, purified human proteins and blood cells was studied by equilibrium dialysis and centrifugation. Binding of colchicine and thiocolchicoside to human serum was 38.9 C +/- 4.7 and 12.8 C +/- 5.3%, respectively, essentially to albumin. Protein binding was not dependent on the concentration of either drug between 10(-10) and 10(-5)M. The binding of colchicine and thiocolchicoside to isolated erythrocytes (55 C +/- 5.6 and 16.5 C +/- 2.1%, respectively) decreased markedly in the presence of human serum proteins, i.e. in whole blood (38.7 C +/- 3.1 and 3.4 C +/- 0.8%). Binding of colchicine and thiocolchicoside to other blood cells was very low C < 3%). These binding properties in the blood compartment do not predispose colchicine and thiocolchicoside to be pharmacokinetically sensitive to binding displacement by drug interactions. PMID:7981928

Sabouraud, A; Chappey, O; Dupin, T; Scherrmann, J M



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



Influence of human diets containing casein, soy protein isolate, and soy protein concentrate on serum cholesterol and lipoproteins in humans, rabbits and rats  

Microsoft Academic Search

It is well known that feeding animals such as rabbits with semipurified diets containing animal proteins, as for example casein, results in hypercholesterolemia and atherosclerosis. On the other hand, diets containing vegetable proteins such as soybean protein maintain low levels of serum cholesterol. Little is known about the effects of the type of protein in the diet in humans.This thesis

Raaij van J. M. A



Secretory endometrial protein PP14 in serum from post-menopausal women receiving continuous combined oestradiol-cyproterone acetate: correlation with serum hormone concentrations and bleeding patterns.  


The secretory endometrial protein PP14 was measured in serum from 49 healthy, early post-menopausal women receiving continuous combined oestradiol valerate/cyproterone acetate (2 mg E2V + 1 mg CPA daily) or placebo over a period of 2 years. In the hormone group, serum PP14 increased from 2.1 micrograms/l to a maximum of 8.1 micrograms/l after 1 month of treatment, then fell after 3 months to 3.8 micrograms/l and remained at that level for the rest of the 2-year period. After the first month, the occurrence of uterine bleeding was associated with significantly increased serum PP14 levels. Bleeding was not correlated with the serum concentration of 17 beta-oestradiol (E2) or CPA, or the CPA/E2 ratio. Serum PP14 was significantly dependent on the serum concentration of E2, but not on that of CPA. The present data confirm that serum PP14 levels reflect the secretory phase of the endometrium and that bleeding during continuous combined hormone replacement therapy is probably caused by a sub-optimal hormonal balance. PMID:1388220

Byrjalsen, I; Thormann, L; Riis, B J; Christiansen, C



Determination of protein-ligand binding affinity by NMR: observations from serum albumin model systems.  


The usefulness of bovine serum albumin (BSA) as a model protein for testing NMR methods for the study of protein-ligand interactions is discussed. Isothermal titration calorimetry established the binding affinity and stoichiometry of the specific binding site for L-tryptophan, D-tryptophan, naproxen, ibuprofen, salicylic acid and warfarin. The binding affinities of the same ligands determined by NMR methods are universally weaker (larger KD). This is because the NMR methods are susceptible to interference from additional non-specific binding. The L-tryptophan-BSA and naproxen-BSA systems were the best behaved model systems. PMID:15816062

Fielding, Lee; Rutherford, Samantha; Fletcher, Dan



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



Association of Serum Retinol-Binding Protein 4 and Visceral Adiposity in Chinese Subjects with and without Type 2 Diabetes  

Microsoft Academic Search

Objective: Previous studies have shown that adipose-derived serum retinol-binding protein 4 (RBP4) levels are increased in insulin- resistantmousemodelsandinsubjectswithinsulinresistanceortype 2 diabetes. However, the association of visceral fat and serum RBP4 has not been studied. The purpose of this study was to investigate the relationship between serum RBP4 and regional fat distribution in Chinese subjects with and without type 2 diabetes. Design:

Weiping Jia; Haiya Wu; Yuqian Bao; Chen Wang; Junxi Lu; Jiehua Zhu; Kunsan Xiang


Serum Surfactant Protein-A Is a Strong Predictor of Early Mortality in Idiopathic Pulmonary Fibrosis  

PubMed Central

Background: Serum surfactant protein (SP) A and SP-D had prognostic value for mortality in patients with idiopathic pulmonary fibrosis (IPF) in prior studies before the reclassification of the idiopathic interstitial pneumonias. We hypothesized that baseline serum SP-A and SP-D concentrations would be independently associated with mortality among patients with biopsy-proven IPF and would improve a prediction model for mortality. Methods: We evaluated the association between serum SP-A and SP-D concentrations and mortality in 82 patients with surgical lung biopsy-proven IPF. Regression models with clinical predictors alone and clinical and biomarker predictors were used to predict mortality at 1 year. Results: After controlling for known clinical predictors of mortality, we found that each increase of 49 ng/mL (1 SD) in baseline SP-A level was associated with a 3.3-fold increased risk of mortality (adjusted hazard ratio, 3.27; 95% confidence interval, 1.49 to 7.17; adjusted p = 0.003) in the first year after presentation. We did not observe a statistically significant association between serum SP-D and mortality (adjusted hazard ratio, 2.04; p = 0.053). Regression models demonstrated a significant improvement in the 1-year mortality prediction model when serum SP-A and SP-D (area under the receiving operator curve [AROC], 0.89) were added to the clinical predictors alone (AROC, 0.79; p = 0.03). Conclusions: Increased serum SP-A level is a strong and independent predictor of early mortality among patients with IPF. A prediction model containing SP-A and SP-D was substantially superior to a model with clinical predictors alone.

Kinder, Brent W.; Brown, Kevin K.; McCormack, Francis X.; Ix, Joachim H.; Kervitsky, Alma; Schwarz, Marvin I.; King, Talmadge E.



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

PubMed Central

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

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



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



Protein–Incorporated Serum Total Antioxidant Capacity Measurement by a Modified CUPRAC (CUPric Reducing Antioxidant Capacity) Method  

Microsoft Academic Search

Due to inadequacies in analytical methodology of total antioxidant capacity (TAC) measurement, proteins are initially separated from the human serum matrix by precipitation and are left unmeasured, thereby causing an important “antioxidant gap”. The aim of this work is to measure the TAC of serum with the modified CUPric Reducing Antioxidant Capacity (CUPRAC) method, and to identify the contribution of

Sema Demirci Çekiç; Nilay Kara; Esma Tütem; Kevser Sözgen Ba?kan; Re?at Apak



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

Microsoft Academic Search

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

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



Experiences with dual protein bound aqueous vitamin B12 absorption test in subjects with low serum vitamin B12 concentrations  

Microsoft Academic Search

A dual isotope vitamin B12 absorption test in which vitamin B12 is given both in aqueous solution and bound to protein (chicken serum), was evaluated in 26 controls and 68 patients with subnormal serum vitamin B12 concentrations (19 with pernicious anaemia, 13 with iron deficiency, seven after partial gastrectomy, seven with malabsorptive states, five with folate deficiency, four with chronic

D I Gozzard; D W Dawson; M J Lewis



Concentrations of leptin and C-reactive protein in serum and follicular fluid during assisted reproductive cycles  

Microsoft Academic Search

BACKGROUND: There are only a few studies that have investigated inflammatory processes during ovarian hyperstimulation, with contradictory results especially concerning outcome. The aim of the study was to investigate the inflammatory markers C-reactive protein and leptin in serum and follicular fluid and to correlate these with the outcome. METHODS: One hundred and sixty-two gonadotrophin stimulated cycles were evaluated. Serum concentrations

D. M. Wunder; R. Kretschmer; N. A. Bersinger



Identification of Serum Monocyte Chemoattractant Protein-1 and Prolactin as Potential Tumor Markers in Hepatocellular Carcinoma  

PubMed Central

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.

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



Sirtuin1: A Promising Serum Protein Marker for Early Detection of Alzheimer's Disease  

PubMed Central

Sirtuin (SIRT) pathway has a crucial role in Alzheimer’s disease (AD). The present study evaluated the alterations in serum sirtuin1 (SIRT1) concentration in healthy individuals (young and old) and patients with AD and mild cognitive impairment (MCI). Blood samples were collected from 40 AD and 9 MCI patients as cases and 22 young healthy adults and 22 healthy elderly individuals as controls. Serum SIRT1 was estimated by Surface Plasmon Resonance (SPR), Western Blot and Enzyme Linked Immunosorbent Assay (ELISA). A significant (p<0.0001) decline in SIRT1 concentration was observed in patients with AD (2.27±0.46 ng/µl) and MCI (3.64±0.15 ng/µl) compared to healthy elderly individuals (4.82±0.4 ng/µl). The serum SIRT1 concentration in healthy elderly was also significantly lower (p<0.0001) compared to young healthy controls (8.16±0.87 ng/µl). This study, first of its kind, has demonstrated, decline in serum concentration of SIRT1 in healthy individuals as they age. In patients with AD and MCI the decline was even more pronounced, which provides an opportunity to develop this protein as a predictive marker of AD in early stages with suitable cut off values.

Kumar, Rahul; Chaterjee, Prasun; Sharma, Prakash K.; Singh, Abhay K.; Gupta, Abhishek; Gill, Kamaldeep; Tripathi, Manjari; Dey, Aparajit B.; Dey, Sharmistha




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...


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



Single-Molecule enzyme-linked immunosorbent assay detects serum proteins at subfemtomolar concentrations  

PubMed Central

The detection of single protein molecules1,2 in blood could help identify many new diagnostic protein markers. We report an approach for detecting hundreds to thousands of individual protein molecules simultaneously that enables the detection of very low concentrations of proteins. Proteins are captured on microscopic beads and labeled with an enzyme, such that each bead has either one or zero enzyme-labeled proteins. By isolating these beads in arrays of 50-femtoliter reaction chambers, single proteins can be detected by fluorescence imaging. By singulating molecules in these arrays, ~10–20 enzymes can be detected in 100 ?L (~10?19 M). Single molecule enzyme-linked immunosorbent assays (digital ELISA) based on singulation of enzyme labels enabled the detection of clinically-relevant proteins in serum at concentrations (<10?15 M) much lower than conventional ELISA3-5. Digital ELISA detected prostate specific antigen in all tested sera from patients who had undergone radical prostatectomy, down to 14 fg/mL (0.4 fM).

Rissin, David M.; Kan, Cheuk W.; Campbell, Todd G.; Howes, Stuart C.; Fournier, David R.; Song, Linan; Piech, Tomasz; Patel, Purvish P.; Chang, Lei; Rivnak, Andrew J.; Ferrell, Evan P.; Randall, Jeffrey D.; Provuncher, Gail K.; Walt, David R.; Duffy, David C.



Serum Amino Acids During Gestation of Rhesus Monkeys Fed Two Different Levels of Protein1'3  

Microsoft Academic Search

Female rhesus monkeys were fed a closed formula, puri fied control diet (2.2 g protein per kg body weight) or a low protein diet (0.5 g protein per kg) during pregnancy. Samples of venous blood drawn before conception, on days 50, 100 and 150 after conception, and 7 and 30 days postpartum were analyzed for serum free amino acids. The



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



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



Regulation of inflammation-primed activation of macrophages by two serum factors, vitamin D3-binding protein and albumin.  

PubMed Central

A very small amount (0.0005 to 0.001%) of an ammonium sulfate [50% saturated (NH4)2SO4]-precipitable protein fraction of alpha 2-globulin efficiently supported inflammation-primed activation of macrophages. This fraction contains vitamin D3-binding protein essential for macrophage activation. Comparative macrophage activation studies with fetal calf serum, alpha 2-globulin fraction, 50% (NH4)2SO4 precipitate, and purified bovine vitamin D3-binding protein revealed that fetal calf serum and alpha 2-globulin fraction appear to contain an inhibitor for macrophage activation while ammonium sulfate precipitate contains no inhibitor. This inhibitor was found to be serum albumin. When bovine serum albumin (25 micrograms/ml) was added to a medium supplemented with 0.0005 to 0.05% (NH4)2SO4 precipitate or 1 to 10 ng of vitamin D3-binding protein per ml, activation of macrophages was inhibited.

Yamamoto, N; Kumashiro, R; Yamamoto, M; Willett, N P; Lindsay, D D



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.



Protein signature-based estimation of metagenomic abundances including all domains of life and viruses  

PubMed Central

Motivation: Metagenome analysis requires tools that can estimate the taxonomic abundances in anonymous sequence data over the whole range of biological entities. Because there is usually no prior knowledge about the data composition, not only all domains of life but also viruses have to be included in taxonomic profiling. Such a full-range approach, however, is difficult to realize owing to the limited coverage of available reference data. In particular, archaea and viruses are generally not well represented by current genome databases. Results: We introduce a novel approach to taxonomic profiling of metagenomes that is based on mixture model analysis of protein signatures. Our results on simulated and real data reveal the difficulties of the existing methods when measuring achaeal or viral abundances and show the overall good profiling performance of the protein-based mixture model. As an application example, we provide a large-scale analysis of data from the Human Microbiome Project. This demonstrates the utility of our method as a first instance profiling tool for a fast estimate of the community structure. Availability: Contact: Supplementary information: Supplementary Material is available at Bioinformatics online.

Klingenberg, Heiner; Asshauer, Kathrin Petra; Lingner, Thomas; Meinicke, Peter



Investigation of serum high-sensitive C-reactive protein levels across all mood states in bipolar disorder  

Microsoft Academic Search

There has been an increasing interest in the role of the immune and inflammatory systems in mood disorders. Mood episodes\\u000a are associated with changes in acute phase proteins such as high-sensitivity C-reactive protein (hsCRP). The present study\\u000a investigated serum hsCRP in manic, depressed, and euthymic BD patients as compared to matched healthy controls. Serum hsCRP\\u000a was assessed using an ultrasensitive

Ângelo B. Cunha; Ana C. Andreazza; Fabiano A. Gomes; Benicio N. Frey; Leonardo E. da Silveira; Carlos A. Gonçalves; Flávio Kapczinski



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

Microsoft Academic Search

It has been previously reported that serum levels of 70-kDa heat-shock protein (Hsp70) are elevated in preeclampsia. The aim\\u000a of the present study was to examine whether increased serum Hsp70 levels are related to clinical characteristics and standard\\u000a laboratory parameters of preeclamptic patients, as well as to markers of inflammation (C-reactive protein), endothelial activation\\u000a (von Willebrand factor antigen) or endothelial

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




Microsoft Academic Search

In an acidic medium, the interaction of Fast red VR (FRV) with proteins such as bovine serum albumin (BSA), human serum albumin (HSA), pepsin (Pep), ?-chymotrypsin (Chy), and lysozyme (Lys) was characterized by measuring enhanced resonance light-scattering (RLS) signals. It was found that the enhanced RLS intensities of FRV by proteins at 287.0 nm were in proportion to the concentrations of

Chuan Xiao Yang; Yuan Fang Li; Cheng Zhi Huang



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

PubMed Central

Although biomarkers exist for a range of disease diagnostics, a single low-cost platform exhibiting the required sensitivity, a large dynamic-range and multiplexing capability, and zero sample preparation remains in high demand for a variety of clinical applications. The Interferometric Reflectance Imaging Sensor (IRIS) was utilized to digitally detect and size single gold nanoparticles to identify protein biomarkers in unprocessed serum and blood samples. IRIS is a simple, inexpensive, multiplexed, high-throughput, and label-free optical biosensor that was originally used to quantify biomass captured on a surface with moderate sensitivity. Here we demonstrate detection of ?-lactoglobulin, a cow’s milk whey protein spiked in serum (>10 orders of magnitude) and whole blood (>5 orders of magnitude), at attomolar sensitivity. The clinical utility of IRIS was demonstrated by detecting allergen-specific IgE from microliters of characterized human serum and unprocessed whole blood samples by using secondary antibodies against human IgE labeled with 40 nm gold nanoparticles. To the best of our knowledge, this level of sensitivity over a large dynamic range has not been previously demonstrated. IRIS offers four main advantages compared to existing technologies: it (i) detects proteins from attomolar to nanomolar concentrations in unprocessed biological samples, (ii) unambiguously discriminates nanoparticles tags on a robust and physically large sensor area, (iii) detects protein targets with conjugated very small nanoparticle tags (~40 nm diameter), which minimally affect assay kinetics compared to conventional microparticle tagging methods, and (iv) utilizes components that make the instrument inexpensive, robust, and portable. These features make IRIS an ideal candidate for clinical and diagnostic applications.

Monroe, Margo R.; Daaboul, George G.; Tuysuzoglu, Ahmet; Lopez, Carlos A.; Little, Frederic F.; Unlu, M. Selim



Effects of serum carrier proteins on the growth of pathogenic neisseriae with heme-bound iron.  

PubMed Central

The pathogenic neisseriae can use free heme and hemoglobin as an essential source of iron (Fe) for growth in vitro, but it is unknown whether they can utilize heme bound to human hemopexin or to human serum albumin, or hemoglobin bound to haptoglobin. We found that neither Neisseria meningitidis nor Neisseria gonorrhoeae used bound heme, but bound hemoglobin was used as an Fe source by two meningococcal strains and one gonococcal strain. A second gonococcal strain, previously shown to use free hemoglobin poorly or not at all, also did not grow with hemoglobin-haptoglobin complex as an Fe source. These observations suggest that hemoglobin might act as an Fe source in vivo for many pathogenic neisseriae even when in complexed (bound) form, but heme probably would not support growth in vivo if bound to serum carrier proteins. Images

Dyer, D W; West, E P; Sparling, P F



Serum glial fibrillary acidic protein as a biomarker for intracerebral haemorrhage in patients with acute stroke  

PubMed Central

Background Biomarkers of stroke are an evolving field of clinical research. A serum marker which can differentiate between haemorrhagic and ischaemic stroke in the very early phase would help to optimise acute stroke management. Objective To examine whether serum glial fibrillary acidic protein (GFAP) identifies intracerebral haemorrhage (ICH) in acute stroke patients. Methods A pilot study assessing 135 stroke patients admitted within six hours after symptom onset. Diagnosis of ICH (n?=?42) or ischaemic stroke (n?=?93) was based on brain imaging. GFAP was determined from venous blood samples obtained immediately after admission, using a research immunoassay. Results GFAP was detectable in the serum of 39 patients (34 of 42 (81%) with ICH, and five of 93 (5%) with ischaemic stroke). Serum GFAP was substantially raised in patients with ICH (median 11?ng/l, range 0 to 3096?ng/l) compared with patients with ischaemic stroke (median 0?ng/l, range 0 to 14?ng/l, p<0.001). Using receiver operating characteristic curve analysis , a cut off point of 2.9?ng/l provided a sensitivity of 0.79 and a specificity of 0.98 for the identification of ICH in acute stroke (positive predictive value 0.94, negative predictive value 0.91; p<0.001). Conclusions Serum GFAP can reliably detect ICH in the acute phase of stroke. Further evaluation of the usefulness of GFAP as an early diagnostic marker of ICH is now required, with the aim of optimising cause specific emergency management.

Foerch, C; Curdt, I; Yan, B; Dvorak, F; Hermans, M; Berkefeld, J; Raabe, A; Neumann-Haefelin, T; Steinmetz, H; Sitzer, M



About the structural role of disulfide bridges in serum albumins: evidence from protein simulated unfolding.  


The role of the 17 disulfide (S-S) bridges in preserving the native conformation of human serum albumin (HSA) is investigated by performing classical molecular dynamics (MD) simulations on protein structures with intact and, respectively, reduced S-S bridges. The thermal unfolding simulations predict a clear destabilization of the protein secondary structure upon reduction of the S-S bridges as well as a significant distortion of the tertiary structure that is revealed by the changes in the protein native contacts fraction. The effect of the S-S bridges reduction on the protein compactness was tested by calculating Gibbs free energy profiles with respect to the protein gyration radius. The theoretical results obtained using the OPLS-AA and the AMBER ff03 force fields are in agreement with the available experimental data. Beyond the validation of the simulation method, the results here reported provide new insights into the mechanism of the protein reductive/oxidative unfolding/folding processes. It is predicted that in the native conformation of the protein, the thiol (-SH) groups belonging to the same reduced S-S bridge are located in potential wells that maintain them in contact. The -SH pairs can be dispatched by specific conformational transitions of the peptide chain located in the neighborhood of the cysteine residues. PMID:22899364

Paris, Guillaume; Kraszewski, Sebastian; Ramseyer, Christophe; Enescu, Mironel



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

PubMed Central

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

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



Dietary antioxidant capacity is associated with improved serum antioxidant status and decreased serum C-reactive protein and plasma homocysteine concentrations.  


PURPOSE: To investigate the associations of dietary TAC from diet and supplements with serum antioxidant concentrations and serum C-reactive protein (CRP) and plasma total homocysteine (tHcy) in US adults. METHODS: This was a cross-sectional study. Food consumption data, serum antioxidant levels, and serum CRP and Plasma tHcy concentrations of 4,391 US adults aged ?19 years in the National Health and Nutrition Examination Survey 2001-2002 were analyzed. The USDA flavonoid and proanthocyanidin databases and dietary supplement data as well as antioxidant capacities of 43 antioxidants were also utilized. RESULT: Serum CRP and plasma tHcy concentrations were higher in older adults, smokers, and those with lower non-leisure time physical activity levels (P < 0.05). Energy-adjusted daily total antioxidant capacity (TAC) from diet and supplements was positively associated with serum vitamin E and carotenoid concentrations (P < 0.05). Adjusted odds ratio (OR) for plasma tHcy >13 ?mol/L significantly decreased across quartiles of TAC from diet and supplements (Q1 = 2.18 (1.56-2.77); Q2 = 1.30 (1.00-2.07); Q3 = 1.34 (0.84-2.28); Q4 = 1.00; P for linear trend <0.001). A negative trend across quartiles of TAC from diet and supplements was also observed in OR for serum CRP ?3 mg/L (Q1 = 1.26 (0.97-1.70); Q2 = 1.21 (0.91-1.66); Q3 = 0.97 (0.80-1.24); Q4 = 1.00; P for linear trend <0.05). CONCLUSIONS: These findings indicated that dietary TAC provided an integrated conceptual tool in assessing serum antioxidants and investigating the associations between antioxidant intake and CVD risk. The implicated applicability of dietary TAC needs further validation in prospective cohort studies. PMID:23287847

Yang, Meng; Chung, Sang-Jin; Floegel, Anna; Song, Won O; Koo, Sung I; Chun, Ock K



Hypophysectomy eliminates and growth hormone (GH) maintains the midpregnancy elevation in GH receptor and serum binding protein in the mouse  

SciTech Connect

({sup 125}I)Iodomouse GH (({sup 125}I)iodo-mGH) binding to samples of serum and hepatic microsomal membranes was measured in hypophysectomized pregnant, sham-operated pregnant, intact pregnant, and intact adult virgin mice. Surgeries were carried out on day 11 of pregnancy, and the animals were killed on day 14. The binding of mGH to both serum and hepatic microsomal membranes of intact virgin mice was much lower than to those of intact pregnant mice. In hypophysectomized mice, the mGH-binding capacity of both serum and hepatic microsomes decreased to values similar to those of nonpregnant mice. No significant differences were observed between intact and sham-operated pregnant animals in the maternal serum mGH concentration, the serum GH-binding protein concentration, or the hepatic GH receptor concentration. GH receptor and binding protein-encoding mRNAs were also higher in intact and sham-operated pregnant mice than in virgin and hypophysectomized mice. Hypophysectomized mice were treated with 200 micrograms/day bovine GH, administered by osmotic minipump; after 3 days of treatment, a significant elevation of hepatic GH receptor and serum GH-binding protein levels was observed. These results demonstrate an up-regulation of hepatic GH receptors and serum GH-binding protein by GH during pregnancy in the mouse.

Sanchez-Jimenez, F.; Fielder, P.J.; Martinez, R.R.; Smith, W.C.; Talamantes, F. (Univ. of California, Santa Cruz (USA))



A lack of association between hyperserotonemia and the increased frequency of serum anti-myelin basic protein auto-antibodies in autistic children  

PubMed Central

Background One of the most consistent biological findings in autism is the elevated blood serotonin levels. Immune abnormalities, including autoimmunity with production of brain specific auto-antibodies, are also commonly observed in this disorder. Hyperserotonemia may be one of the contributing factors to autoimmunity in some patients with autism through the reduction of T-helper (Th) 1-type cytokines. We are the first to investigate the possible role of hyperserotonemia in the induction of autoimmunity, as indicated by serum anti-myelin-basic protein (anti-MBP) auto-antibodies, in autism. Methods Serum levels of serotonin and anti-MBP auto-antibodies were measured, by ELISA, in 50 autistic patients, aged between 5 and 12 years, and 30 healthy-matched children. Results Autistic children had significantly higher serum levels of serotonin and anti-MBP auto-antibodies than healthy children (P < 0.001 and P < 0.001, respectively). Increased serum levels of serotonin and anti-MBP auto-antibodies were found in 92% and 80%, respectively of autistic patients. Patients with severe autism had significantly higher serum serotonin levels than children with mild to moderate autism (P < 0.001). Serum serotonin levels had no significant correlations with serum levels of anti-MBP auto-antibodies in autistic patients (P = 0.39). Conclusions Hyperserotonemia may not be one of the contributing factors to the increased frequency of serum anti-MBP auto-antibodies in some autistic children. These data should be treated with caution until further investigations are performed. However, inclusion of serum serotonin levels as a correlate may be useful in other future immune studies in autism to help unravel the long-standing mystery of hyperserotonemia and its possible role in the pathophysiology of this disorder.



Utilization of alkaline phosphatase fusions to identify secreted proteins, including potential efflux proteins and virulence factors from Helicobacter pylori  

Microsoft Academic Search

The targeted genomic strategy of random fusions to a partial gene encoding a signal sequence-deficient fragment of bacterial alkaline phosphatase was utilized to screen for secreted proteins in Helicobacter pylori. The rationale for targeting extracytoplasmic proteins was based on the hypothesis that most virulence factors and vaccine candidates are secreted or exported proteins. In addition, extracytosolic proteins represent good potential

James E Bina; Francis Nano; Robert E. W Hancock



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



Increased Serum Levels of C-Reactive Protein Precede Anastomotic Leakage in Colorectal Surgery  

Microsoft Academic Search

Background  Anastomotic leakage (AL) is a severe complication following colorectal surgery. C-reactive protein (CRP) is considered to\\u000a be an indicator of postoperative complications.\\u000a \\u000a \\u000a \\u000a Materials and methods  Between August 2002 and August 2005 342 colorectal resections with primary anastomosis were performed at the Department of\\u000a General and Vascular Surgery. Johann Wolfgang Goethe-University Frankfurt. For this retrospective study serum CRP was measured\\u000a daily until

Guido WoesteChristine; Christine Müller; Wolf O. Bechstein; Christoph Wullstein



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


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

Bandyopadhyay, A R



Troglitazone reduces hyperglycaemia and selectively acute-phase serum proteins in patients with Type II diabetes  

Microsoft Academic Search

Aims\\/hypothesis. Inflammation could play a part in insulin resistance. Thiazolidinediones, new antidiabetic drugs, possess anti-inflammatory\\u000a effects in vitro. We investigated if acute-phase serum proteins are increased in patients with Type II (non-insulin-dependent)\\u000a diabetes mellitus who had been treated with insulin and whether troglitazone has anti-inflammatory effects in vivo.¶Methods. A total of 27 patients (age 63.0 ± 1.7 years, HbA1 c

P. Ebeling; A.-M. Teppo; H. A. Koistinen; J. Viikari; T. Rönnemaa; M. Nissén; S. Bergkulla; P. Salmela; J. Saltevo; V. A. Koivisto



Enantioselectivity of bovine serum albumin-bonded columns produced with isolated protein fragments  

Microsoft Academic Search

Enantioselectivity of bovine serum albumin (BSA)-bonded columns produced with isolated protein fragments has been investigated. The BSA fragment, BSA-FG75, was isolated by size exclusion chromatography following peptic digest of BSA. The isolated BSA-FG75 was further fractionated to two fractions, BSA-F1 and BSA-F2, by anion-exchange chromatography. BSA-F1 and BSA-F2 had molecular mass of about 35?000 daltons, estimated by matrix-assisted laser desorption

Jun Haginaka; Naoko Kanasugi



Sulfated polysaccharides of brown seaweed Cystoseira canariensis bind to serum myostatin protein.  


Natural sulfated polysaccharides (SPs) derived from brown seaweed comprise a complex group of macromolecules with a wide range of important physiological properties. SPs have been shown to bind and directly regulate the bioactivity of growth factors and cytokines such as basic fibroblast growth factor, interferon, various enzymes and transforming growth factor. Myostatin is a member of the transforming growth factor-beta (TGF-beta) family that acts as a negative regulator of skeletal muscle mass. In this work we demonstrated that SPs isolated from the brown seaweed Cystoseira canariensis bind to the myostatin protein in serum. PMID:14570155

Ramazanov, Zakir; Jimenez del Rio, Miguel; Ziegenfuss, Tim



Identification of serum proteins discriminating colorectal cancer patients and healthy controls using surface-enhanced laser desorption ionisation-time of fl ight mass spectrometry  

Microsoft Academic Search

AIM: To detect the new serum biomarkers for colorectal cancer (CRC) by serum protein profiling with surface- enhanced laser desorption ionisation - time of fl ight mass spectrometry (SELDI-TOF MS). METHODS: Two independent serum sample sets were analysed separately with the ProteinChip technology (set A: 40 CRC + 49 healthy controls; set B: 37 CRC + 31 healthy controls), using

Judith YMN Engwegen; Helgi H Helgason; Annemieke Cats; Nathan Harris; Johannes MG Bonfrer; Jan HM Schellens; Jos H Beijnen


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


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

Pan, Changchuan; Tao, Yalan; Zhao, Ming; Li, Wang; Huang, Zilin; Gao, Jing; Wu, Yanhen; Yu, Jingrui; Wu, Peihong; Xia, Yunfei; Lu, Jin



Serum S100B Protein Levels Are Correlated with Subclinical Neurocognitive Declines after Carotid Endarterectomy  

PubMed Central

OBJECTIVE Carotid endarterectomy (CEA) is an effective means of stroke prevention among appropriately selected patients; however, neuropsychometric testing has revealed subtle cognitive injuries in the early postoperative period. The purpose of this study was to establish whether serum levels of two biochemical markers of cerebral injury were correlated with postoperative declines in neuropsychometric test performance after CEA. METHODS Fifty-five consecutive patients underwent a battery of neuropsychometric tests 24 hours before and 24 hours after elective CEA. Two patients were excluded because of postoperative strokes. The pre- and postoperative serum levels of S100B protein and neuron-specific enolase for injured patients, defined as those who exhibited significant declines in neuropsychometric test performance (n = 12), were compared with the levels for uninjured patients (n = 41). RESULTS There were no significant differences in the baseline S100B levels for the two groups. Injured patients exhibited significantly higher S100B levels, compared with uninjured patients, at 24, 48, and 72 hours after surgery (P < 0.05). There were no significant differences in neuron-specific enolase levels for injured and uninjured patients at any time point. CONCLUSION These data suggest that subtle cerebral injuries after CEA, even in the absence of overt strokes, are associated with significant increases in serum S100B but not neuron-specific enolase levels. Analyses of earlier time points in future studies of subtle cognitive injuries and biochemical markers of cerebral injury after CEA may be revealing.

Connolly, E. Sander; Winfree, Christopher J.; Rampersad, Anita; Sharma, Ruchey; Mack, William J.; Mocco, J; Solomon, Robert A.; Todd, George; Quest, Donald O.; Stern, Yaakov; Heyer, Eric J.



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



Assessment of factors regulating serum growth hormone binding protein in pigs.  


These studies were conducted to examine the influence of several variables on the growth hormone binding protein (GHBP) activity in serum of pigs. Continuous long-term porcine somatotropin (pST) injections (daily for 6 to 7 wk) increased GHBP activity (P less than .05). However, periodic short-term pST injections (daily, every 2nd d, or every 4th d for 2 wk) did not cause a significant change in GHBP levels (P greater than .40). Although fasting seems to reduce liver GH receptors, no difference was observed between fed animals and animals fasted for 5 d (P greater than .30). Between 0 and 6 mo of age, boar and gilt serum GHBP activity were not significantly different from each other but increased with age in both sexes (P less than .0001). There was no significant correlation between serum GHBP and BW at 6 mo of age in this study (P greater than .30). In pregnant sows, GHBP concentrations were highest at the beginning (d 72) of the third trimester (P less than .05). Growth hormone receptor activity reported by other researchers and GHBP activity in this study seem to vary similarly except during fasting, which may indicate alternate regulation of either the GHBP or the GH receptor. PMID:1399889

Mullins, T M; Davis, S L



Serum lipid profile in psoriatic patients: correlation between vascular adhesion protein 1 and lipoprotein (a).  


Psoriasis is a chronic inflammatory skin disease characterized by excessive cellular replication. Apolipoproteins are genetically determined molecule whose role has been implied in cardiovascular pathology. Vascular adhesion protein-1 (VAP-1) is an adhesion molecule with an enzymatic activity that partakes in the migration process of lymphocytes into sites of inflammation. Our purpose was to evaluate the plasma lipid profiles, apolipoproteins (A1, B) and Lp (a) and VAP-1 in order to compare the lipid profile in psoriatic patients with non-affected persons and correlation between VAP-1 and Lp (a). We determined serum concentrations of lipids, lipoproteins , apolipoproteins and VAP-1 in 90 patients with psoriasis and 90 age matched controls. Serum Lp (a), apo A1 and apo B were measured by immunoprecipitation assays, and the lipids and lipoproteins were measured by enzymatic methods.The VAP-1 were measured by ELISA method. The mean levels of total cholesterol, LDL, apo B and VAP-1 in patients with psoriasis were found to be significantly higher than those of healthy subjects (P<0.05. In psoriatic patients, elevation of VAP-1 correlated with elevation of Lp (a) (p = 0.025). This study shows that high serum lipid level and VAP-1, is significantly more common in psoriasis. This fact may be responsible for higher prevalence of cardiovascular accident in psoriatic patients. PMID:22753196

Nemati, Houshang; Khodarahmi, Reza; Rahmani, Ameneh; Ebrahimi, Ali; Amani, Mojtaba; Eftekhari, Kamran



Differential expression profiling of serum proteins and metabolites for biomarker discovery  

NASA Astrophysics Data System (ADS)

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

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



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

PubMed Central

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

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



Nucleocapsid protein N of Lelystad virus: expression by recombinant baculovirus, immunological properties, and suitability for detection of serum antibodies.  

PubMed Central

The ORF7 gene, encoding the nucleocapsid protein N of Lelystad virus (LV), was inserted downstream of the P10 promoter into Autographa californica nuclear polyhedrosis virus (baculovirus). The resulting recombinant baculovirus, designated bac-ORF7, expressed a 15-kDa protein in insect cells. This protein was similar in size to the N protein expressed by LV in CL2621 cells when it was analyzed on sodium dodecyl sulfate-polyacrylamide gels. The N protein expressed by bac-ORF7 was immunoprecipitated with anti-ORF7 was immunoprecipitated with anti-ORF7 peptide serum, porcine convalescent-phase anti-LV serum, and N protein-specific monoclonal antibodies, indicating that this N protein had retained its native antigenic structure. The recombinant N protein was immunogenic in pigs, and the porcine antibodies raised against this protein recognized LV in an immunoperoxidase monolayer assay. However, pigs vaccinated twice with approximately 20 micrograms of N protein were not protected against a challenge with 10(5) 50% tissue culture infective doses of LV. Experimental and field sera directed against various European and North American isolates reacted with the N protein expressed by bac-ORF7 in a blocking enzyme-linked immunosorbent assay. Therefore, the recombinant N protein may be useful for developing diagnostic assays for the detection of serum antibodies directed against different isolates of LV.

Meulenberg, J J; Bende, R J; Pol, J M; Wensvoort, G; Moormann, R J



Is serum retinol binding protein-4: A predictor for diabetes in genetically high risk population?  

PubMed Central

Background: Retinol binding protein-4 (BP-4) a new adipocytokine, specifically binds to retinol, through experimental studies, reported its link between obesity and insulin resistance (IR). But till date no studies are available on influence of genetic predisposition of diabetes on RBP-4 expression. Hence, we aimed to study the influence of genetic predisposition of diabetes on the serum RBP-4 and its role in development of IR and diabetes in genetically high risk population. Materials and Methods: Healthy non diabetic individuals (age 18 to 22) were grouped into Group I: Control (n = 81), whose parents are non diabetic, non hypertensive and does not have any family history of coronary heart diseases. Group II: (n = 157) with one of their parents diabetic and Group III: (n = 47) with both parents diabetic. In all the participants, we estimated fasting serum RBP-4, insulin and glucose. Homeostasis model for assessment-insulin resistance (HOMA-IR) and homeostasis model for assessment-beta cell dysfunction (HOMA-B) were calculated from fasting serum insulin and glucose levels. Results: In this study, we observed significantly higher RBP-4 levels 12.71 ± 2.3 in Group-II and 13.25 ± 2 in Group-III, respectively when compared to Group-I 11.4 ± 1.8 (P < 0.01). RBP-4 showed a significantly strong positive correlation with plasma insulin, glucose and HOMA-IR in genetically high risk population (group II and III) P < 0.01. Linear regression analysis revealed a strong positive association of RBP-4 with parental diabetes even after adjusting for BMI, age and sex (OR 1.53, 95% CI 1.089-1.40). Conclusion: Higher serum RBP-4 and its positive correlation with Insulin, glucose, and HOMA-IR in healthy non diabetic participants of genetically high risk population, indicating its role as predictor for the onset of diabetes in coming future.

Bose, K. Subhash Chandra; Gupta, Shachin K.; Singh, Sandeep



An electrophoretic characterization of serum proteins of the collared peccary (Tayassu tajacu).  


Serum proteins of the collared peccary (Tayassu tajacu) were analyzed by agarose gel electrophoresis for wild adult males and females, nursing young, and reproductively-active females in captivity. Electrophoretic profiles of the adult peccary showed at least six distinct protein bands corresponding to the fractions: albumin, alpha-1, alpha-2, beta-1, beta-2, and gamma globulin. Globulin fractions of the peccary had different mobilities from the domestic swine. The only sexual dimorphisms were associated with the beta globulin:albumin ratio and the albumin:globulin ratio. Ingestion of colostrum in 1-day-old neonates was marked by a very large increase in gamma globulins. The only significant difference between pregnant and lactating females was in the alpha globulin:beta globulin ratio. Lactating females had higher concentrations of alpha-2 globulin than non-pregnant females. PMID:6518761

Lochmiller, R L; Hellgren, E C; Varner, L W; Grant, W E



Feline serum amyloid A protein as an endogenous Toll-like receptor 4 agonist.  


Serum amyloid A (SAA) is one of the major acute phase proteins and a biomarker of infection or inflammation in humans and cats. In humans, cytokine-like functions of SAA protein have been determined, and SAA is considered to be an important factor in immune responses. However, there are no reports about the functions of SAA protein in cats. In the present study, the functions of feline SAA protein on peripheral monocytes were investigated by using TNF-? production as an indicator. In feline peripheral blood monocytes, SAA protein stimulated the transcription of TNF-? within 2h and induced TNF-? secretion in time- and dose-dependent manners. The production of TNF-? by SAA stimulation in feline monocytes was found to be mediated by the activation of nuclear factor-kappa B (NF-?B). Moreover, SAA-stimulated TNF-? production was prevented by a Toll-like receptor 4 (TLR4) antagonist. On the basis of these results, feline SAA was demonstrated to be an endogenous agonist of TLR4 for the stimulation of TNF-? production and secretion by peripheral monocytes. These results suggest that feline SAA can play an important role in the regulation of inflammation and immune responses as it does in humans. PMID:23942262

Tamamoto, Takashi; Ohno, Koichi; Goto-Koshino, Yuko; Tsujimoto, Hajime



Active fragments of the antihemorrhagic protein HSF from serum of habu (Trimeresurus flavoviridis).  


Certain snakes have antihemorrhagic proteins in their sera. Habu serum factor (HSF), an antihemorrhagic protein isolated from the serum of the Japanese habu snake (Trimeresurus flavoviridis) is composed of two cystatin-like domains (D1 and D2) and a His-rich domain, and it inhibits several snake venom metalloproteinases (SVMPs). The activity of HSF can be abolished by trinitrophenylation of Lys residues with 2,4,6-trinitrobenzene sulphonic acid. Upon complex formation of HSF with SVMP, however, the loss of its inhibitory activity by the chemical modification was suppressed, and Lys(15), Lys(41), and Lys(103) residues in HSF were not trinitrophenylated. In order to identify the domain that is critical to the inhibitory activity on SVMPs, native HSF was digested with papain followed by cleavage with cyanogen bromide, yielding a low-molecular mass fragment that was composed of two peptide chains (residues 5-89 and 312-317) linked by a disulfide bond. This fragment inhibited several SVMPs and showed significant antihemorrhagic activity. This indicates that the N-terminal half of D1 is indispensable for the antihemorrhagic activity of HSF. Furthermore, a three-dimensional model of two cystatin-like domains constructed by the homology modeling has indicated that three Lys residues (15, 41, and 103) are exposed to the same surface of HSF molecule. PMID:17222882

Aoki, Narumi; Deshimaru, Masanobu; Terada, Shigeyuki



Two-dimensional acrylamide gel electrophoresis of cancer-patient serum proteins.  


Sera from normal volunteers, patients with a variety of non-neoplastic diseases and patients with malignant or benign tumors were examined by two-dimensional acrylamide gel electrophoresis. In this technique the serum is first separated in an acrylamide gel column followed by a second electrophoresis at right angles to the first separation in a continuous concave 2 to 30 percent gradient acrylamide gel slab. The stained two-dimensional gel slab appears as a "fingerprint" pattern or "map" of the separated serum proteins. Both qualitative and quantitative differences in the fingerprint patterns of cancer-patient sera were observed. The quantitative alterations did not appear specifically associated with malignant tumors. However, several qualitative differences were detected, of which some may represent markers of malignancy as they were not observed in the normal, abnormal and benign tumors control sera. At least one of the abnormal protein stained spots, a prealbumin, appears to be restricted and related in some way to cancer of the lymphoreticular system. These data, although limited, support earlier indications that two-dimensional acrylamide gel electrophoresis offers a new and powerful tool to the cancer scientist for the detection of alterations in cancer-patient sera. A discussion of its possible use to examine other biological fluid and tumor extracts from the cancer patient is presented. PMID:4376659

Wright, G L


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



Analyses of cardiac blood cells and serum proteins with regard to cause of death in forensic autopsy cases  

Microsoft Academic Search

To investigate hematological and serum protein profiles of cadaveric heart blood with regard to the cause of death, serial forensic autopsy cases (n=308, >18 years of age, within 48h postmortem) were examined. Red blood cells (Rbc), hemoglobin (Hb), platelets (Plt), white blood cells (Wbc), total protein (TP) and albumin (Alb) were examined in bilateral cardiac blood. Blood cell counts, collected

Li Quan; Takaki Ishikawa; Tomomi Michiue; Dong-Ri Li; Dong Zhao; Chiemi Yoshida; Jian-Hua Chen; Ayumi Komatsu; Yoko Azuma; Shigeki Sakoda; Bao-Li Zhu; Hitoshi Maeda



Value of measuring serum procalcitonin, C-reactive protein, and mannan antigens to distinguish fungal from bacterial infections  

Microsoft Academic Search

The study presented here was conducted to determine the diagnostic value of measuring procalcitonin, C-reactive protein, and mannan antigens to distinguish fungal from bacterial infections. The sensitivity and specificity of these measurements ranged from 35% to 97%. On days 1 and 3 following the onset of fever, both serum procalcitonin and C-reactive protein levels were lower in patients with fungal

G. L. Petrikkos; S. A. Christofilopoulou; N. K. Tentolouris; E. A. Charvalos; C. J. Kosmidis; G. L. Daikos



Deficiency in the Serum-Derived Hyaluronan-Associated Protein-Hyaluronan Complex Enhances Airway Hyperresponsiveness in a Murine Model of Asthma  

Microsoft Academic Search

Background: Serum-derived hyaluronan (HA)-associated proteins (SHAPs), the heavy chains of inter-?-trypsin inhibitor, covalently bind to HA to form the SHAP-HA complex. The SHAP-HA complex is involved in the pathophysiology of inflammatory diseases, including rheumatoid arthritis. We investigated whether this complex is also involved in airway allergy. Methods: SHAP-HA-deficient (bikunin knockout, KO) mice and wild-type (WT) mice were immunized twice by

Long Zhu; Lisheng Zhuo; Koji Kimata; Etsuro Yamaguchi; Hideto Watanabe; Mark A. Aronica; Vincent C. Hascall; Kenji Baba



Serum interleukin-6, interleukin-8, and ? 2 Microglobulin in early assessment of severity of acute pancreatitis comparison with serum C-reactive protein  

Microsoft Academic Search

The aim of this study was to compare the sensitivity, specificity, and diagnostic accuracy of serum interleukin-6, interleukin-8,?2-microglobulin, and C-reactive protein in the assessment of the severity of acute pancreatitis using commercial kits for their respective assays. Thirty-eight patients with acute pancreatitis (25 men, 13 women, mean age 59 years, range 16–97) were studied; the diagnosis was based on prolonged

Raffaele Pezzilli; Paola Billi; Rita Miniero; Manuela Fiocchi; Onda Cappelletti; Antonio Maria Morselli-Labate; Bahjat Barakat; Giuseppe Sprovieri; Mario Miglioli



Effects of serum protein on ionic exchange between culture medium and microporous hydroxyapatite and silicate-substituted hydroxyapatite.  


It has been proposed that one of the underlying mechanisms contributing to the bioactivity of osteoinductive or osteoconductive calcium phosphates involves the rapid dissolution and net release of calcium and phosphate ions from the matrix as alternatively a precursor to subsequent re-precipitation of a bone-like apatite at the surface and/or to facilitate ion exchange in biochemical processes. In order to confirm and evaluate ion release from sintered hydroxyapatite (HA) and to examine the effect of silicate substitution into the HA lattice on ion exchange under physiological conditions we monitored Ca(2+), PO(4)(3-) and SiO(4)(4-) levels in Earl's minimum essential medium (E-MEM) in the absence (serum-free medium, SFM) or presence (complete medium, C-MEM) of foetal calf serum (FCM), with both microporous HA or 2.6 wt% silicate-substituted HA (SA) sintered discs under both static and semi-dynamic (SD) conditions for up to 28 days. In SFM, variation in Ca(2+) ion concentration was not observed with either disc chemistry or culture conditions. In C-MEM, Ca(2+) ions were released from SA under static and SD conditions whereas with HA Ca(2+) was depleted under SD conditions. PO(4)(3-) depletion occurred in all cases, although it was greater in C-MEM, particularly under SD conditions. SiO(4)(4-) release occurred from SA irrespective of medium or culture conditions but a sustained release only occurred in C-MEM under SD conditions. In conclusion we showed that under physiological conditions the reservoir of exchangeable ions in both HA and SA in the absence of serum proteins is limited, but that the presence of serum proteins facilitated greater ionic exchange, particularly with SA. These observations support the hypothesis that silicate substitution into the HA lattice facilitates a number of ionic interactions between the material and the surrounding physiological environment, including but not limited to silicate ion release, which may play a key role in determining the overall bioactivity and osteoconductivity of the material. However, significant net release of Ca(2+) and PO(4)(3-) was not observed, thus rapid or significant net dissolution of the material is not necessarily a prerequisite for bioactivity in these materials. PMID:21858741

Guth, Katharina; Campion, Charlie; Buckland, Tom; Hing, Karin A



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



Serum C-Reactive Protein Levels in Normal-Weight Polycystic Ovary Syndrome  

PubMed Central

Background/Aims Serum levels of highly sensitive C-reactive protein (hsCRP), a vascular inflammatory marker, may predict the development of cardiovascular disease (CVD) and type 2 diabetes. Women with polycystic ovary syndrome (PCOS) are at greater risk for type 2 diabetes and CVD. The aim of this study was to compare hsCRP levels between normal weight women with PCOS and controls with a normal menstrual cycle and to determine the factors associated with serum hsCRP levels. Methods Thirty-nine lean PCOS patients and 24 healthy, regular cycling women were enrolled in this study. We performed anthropometric measurements, fat computed tomography (CT), and blood sampling to determine blood chemistry and levels of hsCRP, gonadotropins, testosterone, and sex-hormone binding globulin. We also conducted 75-g oral glucose-tolerance test and euglycemic hyperinsulinemic clamp to assess insulin sensitivity. Results Serum hsCRP concentrations were higher in women with PCOS than in women with regular mensturation. However, this difference was no longer significant after adjusting for body mass index (BMI). hsCRP levels were correlated with waist circumference (r=0.46, p<0.01), BMI (r=0.46, p<0.01), visceral fat area (r=0.45, p<0.01), and systolic (r=0.42, p<0.05) and diastolic blood pressure (r=0.39, p<0.05). hsCRP also tended to be negatively associated with insulin-mediated glucose uptake (IMGU) (r=-0.31, p=0.07). A multiple regression analysis revealed that BMI (?=0.29, p<0.05), systolic blood pressure (?=0.39, p<0.01), and IMGU (?=-0.31, p<0.05) predicted serum hsCRP levels in women with PCOS. Conclusions PCOS by itself does not seem to be associated with increased hsCRP levels, whereas known CVD risk factors affect serum hsCRP levels in PCOS.

Oh, Ji Young; Lee, Ji-Ah; Oh, Jee-Young; Sung, Yeon-Ah; Chung, Hyewon



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


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

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



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 1005bp 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 ?124kDa 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



First trimester serum concentrations of placental proteins in singleton and multiple IVF pregnancies: implications for Down syndrome screening  

Microsoft Academic Search

First trimester biochemical trisomy screening is based on serum concentrations of pregnancy-associated plasma protein A (PAPP-A) and human chorionic gonadotrophin (hCG). Our aim was to confirm previously suggested modifications in serum marker concentrations after in vitro fertilisation (IVF) and embryo transfer (ET), and to assess the need of establishing normal medians for trisomy screening in these. We compared 56 singleton

N. A. Bersinger; F. Vanderlick; M. H. Birkhäuser; P. Janecek; D. Wunder



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



Helicobacter pylori Interactions with Host Serum and Extracellular Matrix Proteins: Potential Role in the Infectious Process  

PubMed Central

Helicobacter pylori, a gram-negative spiral-shaped bacterium, specifically colonizes the stomachs of humans. Once established in this harsh ecological niche, it remains there virtually for the entire life of the host. To date, numerous virulence factors responsible for gastric colonization, survival, and tissue damage have been described for this bacterium. Nevertheless, a critical feature of H. pylori is its ability to establish a long-lasting infection. In fact, although good humoral (against many bacterial proteins) and cellular responses are observed, most infected persons are unable to eradicate the infection. A large body of evidence has shown that the interaction between H. pylori and the host is very complex. In addition to the effect of virulence factors on colonization and persistence, binding of specialized bacterial proteins, known as receptins, to certain host molecules (ligands) could explain the success of H. pylori as a chronically persisting pathogen. Some of the reported interactions are of high affinity, as revealed by their calculated dissociation constant. This review examines the binding of host proteins (serum and extracellular matrix proteins) to H. pylori and considers the significance of these interactions in the infectious process. A more thorough understanding of the kinetics of these receptin interactions could provide a new approach to preventing deeper tissue invasion in H. pylori infections and could represent an alternative to antibiotic treatment.

Dubreuil, J. Daniel; Giudice, Giuseppe Del; Rappuoli, Rino



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

PubMed Central

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

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



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


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

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



C-reactive protein (CRP) and serum phospholipase A2 in the assessment of the severity of acute pancreatitis.  

PubMed Central

The present study examines the value of C-reactive protein (CRP) determinations in the assessment of the severity of acute pancreatitis and the correlation of CRP with serum phospholipase A2 activity and the clinical status. Fifty three patients with acute pancreatitis were studied; 17 with haemorrhagic pancreatitis and 36 with a mild form of the disease. S-phospholipase A2 activity increased significantly (p less than 0.05) in patients with fatal pancreatitis but not in those with mild disease. Phospholipase A2 concentrations were below 10 nmol FFA/ml min in mild, while they rose to 20-40 nmol FFA/ml min in haemorrhagic pancreatitis. In fatal cases very high (up to 50-60 nmol FFA/ml min) serum phospholipase A2 concentrations were recorded. The increase in CRP was greater in the patients with severe pancreatitis. One day after admission mean CRP was 280 mg/l in patients with haemorrhagic and 45 mg/l in those with the mild pancreatitis (p less than 0.001). High CRP values also correlated with the prognostic signs indicative of severe pancreatitis. CRP and S-phospholipase A2 determinations are valuable in the early assessment of the severity of acute pancreatitis, but the CRP assay is much easier to include in hospital routine.

Puolakkainen, P; Valtonen, V; Paananen, A; Schroder, T



KEGG and GenomeNet Resources for Predicting Protein Function from Omics Data Including KEGG PLANT Resource  

Microsoft Academic Search

\\u000a With the rise of experimental technologies for omics research in recent years, considerable quantitative data related to transcription,\\u000a protein and metabolism are available for predicting protein functions. To predict protein functions from large omics data,\\u000a reference knowledge databases and bioinformatics tools play considerable roles. KEGG (http:\\/\\/\\/kegg\\/) database we have been establishing is an integrated database of biological systems including genomic,

Toshiaki Tokimatsu; Masaaki Kotera; Susumu Goto; Minoru Kanehisa


Effect of voluntary exercise and dietary protein levels on serum lipoprotein distributions and lecithin: cholesterol acyltransferase (LCAT) activity of mice.  


The effect of voluntary exercise on serum lipoprotein distributions, lecitin:cholesterol acryltransferase (LCAT) activity and serum electrophoretic patterns of mice fed different levels of dietary protein were investigated. Serum cholesterol of all exercise groups (E) showed a lower value than that of the non-exercise groups (NE). Ratios of cholesteryl ester to serum total cholesterol tended to be higher in the exercise groups than in non-exercise groups. High density lipoprotein(HDL)-cholesterol/serum total cholesterol ratios and HDL-cholesterol/low density lipoprotein(LDL)-cholesterol ratios were increased by voluntary exercise. With regard to low density lipoprotein cholesterol levels, there were significant differences between 20% E and 20% NE groups, 4% NE groups, respectively. It was found that HDL fractions in serum lipoprotein patterns of exercise groups differed from those of non-exercise groups. This seemed to be prominent in low protein diet groups. LCAT acitivity showed decreasing values as dietary protein levels decreased and its activity was raised by voluntary exercise in all groups. PMID:7411252

Yashiro, M; Kimura, S



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

Microsoft Academic Search

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

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



Albumin and labile-protein serum concentrations during very-low-calorie diets with different compositions1'2  

Microsoft Academic Search

Circulating concentrations of albumin and the labile proteins prealbumin (PA) and retinol-binding pro- tein (RBP) were evaluated over 20 d in five groups of obese patients. The patients were given four types of very-low-calorie diets (VLCDs) (< 500 kcal\\/d) that provided different amounts of protein or carbohydrate (CHO) plus protein and a conven- tional l200-kcal\\/d hypocaloric diet. Serum albumin concen-

Luca Scalfi; Alessandra Laviano; LouiseA Reed; Renato Borrelli; Franco Contaldo


Use of serum retinol-binding protein for prediction of vitamin A deficiency: effects of HIV1 infection, protein malnutrition, and the acute phase response1-3  

Microsoft Academic Search

Background: Serum retinol is the most commonly used indicator of vitamin A status. Retinol is transported in a 1-to-1 complex with retinol-binding protein (RBP). RBP is easy and inexpensive to measure, and studies have shown a high correlation between concentrations of RBP and concentrations of retinol. The perfor- mance of RBP in the context of infection or protein malnutrition, however,

Jared M Baeten; Barbra A Richardson; Daniel D Bankson; Mark H Wener; Joan K Kreiss; Ludo Lavreys; Kishorchandra Mandaliya; Job J Bwayo; R Scott McClelland


Large-scale delineation of secreted protein biomarkers overexpressed in cancer tissue and serum.  


Genetic alterations in tumor cells often lead to the emergence of growth-stimulatory autocrine and paracrine signals, involving overexpression of secreted peptide growth factors, cytokines, and hormones. Increased levels of these soluble proteins may be exploited for cancer diagnosis and management or as points of therapeutic intervention. Here, we combined the use of controlled vocabulary terms and sequence-based algorithms to predict genes encoding secreted proteins from among approximately 12,500 sequences represented on oligonucleotide microarrays. Expression of these genes was queried in 150 carcinomas from 10 anatomic sites of origin and compared with 46 normal tissues derived from the corresponding sites of tumor origin and other body tissues and organs. Of 74 different genes identified as overexpressed in cancer tissues, several encode proteins with demonstrated clinical diagnostic application, such as alpha-fetoprotein in liver carcinoma, and kallikreins 6 and 10 in ovarian cancer, or therapeutic utility, such as gastrin-releasing peptide/bombesin in lung carcinomas. We show that several of the other candidate genes encode proteins with high levels of tumor-associated expression by immunohistochemistry on tissue microarrays and further demonstrate significantly elevated levels of another novel candidate protein, macrophage inhibitory cytokine 1, a distant member of the transforming growth factor-beta superfamily, in the serum of patients with metastatic prostate, breast, and colorectal carcinomas. Our results suggest that the combination of annotation/protein sequence analysis, transcript profiling, immunohistochemistry, and immunoassay is a powerful approach for delineating candidate biomarkers with potential clinical significance and may be broadly applicable to other human diseases. PMID:12624183

Welsh, John B; Sapinoso, Lisa M; Kern, Suzanne G; Brown, David A; Liu, Tao; Bauskin, Asne R; Ward, Robyn L; Hawkins, Nicholas J; Quinn, David I; Russell, Pamela J; Sutherland, Robert L; Breit, Samuel N; Moskaluk, Christopher A; Frierson, Henry F; Hampton, Garret M



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



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.



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



A Novel Statistical Prognostic Score Model That Includes Serum CXCL5 Levels and Clinical Classification Predicts Risk of Disease Progression and Survival of Nasopharyngeal Carcinoma Patients  

PubMed Central

Background Aberrant expression of C-X-C motif chemokine 5 (CXCL5) contributes to the progression of various cancers. This study analyzed the clinical significance of serum CXCL5 (sCXCL5) levels of nasopharyngeal carcinoma (NPC) patients, with the goal of building a novel prognostic score model. Experimental Design Serum samples were collected prior to treatment from 290 NPC patients for the detection of sCXCL5 with ELISA. Half of the patients (n?=?145) were randomly assigned to the training set to generate the sCXCL5 cutoff point using receiver operator characteristic (ROC) analysis, while the other half (n?=?145) were assigned to the testing set for validation. Associations between sCXCL5 levels and clinical characteristics were analyzed. A prognostic score model was built using independent predictors derived from multivariate analysis. A concordance index (C-Index) was used to evaluate prognostic ability. Results The sCXCL5 cutoff point was 0.805 ng/ml. Sex, age, histology, T classification, clinical classification and local recurrence were not associated with sCXCL5 levels. However, sCXCL5 levels were positively associated with N classification, distant metastasis and disease progression (P<0.05). A high sCXCL5 level predicted poor 6-year overall survival (OS), poor 6-year distant metastasis-free survival (DMFS), and poor 6-year progression-free survival (PFS). A prognostic score model was subsequently constructed based on sCXCL5 levels and clinical classification (C-C model), which are independent predictors of OS, DMFS, and PFS, as confirmed by the multivariate analysis. Furthermore, this novel model successfully divided the patients into four risk subgroups in the training set, the testing set and the entire set of patients. The C-Indices were 0.751 and 0.762 for the training set and the testing set, respectively. Conclusions sCXCL5 level was determined to be an independent prognostic factor for NPC patients. The novel statistical C-C model, which includes sCXCL5 levels and clinical classification, could be helpful in predicting the prognosis of NPC patients.

Lu, Xing; Sun, Rui; Wang, Lin; Zheng, Lisheng; Ye, Yanfang; Bao, Yingna; Xiang, Yanqun; Guo, Xiang



The Cell-Shape Protein Mrec Interacts with Extracytoplasmic Proteins Including Cell Wall Assembly Complexes in Caulobacter crescentus  

Microsoft Academic Search

The bacterial actin homolog, MreB, forms helical cables within the cell that are required for maintenance of a rod shape. These helical structures are thought to be involved in the spatial organization of cell wall (peptidoglycan) synthesizing complexes of penicillin-binding proteins (PBPs). Here, we examined the role of the MreC cell shape protein in this process in Caulobacter crescentus. Subcellular

Arun V. Divakaruni; Rachel R. Ogorzalek Loo; Yongming Xie; Joseph A. Loo; James W. Gober; Lucy Shapiro



The cell-shape protein MreC interacts with extracytoplasmic proteins including cell wall assembly complexes in Caulobacter crescentus  

Microsoft Academic Search

The bacterial actin homolog, MreB, forms helical cables within the cell that are required for maintenance of a rod shape. These helical structures are thought to be involved in the spatial organization of cell wall (peptidoglycan) synthesizing complexes of penicillin-binding proteins (PBPs). Here, we examined the role of the MreC cell shape protein in this process in Caulobacter crescentus. Subcellular

Arun V. Divakaruni; Rachel R. Ogorzalek Loo; Yongming Xie; Joseph A. Loo; James W. Gober



Acute phase protein and cytokine levels in serum and saliva: A comparison of detectable levels and correlations in a depressed and healthy adolescent sample.  


Recent research has examined associations between inflammation and mental health, and has increasingly focused on utilising younger samples to characterise the temporal relationship between inflammatory responses and the emergence of other symptoms. These studies have typically used blood to measure inflammation, although rates of detection for many inflammatory markers appear to be low. Saliva is a safe and low-cost alternative, and adult research has shown that levels of some salivary markers correlate well with those in serum. However, no research has examined this association in young people. This study examined 16 inflammatory markers in serum and saliva in 17 depressed adolescents and 18 healthy controls, aged 13-18years. In general, detection rates were higher in saliva compared to in serum. When non-detectable levels were excluded, serum levels of C-reactive protein (CRP) correlated with salivary CRP (r=0.424, p=0.015), and this correlation appeared to only exist for those individuals with high levels of serum CRP (r=0.599, p=0.014). However, when non-detectable levels were included as zero, salivary levels of CRP, interleukin (IL)-2, IL-12p70, and interferon (IFN)-? correlated with their serum counterparts. No significant clinical group differences in any acute phase proteins or cytokines were present. This study suggests that saliva can be used to measure inflammation in studies with adolescent participants, especially CRP, as it appears to correlate with systemic inflammation for those individuals who are expected to have high levels of inflammation. Implications for future directions in research on salivary inflammatory markers are discussed. PMID:23999491

Byrne, Michelle L; O'Brien-Simpson, Neil M; Reynolds, Eric C; Walsh, Katrina A; Laughton, Katrina; Waloszek, Joanna M; Woods, Michael J; Trinder, John; Allen, Nicholas B



ER-associated protein degradation is a common mechanism underpinning numerous monogenic diseases including Robinow syndrome  

Microsoft Academic Search

Correct folding of nascent polypeptide chains within the ER is critical for function, assembly into multi- subunit complexes and trafficking through the exocytic pathway for secretory and cell surface proteins. This process is rather inefficient, and a substantial proportion of nascent polypeptides is rejected by an ER quality control system and targeted for degradation. In some cases, only a minor

Ying Chen; William P. Bellamy; Miguel C. Seabra; Mark C. Field; Bassam R. Ali



Characteristics of retinol accumulation from serum retinol-binding protein by cultured sertoli cells  

SciTech Connect

The uptake of retinol was examined in cultured Sertoli cells when retinol was provided as a complex with the transport protein retinol-binding protein (RBP). Sertoil cells accumulated ({sup 3}H)retinol in a time- and temperature-dependent manner. The change in rate of retinol accumulation occurred when the cells had accumulated approximately 0.53 pmol of retinol/{mu}g of cellular DNA. Extraction and HPLC analysis of the cell-associated radioactivity yielded retinol and retinyl esters, indicating that a significant proportion of the accumulated retinol was esterified. Excess unlabeled retinol-RBP competed with ({sup 3}H)retinol-RBP for ({sup 3}H)retinol delivery to the cells, indicating that RBP delivery of retinol was a saturable and competable process. However, free ({sup 3}H)retinol associated with Sertoli cells in a noncompetable manner. The transport constant for specific retinol accumulation from RBP was 3.0 {mu}M. Neither iodinated nor reductively methylated RBP was accumulated by or tightly bound to Sertoli cells. Competition studies indicated, however, that protein recognition is important in the retinol uptake process. RBP, CRBP, and CRBP(II) competed with ({sup 3}H)retinol-RBP for ({sup 3}H)retinol accumulation, but free retinol, retinol-bovine serum albumin, and retinol-{beta}-lactoglobulin did not. These studies indicated that Sertoli cell uptake of retinol involved recognition of the retinol-RBP complex at the cell surface with subsequent internalization of retinol, but not RBP.

Shingleton, J.L.; Skinner, M.K.; Ong, D.E. (Vanderbilt Univ. School of Medicine, Nashville, TN (USA))



A comparison of blood factor XII autoactivation in buffer, protein cocktail, serum, and plasma solutions.  


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

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



Thiolated DAB dendrimer/ZnSe nanoparticles for C-reactive protein recognition in human serum.  


A nanocomposite obtained by a thiol DAB-dendrimer (generation 5), coated with fluorescent ZnSe quantum dots, was successfully synthesized for the selective recognition of C-reactive protein. The procedure presented was carried out by a novel, cheap and non-toxic bottom up synthesis. The nanocomposite showed an excitation at 180 nm, with two emission bands at 411 and 465 nm, with a full-width at half-maximum of 336 nm. The Stokes shift was influenced by the presence of coating molecules and the intensity was dependent on pH due to the presence of a charge transfer process. The transmission electron microscopy images demonstrated that the spherical nanoparticles obtained displayed a regular shape of 30 nm size. The fluorescence intensity was markedly quenched by the presence of C-reactive protein, with a dynamic Stern-Volmer constant of 0.036 M(-1). The quenching profile shows that about 51% of the ZnSe QDs are located in the external layer of the thiol dendrimer accessible to the quencher. The precision of the method obtained as relative standard deviation was 3.76% (4 mg L(-1), n=3). This water soluble fluorescent nanocomposite showed a set of favorable properties to be used as a sensor for the C-reactive protein in serum samples, at concentrations of risk levels. PMID:22967596

Algarra, M; Campos, B B; Gomes, D; Alonso, B; Casado, C M; Arrebola, M M; Diez de los Rios, M J; Herrera-Gutiérrez, M E; Seller-Pérez, G; Esteves da Silva, J C G



Air filter devices including nonwoven meshes of electrospun recombinant spider silk proteins.  


Based on the natural sequence of Araneus diadematus Fibroin 4 (ADF4), the recombinant spider silk protein eADF4(C16) has been engineered. This highly repetitive protein has a molecular weight of 48kDa and is soluble in different solvents (hexafluoroisopropanol (HFIP), formic acid and aqueous buffers). eADF4(C16) provides a high potential for various technical applications when processed into morphologies such as films, capsules, particles, hydrogels, coatings, fibers and nonwoven meshes. Due to their chemical stability and controlled morphology, the latter can be used to improve filter materials. In this protocol, we present a procedure to enhance the efficiency of different air filter devices, by deposition of nonwoven meshes of electrospun recombinant spider silk proteins. Electrospinning of eADF4(C16) dissolved in HFIP results in smooth fibers. Variation of the protein concentration (5-25% w/v) results in different fiber diameters (80-1,100 nm) and thus pore sizes of the nonwoven mesh. Post-treatment of eADF4(C16) electrospun from HFIP is necessary since the protein displays a predominantly ?-helical secondary structure in freshly spun fibers, and therefore the fibers are water soluble. Subsequent treatment with ethanol vapor induces formation of water resistant, stable ?-sheet structures, preserving the morphology of the silk fibers and meshes. Secondary structure analysis was performed using Fourier transform infrared spectroscopy (FTIR) and subsequent Fourier self-deconvolution (FSD). The primary goal was to improve the filter efficiency of existing filter substrates by adding silk nonwoven layers on top. To evaluate the influence of electrospinning duration and thus nonwoven layer thickness on the filter efficiency, we performed air permeability tests in combination with particle deposition measurements. The experiments were carried out according to standard protocols. PMID:23685883

Lang, Gregor; Jokisch, Stephan; Scheibel, Thomas



Molecular interactions between serum albumin proteins and Keggin type polyoxometalates studied using luminescence spectroscopy.  


The interaction between the plenary Keggin H3PW12O40, lacunary Keggin K7PW11O39 and the Eu(iii)-substituted Keggin K4EuPW11O39 (Eu-Keggin) type polyoxometalates (POMs), and the proteins human and bovine serum albumin (HSA and BSA) was studied using steady state and time-resolved Eu(iii) luminescence and tryptophan (Trp) fluorescence spectroscopy. The excitation spectrum of the Eu-Keggin POM is dominated by a ligand-to-metal charge transfer band at 291 nm. In the absence of proteins, the number of water molecules coordinated in the first coordination sphere of the Eu(iii) center of Eu-Keggin was determined to be 4, indicating that Eu(iii) occurs as a 1?:?1 isomer in solution. In the presence of HSA or BSA, the number of coordinated water molecules decreased to 0 and 1, respectively, suggesting interaction between the Eu-Keggin POM and the protein surface. As a result of this interaction, a five-fold increase of the hypersensitive (5)D0 ? (7)F2 transition in the luminescence intensity was observed for the Eu-Keggin-HSA complex. The association constants were calculated to be 1.5 × 10(2) M(-1) and 2.0 × 10(3) M(-1) for the Eu-Keggin-HSA and Eu-Keggin-BSA complexes, respectively. Tryptophan fluorescence quenching studies were performed and the quenching constants were calculated using a Stern-Volmer analysis. The obtained values of the quenching constants were 6.1 × 10(4) M(-1) and 2.0 × 10(6) M(-1) for the Eu-Keggin-HSA and Eu-Keggin-BSA complexes, respectively. The surface map of both proteins shows that the cavity containing the tryptophan has a positive surface potential, providing a specific binding site at the surface of albumin proteins for the negatively charged POM. PMID:24064593

Goovaerts, Vincent; Stroobants, Karen; Absillis, Gregory; Parac-Vogt, Tatjana N



Docosahexaenoic acid enhances hepatic serum amyloid A expression via protein kinase A-dependent mechanism.  


Serum amyloid A (SAA) reduces fat deposition in adipocytes and hepatoma cells. Human SAA1 mRNA is increased by docosahexaenoic acid (DHA) treatment in human cells. These studies asked whether DHA decreases fat deposition through SAA1 and explored the mechanisms involved. We demonstrated that DHA increased human SAA1 and C/EBPbeta mRNA expression in human hepatoma cells, SK-HEP-1. Utilizing a promoter deletion assay, we found that a CCAAT/enhancer-binding protein beta (C/EBPbeta)-binding site in the SAA1 promoter region between -242 and -102 bp was critical for DHA-mediated SAA1 expression. Mutation of the putative C/EBPbeta-binding site suppressed the DHA-induced SAA1 promoter activity. The addition of the protein kinase A inhibitor H89 negated the DHA-induced increase in C/EBPbeta protein expression. The up-regulation of SAA1 mRNA and protein by DHA was also inhibited by H89. We also demonstrated that DHA increased protein kinase A (PKA) activities. These data suggest that C/EBPbeta is involved in the DHA-regulated increase in SAA1 expression via PKA-dependent mechanisms. Furthermore, the suppressive effect of DHA on triacylglycerol accumulation was abolished by H89 in SK-HEP-1 cells and adipocytes, indicating that DHA also reduces lipid accumulation via PKA. The observation of increased SAA1 expression coupled with reduced fat accumulation mediated by DHA via PKA suggests that SAA1 is involved in DHA-induced triacylglycerol breakdown. These findings provide new insights into the complicated regulatory network in DHA-mediated lipid metabolism and are useful in developing new approaches to reduce body fat deposition and fatty liver. PMID:19755416

Tai, Chen C; Chen, Ching Y; Lee, Hsuan S; Wang, Ya C; Li, Tsai K; Mersamm, Harry J; Ding, Shih T; Wang, Pei H



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



Reduced serum retinol-binding protein levels in patients with de novo acute myeloid leukaemia.  


Retinol-binding protein (RBP) has been used as a nutritional index for children with acute myeloid leukaemia (AML) in previous studies. However, no studies have yet examined RBP levels in AML patients from all age groups. In this study, AML patients presented with lower RBP concentrations than healthy control subjects and patients with benign haematopathies. A negative association was observed between serum RBP level and peripheral white blood cell count in M4 and M5 AML patients. Moreover, patients carrying the FLT3-ITD mutation and young patients had lower RBP levels than those lacking this mutation and elderly patients. In conclusion, these observations suggest that aberrant retinol levels may be associated with AML. PMID:24129091

Su, Long; Gao, Sujun; Tan, Yehui; Yang, Yan; Liu, Xiaoliang; Yu, Ping; Lin, Hai; Li, Wei



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

PubMed Central

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

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



Radiometric ligand binding assay for C-reactive protein. Complexed C-reactive protein is not detectable in acute phase serum.  

PubMed Central

A radiometric ligand binding assay for human C-reactive protein (CRP) was established using pneumococcal C polysaccharide (CPS) coupled to magnetizable cellulose particles as the solid phase ligand. Competition for binding to the solid phase between 125I-CRP and unlabelled CRP permitted detection of 30 micrograms/l of CRP and the precise assay of concentrations up to 3000 micrograms/l. Identical results were obtained when the assay was used to quantitate isolated pure CRP and pure CRP added to normal human serum. However in vitro addition of known ligands for CRP to acute phase serum resulted in lowering of the apparent CRP concentration in this assay and addition of as little as 1 microgram/l of free CPS or 1 mg/l of lecithin was demonstrable in this way. A combination of the ligand binding assay and the standard electroimmunoassay for CRP was therefore used to test acute phase sera for the presence of CRP complexed in vitro. No evidence of complexed CRP was detected among sera containing between 1-319 mg/l of CRP from patients with Hodgkin's disease (10), rheumatoid arthritis (10), Crohn's disease (19) and various microbial infections (11), including six with subacute bacterial endocarditis. Since it is likely that CRP does form complexes with its ligands in the plasma these results suggest that complexed CRP is rapidly cleared from the circulation.

De Beer, F C; Shine, B; Pepys, M B



CD4 T Cell Tolerance to Human C-reactive Protein, an Inducible Serum Protein, Is Mediated by Medullary Thymic Epithelium  

Microsoft Academic Search

Summary Inducible serum proteins whose concentrations oscillate between nontolerogenic and tolero- genic levels pose a particular challenge to the maintenance of self-tolerance. Temporal restric- tions of intrathymic antigen supply should prevent continuous central tolerization of T cells, in analogy to the spatial limitation imposed by tissue-restricted antigen expression. Major acute- phase proteins such as human C-reactive protein (hCRP) are typical

Ludger Klein; Thomas Klein; Ulrich Rüther; Bruno Kyewski


Periodic Signaling Controlled by an Oscillatory Circuit That Includes Protein Kinases ERK2 and PKA  

Microsoft Academic Search

Self-regulating systems often use robust oscillatory circuits. One such system controls the chemotactic signaling mechanism of Dictyostelium, where pulses of adenosine 3',5'-monophosphate (cAMP) are generated with a periodicity of 7 minutes. We have observed spontaneous oscillations in activation of the mitogen-activated protein (MAP) kinase ERK2 that occur in phase with peaks of cAMP, and we show that ERK2 modulates cAMP

Mineko Maeda; Sijie Lu; Gad Shaulsky; Yuji Miyazaki; Hidekazu Kuwayama; Yoshimasa Tanaka; Adam Kuspa; William F. Loomis



Description of the torcetrapib series of cholesteryl ester transfer protein inhibitors, including mechanism of action  

Microsoft Academic Search

We have identified a series of potent cholesteryl ester transfer protein (CETP) inhibitors, one member of which, torcetrapib, is undergoing phase 3 clinical trials. In this report, we demonstrate that these inhibitors bind specifically to CETP with 1:1 stoichiometry and block both neutrallipidandphospholipid(PL)transferactivities.CETP preincubated with inhibitor subsequently bound both cho- lesteryl ester and PL normally; however, binding of trigly- ceride

Ronald W. Clark; Roger B. Ruggeri; David Cunningham; Mark J. Bamberger



Analysis of protein dynamics within the septate junction reveals a highly stable core protein complex that does not include the basolateral polarity protein Discs large  

PubMed Central

Barrier junctions prevent pathogen invasion and restrict paracellular leakage across epithelial sheets. To understand how one barrier junction, the septate junction (SJ), is regulated in vivo, we used fluorescence recovery after photobleaching (FRAP) to examine SJ protein dynamics in Drosophila. Most SJ-associated proteins, including Coracle, Neurexin IV and Nervana 2, displayed similar, extremely immobile kinetics. Loss of any of these components resulted in dramatically increased mobility of all others, suggesting that they form a single, highly interdependent core complex. Immobilization of SJ core components coincided with formation of the morphological SJ but occurred after their known role in maintaining epithelial polarity, suggesting that these functions are independent. In striking contrast to the core components, the tumor suppressor protein Discs large was much more mobile and its loss did not affect mobility of core SJ proteins, suggesting that it is not a member of this complex, even though it colocalizes with the SJ. Similarly, disruption of endocytosis affected localization of SJ core components, but did not affect their mobility. These results indicate that formation of a stable SJ core complex is separable from its proper subcellular localization, and provide new insights into the complex processes that regulate epithelial polarity and assembly of the SJ.

Oshima, Kenzi; Fehon, Richard G.



Analysis of protein dynamics within the septate junction reveals a highly stable core protein complex that does not include the basolateral polarity protein Discs large.  


Barrier junctions prevent pathogen invasion and restrict paracellular leakage across epithelial sheets. To understand how one barrier junction, the septate junction (SJ), is regulated in vivo, we used fluorescence recovery after photobleaching (FRAP) to examine SJ protein dynamics in Drosophila. Most SJ-associated proteins, including Coracle, Neurexin IV and Nervana 2, displayed similar, extremely immobile kinetics. Loss of any of these components resulted in dramatically increased mobility of all others, suggesting that they form a single, highly interdependent core complex. Immobilization of SJ core components coincided with formation of the morphological SJ but occurred after their known role in maintaining epithelial polarity, suggesting that these functions are independent. In striking contrast to the core components, the tumor suppressor protein Discs large was much more mobile and its loss did not affect mobility of core SJ proteins, suggesting that it is not a member of this complex, even though it colocalizes with the SJ. Similarly, disruption of endocytosis affected localization of SJ core components, but did not affect their mobility. These results indicate that formation of a stable SJ core complex is separable from its proper subcellular localization, and provide new insights into the complex processes that regulate epithelial polarity and assembly of the SJ. PMID:21807950

Oshima, Kenzi; Fehon, Richard G



Serum-protein effects on the detection of the ?-blocker propranolol by ion-transfer voltammetry at a micro-ITIES array  

Microsoft Academic Search

In this work, the effect of the serum protein, bovine serum albumin (BSA), on the detection of propranolol in artificial serum by ion-transfer voltammetry at an array of micro-interfaces between two immiscible electrolyte solutions (?ITIES) is presented. Cyclic voltammetry (CV), differential pulse voltammetry (DPV), and differential pulse stripping voltammetry (DPSV) were examined for the detection of low concentrations of propranolol.

Courtney J. Collins; Conor Lyons; Jörg Strutwolf; Damien W. M. Arrigan



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


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. PMID:10336931

Arima, M; Kanoh, T



TEMED-enhanced photoluminescent imaging of human serum proteins by quantum dots after PAGE.  


Polyacrylamide gel electrophoresis (PAGE) has been one of the most powerful and widely used separation techniques for complex biological samples, whose traditional detection methods include organic dye or silver staining. As a simple, convenient, and ultrasensitive detection of proteins for PAGE, a novel enhanced photoluminescent (PL) imaging method was developed. Thioglycolic acid (TGA)-capped CdTe quantum dots (QDs) and the enhanced reagent of tetramethylethylenediamine (TEMED) are introduced, achieving the direct detection of various proteins in native 1-DE, 2-DE, and SDS gels. Here, we describe the general protocol of TEMED-enhanced PL imaging by QDs, including materials, practical procedures, as well as some notes. PMID:22585516

Na, Na; Ouyang, Jin



Presence of common antigens, including major surface protein epitopes, between the cattle (intraerythrocytic) and tick stages of Anaplasma marginale.  


Epitopes of major surface proteins of the intraerythrocytic cattle stage of Anaplasma marginale were demonstrated in the midgut stage of the organism within the infective tick host Dermacentor andersoni. These proteins were common to all A. marginale isolates tested and at all stages of parasitemia. Sera from cattle immunized with the tick midgut stage of A. marginale immunoprecipitated multiple-erythrocyte-stage proteins, as demonstrated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The major proteins recognized (primarily greater than 14 and less than 200 kilodaltons [kDa]) included two major-erythrocyte-stage surface proteins of 36 and 105 kDa molecular size. To confirm the presence of common tick and erythrocyte A. marginale antigens with the immunized cattle sera, we purified the 36-kDa erythrocyte-stage protein by monoclonal immunoaffinity chromatography and developed an enzyme-linked immunosorbent assay based on the purified protein. All sera from cattle immunized with tick-stage A. marginale and cattle infected with various isolates of A. marginale developed antibodies to the 36-kDa protein. The potential immunoprophylactic, diagnostic, and epidemiologic value of the major epitopes common to both the invertebrate and mammalian stages of A. marginale, especially the 36-kDa protein, is discussed. PMID:2415457

Palmer, G H; Kocan, K M; Barron, S J; Hair, J A; Barbet, A F; Davis, W C; McGuire, T C



The Electrophoretical Determination of Serum Protein Fractions in Lycopene Treated Experimental Diabetic Rats.  


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

Yuksek, Veysel; Dede, Semiha; Ceylan, Ebubekir



Comparing the Efficiencies of Hydrazide Labels in the Study of Protein Carbonylation in Human Serum Albumin  

PubMed Central

In this work, we establish a methodology for comparing the efficiencies of different hydrazide labels for detecting protein carbonyls. We have chosen acrolein- modified human serum albumin as a model. This system provides a convenient means of reproducibly generating carbonylated protein. Five hydrazide-based labels were tested. Three carry a biotin affinity tag and the others are simple fatty acid hydrazides. For the biotin-based labels, the yield of the labeling reaction varies considerably and the most commonly used label, biotin hydrazide, gives the lowest yield. The total MS/MS spectrum counts of modified peptides are similar for all of the biotin-based tags indicating that factors beyond the labeling efficiency are important in determining the effectiveness of the label. In addition, there is a large variation in the number of spectra obtained for specific, modified peptides depending on the nature of the labeling group. This variation implies that the relative dectability of a particular modification site is highly dependent on the tagging reagent, and more importantly, titration schemes aimed at identifying the most reactive site based on its threshold concentration will be biased by the choice of tagging reagent. The fatty acid hydrazides are somewhat more effective than the biotin-based hydrazides in generating identifiable MS/MS spectra, but offer no opportunity for enrichment. For the biotin-based tags, avidin affinity chromatography was used with the tryptic digests and each tag led to similar enrichment levels.

Ugur, Zafer; Coffey, Chelsea M.; Gronert, Scott



A human serum albumin-thioredoxin fusion protein prevents experimental contrast-induced nephropathy.  


Contrast-induced nephropathy (CIN), caused by a combination of the direct tubular toxicity of contrast media, a reduction in medullary blood flow, and the generation of reactive oxygen species, is a serious clinical problem. A need exists for effective strategies for its prevention. Thioredoxin-1 (Trx) is a low-molecular-weight endogenous redox-active protein with a short half-life in the blood due to renal excretion. We produced a long-acting form of Trx as a recombinant human albumin-Trx fusion protein (HSA-Trx) and examined its effectiveness in preventing renal injury in a rat model of ioversol-induced CIN. Compared with saline, a mixture of HSA and Trx, or Trx alone, intravenous HSA-Trx pretreatment significantly attenuated elevations in serum creatinine, blood urea nitrogen, and urinary N-acetyl-?-D-glucosaminidase along with the decrease in creatinine clearance. HSA-Trx also caused a substantial reduction in the histological features of renal tubular injuries and in the number of apoptosis-positive tubular cells. Changes in the markers 8-hydroxy deoxyguanosine and malondialdehyde indicated that HSA-Trx significantly suppressed renal oxidative stress. In HK-2 cells, HSA-Trx decreased the level of reactive oxygen species induced by hydrogen peroxide, and subsequently improved cell viability. Thus, our results suggest that due to its long-acting properties, HSA-Trx has the potential to effectively prevent CIN. PMID:23283135

Kodama, Azusa; Watanabe, Hiroshi; Tanaka, Ryota; Tanaka, Hisae; Chuang, Victor T G; Miyamoto, Yohei; Wu, Qiong; Endo, Masayuki; Hamasaki, Keisuke; Ishima, Yu; Fukagawa, Masafumi; Otagiri, Masaki; Maruyama, Toru



Serum Levels of Pancreatitis-Associated Protein in Digestive Diseases with Special Reference to Gastrointestinal Cancers  

Microsoft Academic Search

The serum levels of pancreatitis-associatedprotein (PAP) were measured in 196 patients withdigestive diseases and 15 healthy subjects by anenzyme-linked immunosorbent assay. The serum PAP levelswere significantly elevated in the patients withgastric, colorectal, biliary tract, hepatocellular, orpancreatic cancers compared with the healthy subjects.After curative resection of the tumor, serum PAP levels were significantly decreased. The serumPAP levels were not related to

Yoshiharu Motoo; Yoshitake Satomura; Ikurou Mouri; Hisatsugu Mouri; Koushiro Ohtsubo; Junta Sakai; Tomoharu Fujii; Hiromi Taga; Yasushi Yamaguchi; Hiroyuki Watanabe; Takashi Okai; Norio Sawabu



Serum evaluation of soluble interferon-?\\/? receptor and high-sensitivity C-reactive protein for diagnosis of the patients with gastrointestinal and hepatobiliary-pancreatic cancer  

Microsoft Academic Search

Serum soluble interferon-?\\/? receptor (sIFN-?\\/?R) and high-sensitivity C-reactive protein (hs-CRP) levels were evaluated in the patients with gastrointestinal and hepatobiliary-pancreatic cancer. We compared the sensitivity and specificity of serum sIFN-?\\/?R with that of serum hs-CRP and evaluated the two diagnostic parameters in combination. Serum sIFN-?\\/?R levels were measured in 92 patients and 25 healthy individuals by enzyme-linked immunosorbent assay. The

Kotaro Miyake; Satoru Imura; Masanori Nishioka; Erdenebulgan Batmunkh; Koji Sugimoto; Yasukazu Ohmoto; Mitsuo Shimada



Differential nephrotoxicity of low molecular weight proteins including Bence Jones proteins in the perfused rat nephron in vivo.  

PubMed Central

To investigate the pathogenetic mechanisms of tubule nephrotoxicity of low molecular weight proteins (LMWP), proximal tubules (PT) of rats were perfused in vivo with artificial tubule fluid (ATF) containing one of five LMWPs: three human Bence Jones proteins (BJP), beta-lactoglobulin (BLG), and rabbit myoglobin (MYG). Volume (JV), chloride (JCl) and glucose (JG) fluxes in these perfused PTs were compared with those determined using ATF alone. In separate experiments, perfused nephrons were examined with electron and immunoelectron microscopy. After exposure to BJP1 or BLG, JV, JCl, and JG were less (P less than 0.05) than corresponding control fluxes. Cell damage of these perfused PTs, along with cellular debris in the distal tubules, was prominent. The PT lysosomes often appeared atypical and contained crystals. In contrast, perfusion with BJP2, BJP3, or MYG did not alter JV, JCl, or JG. These findings were corroborated by the normal ultrastructure of these PTs despite immunohistochemical evidence of endocytosis of the BJPs. Isoelectric point, molecular form, and isotype were not factors associated with PT damage. In addition, proteins with pI less than 7.4 precipitated in the distal nephron, forming acellular casts. Thus, certain nephrotoxic LMWPs damaged the PT, while others precipitated in the distal tubule, obstructing the nephron. These two pathogenetic mechanisms may independently be responsible for tubulointerstitial nephropathy of LMWPs in humans. Images

Sanders, P W; Herrera, G A; Chen, A; Booker, B B; Galla, J H



Serum antibody responses of cattle to iron-regulated outer membrane proteins of Pasteurella haemolytica A1  

Microsoft Academic Search

Serum antibody responses to the 70, 77, and 100 kDa iron-regulated outer membrane proteins (IROMPs) of Pasteurella haemolytica A1 were studied in cattle vaccinated with outer membrane protein (OMP) enriched outer membrane fraction, IROMP-enriched outer membrane fraction or live P. haemolytica. Vaccination with an IROMP-enriched outer membrane fraction stimulated antibodies to the 70 kDa IROMP, whereas vaccination with live P.

Anthony W. Confer; Robert D. McCraw; Janet A. Durham; Rebecca J. Morton; Roger J. Panciera



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\\u000a enabling such a capability, we developed the use of rolling-circle amplification (RCA) to measure the relative levels of proteins\\u000a from two serum samples, labeled with biotin and digoxigenin, respectively, that have been captured on antibody microarrays.\\u000a 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



Serum C-reactive protein and immune responses in dogs inoculated with Bordetella bronchiseptica (phase I cells)  

Microsoft Academic Search

Eight Beagle dogs were inoculated intrabronchially with 5×109 live, avirulent cells ofBordetella bronchiseptica L-414 strain (phase I cells) (B. bronchiseptica) to investigate the serum levels of their C-reactive protein, the white blood cell counts, the antibody responses toB. bronchiseptica in the sera and tracheal secretions, and the effects of prednisolone given to four of the dogs on C-reactive protein (CRP),

S. Yamamoto; T. Shida; M. Honda; Y. Ashida; Y. Rikihisa; M. Odakura; S. Hayashi; M. Nomura; Y. Isayama



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

Microsoft Academic Search

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

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



Correlation Between Proinflammatory Serum Markers: High Sensitivity C-Reactive Protein, Interleukin6 with Disability Score in Acute Ischemic Stroke  

Microsoft Academic Search

Stroke being the third leading cause of death and foremost cause of disability, if potential diagnostic utility of blood borne\\u000a protein biomarkers in predicting acute stroke is established, it would be a substantial adjunct to computerized tomography\\u000a and magnetic resonance imaging which have their own limitations. This study was done to correlate serum Interleukin 6, high\\u000a sensitivity C reactive protein

Anuradha Bharosay; Kiran Saxena; Meena Varma; Vivek Vikram Bharosay; Aparna Pandey


Serum C-Reactive Protein and Self-Reported Stroke Findings From the Third National Health and Nutrition Examination Survey  

Microsoft Academic Search

C-reactive protein may predict the risk of coronary heart disease, but its association with stroke has not been well studied. We used data from the Third National Health and Nutrition Examination Survey, conducted from 1988 to 1994, to examine the association between serum C-reactive protein concentrations and self-reported past history of stroke among 8850 US men and women aged $40

Earl S. Ford; Wayne H. Giles


The application of fluorescein labeled serum proteins (FLSP) to the study of vascular permeability in the brain  

Microsoft Academic Search

1.Vascular permeability in normal and edematous brain tissue was studied by application of the fluorescein labeled serum proteins (FLSP) as well as by the use of free fluorescein isothiocyanate (FITC) marker.2.The electrophoretic studies demonstrated that the binding capacity of the FITC to albumin was of such degree that morphological observations made after injection of the free tracer can be considered

Igor Klatzo; Jaime Miquel; Richard Otenasek



A sensitive and specific quantitation method for determination of serum cardiac Myosin binding protein-C by electrochemiluminescence immunoassay.  


Biomarkers are becoming increasingly more important in clinical decision-making, as well as basic science. Diagnosing myocardial infarction (MI) is largely driven by detecting cardiac-specific proteins in patients' serum or plasma as an indicator of myocardial injury. Having recently shown that cardiac myosin binding protein-C (cMyBP-C) is detectable in the serum after MI, we have proposed it as a potential biomarker for MI. Biomarkers are typically detected by traditional sandwich enzyme-linked immunosorbent assays. However, this technique requires a large sample volume, has a small dynamic range, and can measure only one protein at a time. Here we show a multiplex immunoassay in which three cardiac proteins can be measured simultaneously with high sensitivity. Measuring cMyBP-C in uniplex or together with creatine kinase MB and cardiac troponin I showed comparable sensitivity. This technique uses the Meso Scale Discovery (MSD) method of multiplexing in a 96-well plate combined with electrochemiluminescence for detection. While only small sample volumes are required, high sensitivity and a large dynamic range are achieved. Using this technique, we measured cMyBP-C, creatine kinase MB, and cardiac troponin I levels in serum samples from 16 subjects with MI and compared the results with 16 control subjects. We were able to detect all three markers in these samples and found all three biomarkers to be increased after MI. This technique is, therefore, suitable for the sensitive detection of cardiac biomarkers in serum samples. PMID:23963065

Kuster, Diederik W D; Barefield, David; Govindan, Suresh; Sadayappan, Sakthivel



Serum high-sensitivity C-reactive protein and heat shock protein 27 antibody titers in patients with stroke and 6-month prognosis.  


Serum heat shock protein 27 immunoglobulin G (IgG) antibody titers (anti-HSP27) and high-sensitivity C-reactive protein (hsCRP) concentrations were measured in samples from 168 patients in the first 24 hours after the onset of stroke and 80 age-and sex-matched control participants. In patients with stroke, median serum anti-HSP27 titer was significantly higher than that of the control group (0.18 [0.14-0.28] vs 0.08 [0.04-0.12], P < .001). Median serum hsCRP concentration was also significantly higher in patients compared with the control group (11.43 [8.07-13.53] vs 3.23 [1.66-6.24], P < .001). Serum anti-HSP27 and hsCRP concentrations did not differ significantly among patients with different stroke types. Neither serum anti-HSP27 nor hsCRP levels predicted 6-month prognosis in the patients with stroke. We conclude that serum anti-HSP27 titers and hsCRP concentrations are elevated in patients with stroke but do not distinguish between stroke types or predict 6-month prognosis. PMID:20150172

Azarpazhooh, Mahmoud Reza; Mobarra, Naser; Parizadeh, Syyed Mohammad Reza; Tavallaie, Shima; Bagheri, Mahsa; Rahsepar, Amir Ali; Ghayour-Mobarhan, Majid; Sahebkar, Amirhossein; Ferns, Gordon A A



Preoperative serum C- reactive protein: a prognostic marker in patients with upper urinary tract urothelial carcinoma  

PubMed Central

Background To analyse the prognostic significance of preoperative C-reactive protein (CRP) serum level in patients with upper urinary tract urothelial carcinoma (UUT-UC). Methods We evaluated 158 UUT-UC patients who had undergone surgery in the University Hospital of Hannover (MHH). 143 (89.4%) suffered from cancer in the renal pelvis, 13 (8.1%) patients presented with tumour located in the ureter. A preoperative CRP value was available for 115 patients. The mean (median) follow-up for these patients was 28.3 (15.1) months. Results The median (mean) CRP value of all evaluable patients was 10.0 (40.7) mg/l. The CRP-level, stratified into two subgroups (CRP ?5 vs. >5 mg/l), correlated significantly with muscle invasive tumour stage (36.4 vs. 78.9%; p<0.001), the risk of presenting nodal disease (4.5 vs. 26.8%; p=0.002) and distant metastasis (2.3 vs. 16.9%; p<0.016). The Kaplan-Meier 5-year cancer specific survival (CSS) rates were 54.2 and 26.4% for patients with preoperative CRP levels ? and >5 mg/l, respectively (p<0.006). Next to age and the presence of metastasis, multivariate analysis also identified CRP as a continuous variable as an independent prognosticator for CSS. Conclusions A high preoperative serum CRP level is associated with locally advanced and metastatic disease in patients with UUT-UC. Its routine use could allow better risk stratification and risk-adjusted follow-up of UUT-UC patients.



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

PubMed Central

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

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



Experiences with dual protein bound aqueous vitamin B12 absorption test in subjects with low serum vitamin B12 concentrations.  

PubMed Central

A dual isotope vitamin B12 absorption test in which vitamin B12 is given both in aqueous solution and bound to protein (chicken serum), was evaluated in 26 controls and 68 patients with subnormal serum vitamin B12 concentrations (19 with pernicious anaemia, 13 with iron deficiency, seven after partial gastrectomy, seven with malabsorptive states, five with folate deficiency, four with chronic alcoholism and 13 in whom no cause was apparent). In control patients protein bound absorption decreased with age; isotope excretion was 1.0% or over in those aged under 60 and 0.5% or over in those aged 60 and above. Malabsorption of protein bound vitamin B12 with normal aqueous absorption occurred in five patients with iron deficiency, three with alcoholism, two after partial gastrectomy, two with folate deficiency and in one with a malabsorptive state. In alcoholics abstinence produced an improvement in protein bound absorption. All patients in the group for whom no cause could be found for the subnormal serum vitamin B12 concentration had normal aqueous absorption but four had malabsorption of protein bound vitamin. Although the dual isotope test gave reproducible results and was consistent with the standard Schilling test some anomalies were detected; nine patients had reduced aqueous absorption with normal protein bound absorption. Despite this the dual test may prove useful in determining the importance of a subnormal vitamin B12 concentration where the cause is not clinically apparent. Further development is needed before it can be considered for routine use.

Gozzard, D I; Dawson, D W; Lewis, M J



Experiences with dual protein bound aqueous vitamin B12 absorption test in subjects with low serum vitamin B12 concentrations.  


A dual isotope vitamin B12 absorption test in which vitamin B12 is given both in aqueous solution and bound to protein (chicken serum), was evaluated in 26 controls and 68 patients with subnormal serum vitamin B12 concentrations (19 with pernicious anaemia, 13 with iron deficiency, seven after partial gastrectomy, seven with malabsorptive states, five with folate deficiency, four with chronic alcoholism and 13 in whom no cause was apparent). In control patients protein bound absorption decreased with age; isotope excretion was 1.0% or over in those aged under 60 and 0.5% or over in those aged 60 and above. Malabsorption of protein bound vitamin B12 with normal aqueous absorption occurred in five patients with iron deficiency, three with alcoholism, two after partial gastrectomy, two with folate deficiency and in one with a malabsorptive state. In alcoholics abstinence produced an improvement in protein bound absorption. All patients in the group for whom no cause could be found for the subnormal serum vitamin B12 concentration had normal aqueous absorption but four had malabsorption of protein bound vitamin. Although the dual isotope test gave reproducible results and was consistent with the standard Schilling test some anomalies were detected; nine patients had reduced aqueous absorption with normal protein bound absorption. Despite this the dual test may prove useful in determining the importance of a subnormal vitamin B12 concentration where the cause is not clinically apparent. Further development is needed before it can be considered for routine use. PMID:3611394

Gozzard, D I; Dawson, D W; Lewis, M J



Detection of oxidized methionine in selected proteins, cellular extracts and blood serums by novel anti-methionine sulfoxide antibodies.  


Methionine sulfoxide (MetO) is a common posttranslational modification to proteins occurring in vivo.These modifications are prevalent when reactive oxygen species levels are increased. To enable the detection of MetO in pure and extracted proteins from various sources, we have developed novel antibodies that can recognize MetO-proteins. These antibodies are polyclonal antibodies raised against an oxidized methionine-rich zein protein (MetO-DZS18) that are shown to recognize methionine oxidation in pure proteins and mouse and yeast extracts. Furthermore, mouse serum albumin and immunoglobulin (IgG)were shown to accumulate MetO as function of age especially in serums of methionine sulfoxide reductase A knockout mice. Interestingly, high levels of methionine-oxidized IgG in serums of subjects diagnosed with Alzheimer's disease were detected by western blot analysis using these antibodies. It is suggested that anti-MetO-DZS18 antibodies can be applied in the identification of proteins that undergo methionine oxidation under oxidative stress, aging, or disease state conditions. PMID:19388147

Oien, Derek B; Canello, Tamar; Gabizon, Ruth; Gasset, Maria; Lundquist, Brandi L; Burns, Jeff M; Moskovitz, Jackob



Lights and shadows of proteomic technologies for the study of protein species including isoforms, splicing variants and protein post-translational modifications.  


Recent reviews pinpointed the enormous diversity of proteins found in living organisms, especially in higher eukaryotes. Protein diversity is driven through three main processes: first, at deoxyribonucleic acid (DNA) level (i.e. gene polymorphisms), second, at precursor messenger ribonucleic acid (pre-mRNA) or messenger ribonucleic acid (mRNA) level (i.e. alternative splicing, also termed as differential splicing) and, finally, at the protein level (i.e. PTM). Current proteomic technologies allow the identification, characterization and quantitation of up to several thousands of proteins in a single experiment. Nevertheless, the identification and characterization of protein species using these technologies are still hampered. Here, we review the use of the terms "protein species" and "protein isoform." We evidence that the appropriate selection of the database used for searches can impede or facilitate the identification of protein species. We also describe examples where protein identification search engines systematically fail in the attribution of protein species. We briefly review the characterization of protein species using proteomic technologies including gel-based, gel-free, bottom-up and top-down analysis and discuss their limitations. As an example, we discuss the theoretical characterization of the two human choline kinase species, ?-1 and ?-2, sharing the same catalytic activity but generated by alternative splicing on CHKA gene. PMID:21229583

Casado-Vela, Juan; Cebrián, Arancha; Gómez del Pulgar, Maria Teresa; Sánchez-López, Elsa; Vilaseca, Marta; Menchén, Laura; Diema, Claudia; Sellés-Marchart, Susana; Martínez-Esteso, María José; Yubero, Noemí; Bru-Martínez, Roque; Lacal, Juan Carlos; Lacal, Juan Caelos



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


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

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



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

PubMed Central

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

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



Urinary eosinophil protein X and serum eosinophil cationic protein in infants and young children with atopic dermatitis: Correlation with disease activity  

Microsoft Academic Search

Background: Eosinophil cationic protein (ECP) and eosinophil protein X (EPX) or eosinophil-derived neurotoxin (EDN) are released by eosinophil granulocytes in allergic diseases. Serum ECP (s-ECP) levels have been correlated with disease activity in atopic dermatitis (AD) in adults and young patients, and high urinary EPX (u-EPX) levels in asthmatic patients seem to reflect active disease. A relationship between AD severity

Neri Pucci; Enrico Lombardi; Elio Novembre; Silvia Farina; Roberto Bernardini; Elisabetta Rossi; Tania Favilli; Alberto Vierucci



Abnormal serum thyroid hormones concentration with healthy functional gland: a review on the metabolic role of thyroid hormones transporter proteins.  


Laboratory findings can definitely help the patients not to enter into status, where the damage might be happen due to a miss-diagnosis based on clinical assessment alone. The secondary disease accompanied with thyroid patients should also carefully check out due to the interference which some diseases can cause in the amount of serum thyroid hormone, particularly the free thyroxin. The dilemma over thyroid clinical diagnosis occur due to variation on serum thyroid hormone which initiated by other non-thyroidal disorders which can play an important roles in metabolic disorders of thyroid hormone due to the alteration which occur on the serum level of thyroid hormone transporter proteins. The majority of serum thyroid hormones of up to 95-99% are bound to the carrier proteins mainly to Thyroxin-Binding Globulins (TBG), some transthyretin already known as pre-albumin and albumin which are all synthesis in the liver and any modification which alter their production may alter the status of thyroid hormones. It seems TBG, transthyretin and albumin carries 75, 20, 5% of thyroid hormones within blood circulation, respectively. The dilemma facing the thyroid hormones following disruption of thyroid hormone transporter protein synthesis originate from this fact that any alteration of these protein contribute to the alteration of total thyroid and free serum thyroid hormones which are in fact the biologically active form of thyroid hormones. The subsequent of latter implication result in miss-understanding and miss-diagnosis of thyroid function tests, with possible wrongly thyroid clinical care, followed by undesired therapy of otherwise healthy thyroid. PMID:21874823

Azad, Reza Mansourian



The prevalence of elevated serum C-reactive protein levels in inflammatory and noninflammatory thyroid disease.  


C-reactive protein (CRP) levels have not been routinely used to diagnose thyroid disease, although many thyroid conditions involve inflammation. This study was intended to determine whether CRP levels could differentiate between inflammatory and noninflammatory thyroid conditions, especially between type II inflammatory amiodarone-induced thyrotoxicosis (AIT) and type I iodine-induced AIT. Serum high-sensitivity CRP levels were measured in 100 euthyroid controls (7 taking amiodarone) and 353 patients with one of the following thyroid conditions: AIT, subacute thyroiditis, toxic diffuse goiter, nodular goiter, Hashimoto's thyroiditis, shortterm hypothyroidism, or postpartum thyroiditis. No patients with nontoxic multinodular goiter (n = 34), toxic nodular goiter (n = 23), or toxic diffuse goiter, either untreated (n = 49) or euthyroid while taking methimazole (n = 33), had positive CRP levels (>10 mg/L). The occurrence of positive CRP levels among patients with Hashimoto's thyroiditis (n = 35), short-term hypothyroidism (n = 38), and postpartum thyroiditis (n = 70) did not differ significantly from controls. The occurrence of positive CRP values did not differ significantly between patients with type I and type II AIT and controls. Six of 7 patients (86%) with untreated subacute thyroiditis had positive CRP levels (p < 0.00001). These results indicate that there is only a limited role for measurement of CRP levels in the diagnosis of thyroid diseases other than subacute thyroiditis. PMID:12964969

Pearce, Elizabeth N; Bogazzi, Fausto; Martino, Enio; Brogioni, Sandra; Pardini, Enia; Pellegrini, Giovanni; Parkes, Arthur B; Lazarus, John H; Pinchera, Aldo; Braverman, Lewis E



S100B Protein Detection in Serum Is a Significant Prognostic Factor in Metastatic Melanoma  

Microsoft Academic Search

The serum detection of S100B, a new melanoma marker, has shown clinical significance in early studies. The aim of our study of 1,339 serum samples from 412 different melanoma patients and 107 control patients was to prove the prognostic value of serum S100B levels in melanoma patients at different stages of disease and at follow-up (median: 30 months). Using a

Axel Hauschild; Gudrun Engel; Winfried Brenner; Regine Gläser; Heiner Mönig; Eberhard Henze; Enno Christophers



Changes in Serum Levels of Bone Morphogenic Protein 4 and Inflammatory Cytokines after Bariatric Surgery in Severely Obese Korean Patients with Type 2 Diabetes  

PubMed Central

Serum bone morphogenic protein- (BMP-) 4 levels are associated with human adiposity. The aim of this study was to investigate changes in serum levels of BMP-4 and inflammatory cytokines after Roux-en-Y gastric bypass (RYGB). Fifty-seven patients with type 2 diabetes underwent RYGB. Serum levels of BMP-4 and various inflammatory markers, including high-sensitivity C-reactive protein (hsCRP), free fatty acids (FFAs), and plasminogen activator inhibitor- (PAI-) 1, were measured before and 12 months after RYGB. Remission was defined as glycated hemoglobin <6.5% for at least 1 year in the absence of medications. Levels of PAI-1, hsCRP, and FFAs were significantly decreased at 1 year after RYGB. BMP-4 levels were also significantly lower at 1 year after RYGB than at baseline (P = 0.024). Of the 57 patients, 40 (70%) had diabetes remission at 1 year after surgery (remission group). Compared with patients in the nonremission group, patients in the remission group had lower PAI-1 levels and smaller visceral fat areas at baseline. There was a difference in the change in the BMP-4 level according to remission status. Our data demonstrate a significant beneficial effect of bariatric surgery on established cardiovascular risk factors and a reduction in chronic nonspecific inflammation after surgery.

Kim, Mee Kyoung; Jang, Eun-Hee; Hong, Oak-Kee; Chun, Hyun-Ji; Yoo, Soon-Jib; Baek, Ki-Hyun; Kim, Wook; Kim, Eung Kook; Song, Ki-Ho; Kwon, Hyuk-Sang



LINE-1 ORF1 Protein Localizes in Stress Granules with Other RNA-Binding Proteins, Including Components of RNA Interference RNA-Induced Silencing Complex? †  

PubMed Central

LINE-1 retrotransposons constitute one-fifth of human DNA and have helped shape our genome. A full-length L1 encodes a 40-kDa RNA-binding protein (ORF1p) and a 150-kDa protein (ORF2p) with endonuclease and reverse transcriptase activities. ORF1p is distinctive in forming large cytoplasmic foci, which we identified as cytoplasmic stress granules. A phylogenetically conserved central region of the protein is critical for wild-type localization and retrotransposition. Yeast two-hybrid screens revealed several RNA-binding proteins that coimmunoprecipitate with ORF1p and colocalize with ORF1p in foci. Two of these proteins, YB-1 and hnRNPA1, were previously reported in stress granules. We identified additional proteins associated with stress granules, including DNA-binding protein A, 9G8, and plasminogen activator inhibitor RNA-binding protein 1 (PAI-RBP1). PAI-RBP1 is a homolog of VIG, a part of the Drosophila melanogaster RNA-induced silencing complex (RISC). Other RISC components, including Ago2 and FMRP, also colocalize with PAI-RBP1 and ORF1p. We suggest that targeting ORF1p, and possibly the L1 RNP, to stress granules is a mechanism for controlling retrotransposition and its associated genetic and cellular damage.

Goodier, John L.; Zhang, Lili; Vetter, Melissa R.; Kazazian, Haig H.



Regulation of protein synthesis and degradation in L8 myotubes. Effects of serum, insulin and insulin-like growth factors.  

PubMed Central

We have examined the regulation of protein turnover in rat skeletal myotubes from the L8 cell line. We measured protein synthesis by the rates of incorporation of radiolabelled tyrosine into protein in the presence of a flooding dose of non-radioactive tyrosine. We monitored degradation of proteins labelled with radioactive tyrosine by the release of acid-soluble radioactivity into medium containing excess nonradioactive tyrosine. Extracellular tyrosine pools and intracellular tyrosyl-tRNA equilibrate rapidly during measurements of protein synthesis, and very little reutilization of the radiolabelled tyrosine occurs during degradation measurements. Measured rates of protein synthesis and degradation are constant for several hours, and changes in myotube protein content can be accurately predicted by the measured rates of protein synthesis and degradation. Most of the myotube proteins labelled with radioactive tyrosine for 2 days are degraded, with half-lives (t1/2) of approx. 50 h. A small proportion (less than 2.5%) of the radiolabelled proteins are degraded more rapidly (t1/2 less than 10 h), and, at most, a small proportion (less than 15%) are degraded more slowly (t1/2 greater than 50 h). A variety of agents commonly added to primary muscle cell cultures or to myoblast cell lines (18% Medium 199, 1% chick-embryo extract, antibiotics and antifungal agents) had no effect on rates of protein synthesis or degradation. Horse serum, fetal bovine serum and insulin stimulate protein synthesis and inhibit the degradation of long-lived proteins without affecting the degradation of short-lived proteins. Insulin-like growth factors (IGF)-1 and -2 also stimulate protein synthesis and inhibit protein degradation. The stimulation of protein synthesis and the inhibition of protein degradation are of similar magnitude (a maximum of approx. 2-fold) and display similar sensitivities to a particular anabolic agent. Insulin stimulates protein synthesis and inhibits protein degradation only at supraphysiological doses, whereas IGF-1 and -2 are effective at physiological concentrations. These and other findings suggest that IGFs may be important regulators of skeletal muscle growth during the fetal and early neonatal periods.

Gulve, E A; Dice, J F



Automatic identification of highly conserved family regions and relationships in genome wide datasets including remote protein sequences.  


Identifying shared sequence segments along amino acid sequences generally requires a collection of closely related proteins, most often curated manually from the sequence datasets to suit the purpose at hand. Currently developed statistical methods are strained, however, when the collection contains remote sequences with poor alignment to the rest, or sequences containing multiple domains. In this paper, we propose a completely unsupervised and automated method to identify the shared sequence segments observed in a diverse collection of protein sequences including those present in a smaller fraction of the sequences in the collection, using a combination of sequence alignment, residue conservation scoring and graph-theoretical approaches. Since shared sequence fragments often imply conserved functional or structural attributes, the method produces a table of associations between the sequences and the identified conserved regions that can reveal previously unknown protein families as well as new members to existing ones. We evaluated the biological relevance of the method by clustering the proteins in gold standard datasets and assessing the clustering performance in comparison with previous methods from the literature. We have then applied the proposed method to a genome wide dataset of 17793 human proteins and generated a global association map to each of the 4753 identified conserved regions. Investigations on the major conserved regions revealed that they corresponded strongly to annotated structural domains. This suggests that the method can be useful in predicting novel domains on protein sequences. PMID:24069417

Do?an, Tunca; Karaçal?, Bilge



Automatic Identification of Highly Conserved Family Regions and Relationships in Genome Wide Datasets Including Remote Protein Sequences  

PubMed Central

Identifying shared sequence segments along amino acid sequences generally requires a collection of closely related proteins, most often curated manually from the sequence datasets to suit the purpose at hand. Currently developed statistical methods are strained, however, when the collection contains remote sequences with poor alignment to the rest, or sequences containing multiple domains. In this paper, we propose a completely unsupervised and automated method to identify the shared sequence segments observed in a diverse collection of protein sequences including those present in a smaller fraction of the sequences in the collection, using a combination of sequence alignment, residue conservation scoring and graph-theoretical approaches. Since shared sequence fragments often imply conserved functional or structural attributes, the method produces a table of associations between the sequences and the identified conserved regions that can reveal previously unknown protein families as well as new members to existing ones. We evaluated the biological relevance of the method by clustering the proteins in gold standard datasets and assessing the clustering performance in comparison with previous methods from the literature. We have then applied the proposed method to a genome wide dataset of 17793 human proteins and generated a global association map to each of the 4753 identified conserved regions. Investigations on the major conserved regions revealed that they corresponded strongly to annotated structural domains. This suggests that the method can be useful in predicting novel domains on protein sequences.

Dogan, Tunca; Karacal?, Bilge



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

PubMed Central

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



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

PubMed Central

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

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



Computation of binding energies including their enthalpy and entropy components for protein-ligand complexes using support vector machines.  


Computing binding energies of protein-ligand complexes including their enthalpy and entropy terms by means of computational methods is an appealing approach for selecting initial hits and for further optimization in early stages of drug discovery. Despite the importance, computational predictions of thermodynamic components have evaded attention and reasonable solutions. In this study, support vector machines are used for developing scoring functions to compute binding energies and their enthalpy and entropy components of protein-ligand complexes. The binding energies computed from our newly derived scoring functions have better Pearson's correlation coefficients with experimental data than previously reported scoring functions in benchmarks for protein-ligand complexes from the PDBBind database. The protein-ligand complexes with binding energies dominated by enthalpy or entropy term could be qualitatively classified by the newly derived scoring functions with high accuracy. Furthermore, it is found that the inclusion of comprehensive descriptors based on ligand properties in the scoring functions improved the accuracy of classification as well as the prediction of binding energies including their thermodynamic components. The prediction of binding energies including the enthalpy and entropy components using the support vector machine based scoring functions should be of value in the drug discovery process. PMID:24050538

Koppisetty, Chaitanya A K; Frank, Martin; Kemp, Graham J L; Nyholm, Per-Georg



Serum concentration of C-reactive protein is not a good marker of bronchial hyperresponsiveness  

Microsoft Academic Search

Introduction:  Asthmatic inflammation is responsible for vital features of the disease, including bronchial hyperresponsiveness (BHR). At\\u000a present we do not have precise markers for monitoring asthmatic inflammation. C-reactive protein (CRP), a marker of systemic\\u000a inflammation, seemed to be a factor which could also reflect the level of asthmatic inflammation expressed by BHR. Therefore\\u000a the relationship between CRP concentration and BHR was

Bernard Panaszek; Ewa Liebhart; Jerzy Liebhart; Robert Paw?owicz; Andrzej M. Fal



Follistatin-like protein 1: a serum biochemical marker reflecting the severity of joint damage in patients with osteoarthritis  

PubMed Central

Introduction Follistatin-like protein 1 (FSTL1) is a secreted glycoprotein that has been implicated in arthritis pathogenesis in a mouse model. The aim of this study is to detect FSTL1 expression and to further assess its potential utility as a biomarker of joint damage in osteoarthritis (OA) patients. Methods FSTL1 expression was detected by real-time PCR, western blot and immunohistochemistry (IHC) in the synovial tissues (STs) and by IHC in the articular cartilage from OA patients and control trauma patients. The serum and synovial fluid (SF) FSTL1 concentrations were measured by ELISA in OA patients and control individuals. Linear regression analyses were used to assess correlations between the serum FSTL1 levels and the clinical characteristics in OA patients. Results The FSTL1 mRNA and protein levels were substantially elevated in the STs from OA patients compared with those from control trauma patients. The FSTL1 expression was strong in the cytoplasm of the synovial and capillary endothelial cells of the STs, but weak in the chondrocytes of the articular cartilage from OA patients. Furthermore, the serum and SF FSTL1 concentrations were significantly higher in OA patients than in respective control subjects. Interestingly, the serum and SF FSTL1 levels were markedly higher in female OA patients than in males. Importantly, bivariate regression analysis revealed that the serum FSTL1 levels in female OA patients had significant correlations with Kellgren and Lawrence (KL) grade, joint space narrowing (JSN) and the Western Ontario McMaster and Universities Osteoarthritis (WOMAC) stiffness subscale, an inverse correlation with height, and marginal correlations with the total WOMAC score and the WOMAC function subscale. Multivariate regression analysis revealed that the serum FSTL1 levels correlated independently with KL grade in female OA patients. Bivariate analysis also revealed that the serum FSTL1 levels correlated significantly with age and disease duration, and they correlated marginally with high sensitivity C-reactive protein (hs-CRP) and KL grade in male OA patients. Conclusions Increased FSTL1 expression may be a characteristic of OA patients. FSTL1 is a potential serum biomarker that may reflect the severity of joint damage, and further studies are required to evaluate its potential application for monitoring the course of the disease and the efficacy of therapies in OA patients.



Elevated serum S100B protein in first-episode drug-naive Chinese patients with schizophrenia  

PubMed Central

Mounting evidence suggest that astrocytes might be involved in the pathogenesis of schizophrenia. Of particular interest is S100B, a protein produced primarily by astrocytes that plays a critical role in the maintenance of functional neuronal and astroglial activation. Abnormalities in S100B levels have been associated with schizophrenia. In this study, we examined serum S100B protein levels and the relationship between S100B levels and psychopathological symptoms in first-episode, drug-naďve patients with schizophrenia (SCZ). Sixty-four patients with schizophrenia were compared with 66 age- and sex-matched healthy controls (HC). Psychopathology in the SCZ group was assessed by the Positive and Negative Syndrome Scale (PANSS). Serum S100B protein levels were measured by sandwich enzyme-linked immunosorbent assay (ELISA). The results showed that S100B protein-like immunoreactivity was significantly higher in the SCZ group than the HC group. S100B-like immunoreactivity was correlated with age, duration of illness, and PANSS subscale scores for negative and general psychopathology and total scores. In the SCZ group, serum S100B levels in residual-subtypes were significantly higher than in the paranoid- and disorganized-subtypes. Our findings suggest an upregulation of a marker for astrocyte activity, i.e., S100B, in first-episode medication-free patients with schizophrenia, and thus support the involvement of astrocytes in the pathogenesis of schizophrenia.

Tan, Yunlong; Luo, Xingguang; Yang, Fude; Zhang, Wufang; Wang, Zhiren; Zou, Yizhuang; Zhang, Xiangyang; Wang, Fei; Zuo, Lingjun; Zhou, Dongfeng



Long-term effects of calorie or protein restriction on serum IGF-1 and IGFBP-3 concentration in humans  

PubMed Central

Summary Reduced function mutations in the insulin/IGF-I signaling pathway increase maximal lifespan and health span in many species. Calorie restriction (CR) decreases serum IGF-1 concentration by ~40%, protects against cancer and slows aging in rodents. However, the long-term effects of CR with adequate nutrition on circulating IGF-1 levels in humans are unknown. Here we report data from two long-term CR studies (1 and 6 years) showing that severe CR without malnutrition did not change IGF-1 and IGF-1 : IGFBP-3 ratio levels in humans. In contrast, total and free IGF-1 concentrations were significantly lower in moderately protein-restricted individuals. Reducing protein intake from an average of 1.67 g kg ?1 of body weight per day to 0.95 g kg ?1 of body weight per day for 3 weeks in six volunteers practicing CR resulted in a reduction in serum IGF-1 from 194 ng mL ?1 to 152 ng mL ?1 . These findings demonstrate that, unlike in rodents, long-term severe CR does not reduce serum IGF-1 concentration and IGF-1 : IGFBP-3 ratio in humans. In addition, our data provide evidence that protein intake is a key determinant of circulating IGF-1 levels in humans, and suggest that reduced protein intake may become an important component of anticancer and anti-aging dietary interventions.

Fontana, Luigi; Weiss, Edward P.; Villareal, Dennis T.; Klein, Samuel; Holloszy, John O.



The molecular mechanism of mediation of adsorbed serum proteins to endothelial cells adhesion and growth on biomaterials.  


To explore molecular mechanism of mediation of adsorbed proteins to cell adhesion and growth on biomaterials, this study examined endothelial cell adhesion, morphology and viability on bare and titanium nitride (TiN) coated nickel titanium (NiTi) alloys and chitosan film firstly, and then identified the type and amount of serum proteins adsorbed on the three surfaces by proteomic technology. Subsequently, the mediation role of the identified proteins to cell adhesion and growth was investigated with bioinformatics analyses, and further confirmed by a series of cellular and molecular biological experiments. Results showed that the type and amount of adsorbed serum proteins associated with cell adhesion and growth was obviously higher on the alloys than on the chitosan film, and these proteins mediated endothelial cell adhesion and growth on the alloys via four ways. First, proteins such as adiponectin in the adsorbed protein layer bound with cell surface receptors to generate signal transduction, which activated cell surface integrins through increasing intracellular calcium level. Another way, thrombospondin 1 in the adsorbed protein layer promoted TGF-? signaling pathway activation and enhanced integrins expression. The third, RGD sequence containing proteins such as fibronectin 1, vitronectin and thrombospondin 1 in the adsorbed protein layer bound with activated integrins to activate focal adhesion pathway, increased focal adhesion formation and actin cytoskeleton organization and mediated cell adhesion and spreading. In addition, the activated focal adhesion pathway promoted the expression of cell growth related genes and resulted in cell proliferation. The fourth route, coagulation factor II (F2) and fibronectin 1 in the adsorbed protein layer bound with cell surface F2 receptor and integrin, activated regulation of actin cytoskeleton pathway and regulated actin cytoskeleton organization. PMID:23660250

Yang, Dayun; Lü, Xiaoying; Hong, Ying; Xi, Tingfei; Zhang, Deyuan



Soy protein affects serum insulin and hepatic SREBP-1 mRNA and reduces fatty liver in rats.  


The consumption of soy protein was shown to reduce blood lipids in humans and other animal species. Furthermore, it was shown that the ingestion of soy protein maintains normal insulinemia. Thus, the purpose of the present study was to determine whether soy protein affects the synthesis of lipids in the liver through sterol-regulatory element binding protein-1 (SREBP-1) due to modulation of insulin levels. We first conducted a short-term study in which rats were fed a diet containing 18 g/100 g soy protein or casein for 10 d. Rats fed soy protein had significantly lower serum insulin concentrations than rats fed casein, and this response was accompanied by an elevation in hepatic SREBP-1 mRNA that was 53% lower than that in rats fed casein at d 10. The increase in SREBP-1 mRNA occurred 30 min after consumption of the casein mean, and increased steadily for the next 2 h. We then conducted a second study to assess the long-term effect of soy protein consumption for 150 d on hepatic SREBP-1 expression. Long-term consumption of soy protein maintained normal insulin concentrations compared with rats fed casein, which were hyperinsulinemic. Thus, rats fed the soy protein diet had significantly lower expression of SREBP-1 mRNA than rats fed the casein diet. Soy protein intake also reduced the expression of fatty acid synthase (FAS) and malic enzyme, leading to low hepatic lipid depots of triglycerides and cholesterol, whereas rats fed the casein diet developed fatty liver. These data suggest that soy protein regulates SREBP-1 expression by modulating serum insulin concentration, thus preventing the development of fatty liver. PMID:14988441

Ascencio, Claudia; Torres, Nimbe; Isoard-Acosta, Fernando; Gómez-Pérez, Francisco J; Hernández-Pando, Rogelio; Tovar, Armando R



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


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

Selvaraju, Subhashini; El Rassi, Ziad



Impact of anti-cancer drugs and other determinants on serum protein binding of morphine 6-glucuronide  

PubMed Central

Background and the purpose of the study The aim of the present study was to examine factors that may influence the protein binding of morphine 6-glucuronide (M6G), the most active metabolite of morphine. Methods An enzyme-linked immunoabsorbent assay technique was used to measure the M6G concentration in serum of 18 healthy adults, 18 neonatal and 7 children with cancer. Total and free M6G concentrations were measured following equilibrium dialysis for 3 hrs and at physiological pH at 37°C. The influence of vincristine, methotrexate, 6-mercaptopurine, morphine, human albumin, alpha-1-acid glycoprotein, palmitic acid, oleic acid and pH on M6G protein binding was examined. Results M6G was 66.87±0.73 percent free in human serum at physiological pH and temperature. The percentage free (unbound) was increased significantly by vincristine (4.33%) and methotrexate (9.68%), but 6- mercaptopurine and morphine had no significant effect on it. Free percentages of M6G was reduced by decreasing serum albumin concentration but was unaffected by the presence of alpa-1-acid glycoprotein (AAG) or changes in serum pH. Similar results were obtained in human serum albumin (HAS) solutions. Addition of palmitic acid and oleic acid reduced protein binding significantly by 6.3% and 7.4%, respectively. Major conclusion Although M6G in this study was not highly bounded, but because of its high analgesic potency, any change in its free concentration due to concurrent medication or disease caused significant changes in its effects. This dearth of evidence has been implicated in the reluctance of professionals to be cautious in prescribing them to children, particularly in the neonatal period.

Mashayekhi, S.O.; Ghandforoush-Sattari, M.; Buss, D.C.; Routledge, P.A.; Hain, R.DW.



Haptoglobin phenotyping by polyacrylamide gel isoelectric focusing and its application to simultaneous typing of serum proteins  

Microsoft Academic Search

A simple isoelectric focusing method for haptoglobin (HP) typing is described. Serum was pretreated first with C. perfringens neuraminidase (CPN) and then with dithiothreitol (DTT). The treated serum was subjected to polyacrylamide gel isoelectric focusing (PAGIF), and the band patterns were detected by immunoblotting. The method could be successfully applied to HP typing of bloodstains as old as 2 months.

M. Fukuda; Y. Tamaki; T. Kishida



Serum Lipoproteins Promote Efficient Presentation of the Malaria Virulence Protein PfEMP1 at the Erythrocyte Surface?  

PubMed Central

The virulence of the malaria parasite Plasmodium falciparum is related to its ability to express a family of adhesive proteins known as P. falciparum erythrocyte membrane protein 1 (PfEMP1) at the infected red blood cell surface. The mechanism for the transport and delivery of these adhesins to the erythrocyte membrane is only poorly understood. In this work, we have used specific immune reagents in a flow cytometric assay to monitor the effects of serum components on the surface presentation of PfEMP1. We show that efficient presentation of the A4 and VAR2CSA variants of PfEMP1 is dependent on the presence of serum in the bathing medium during parasite maturation. Lipid-loaded albumin supports parasite growth but allows much less efficient presentation of PfEMP1 at the red blood cell surface. Analysis of the serum components reveals that lipoproteins, especially those of the low-density lipoprotein fraction, promote PfEMP1 presentation. Cytoadhesion of infected erythrocytes to the host cell receptors CD36 and ICAM-1 is also decreased in infected erythrocytes cultured in the absence of serum. The defect appears to be in the transfer of PfEMP1 from parasite-derived structures known as the Maurer's clefts to the erythrocyte membrane or in surface conformation rather than a down-regulation or switching of particular PfEMP1 variants.

Frankland, Sarah; Elliott, Salenna R.; Yosaatmadja, Francisca; Beeson, James G.; Rogerson, Stephen J.; Adisa, Akinola; Tilley, Leann



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

NASA Astrophysics Data System (ADS)

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

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



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

Microsoft Academic Search

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

Martine Szyper Kravitz; Milena Pitashny; Yehuda Shoenfeld



Improving the scoring of protein-ligand binding affinity by including the effects of structural water and electronic polarization.  


Docking programs that use scoring functions to estimate binding affinities of small molecules to biological targets are widely applied in drug design and drug screening with partial success. But accurate and efficient scoring functions for protein-ligand binding affinity still present a grand challenge to computational chemists. In this study, the polarized protein-specific charge model (PPC) is incorporated into the molecular mechanics/Poisson-Boltzmann surface area (MM/PBSA) method to rescore the binding poses of some protein-ligand complexes, for which docking programs, such as Autodock, could not predict their binding modes correctly. Different sampling techniques (single minimized conformation and multiple molecular dynamics (MD) snapshots) are used to test the performance of MM/PBSA combined with the PPC model. Our results show the availability and effectiveness of this approach in correctly ranking the binding poses. More importantly, the bridging water molecules are found to play an important role in correctly determining the protein-ligand binding modes. Explicitly including these bridging water molecules in MM/PBSA calculations improves the prediction accuracy significantly. Our study sheds light on the importance of both bridging water molecules and the electronic polarization in the development of more reliable scoring functions for predicting molecular docking and protein-ligand binding affinity. PMID:23651068

Liu, Jinfeng; He, Xiao; Zhang, John Z H



Serum protein and enzyme levels in rats following administration of ethanolic leaf extract of Ageratum conyzoides (goat weed).  


The The potential hepatotoxic effects following oral administration of ethanolic leaf extract of Ageratum conyzoides (goat weed) was investigated in albino Wistar rats. Twenty eight (28) adult male Wistar rats were uniformly divided into four groups of seven rats each. Group 1 served as control while groups 2, 3 and 4 were respectively gavaged with 200 mg/kg body weight, 400 mg/kg body weight and 600 mg/kg body weight of the extract daily for 21 days. At the end of treatments, animals were sacrificed, serum and liver tissues obtained for assay of total protein concentration and levels of ALT, AST and ALP. Results showed that treatment of rats with the respective doses of the extract did not significantly alter the serum and liver levels of total protein, ALT, AST and ALP in all test groups. This result suggests that ingestion of the extract may not be toxic at the doses investigated. PMID:20234750

Antai, A B; Eyong, E U; Eteng, M U; Itam, E H; Eko, M E; Ita, S O



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

PubMed Central

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

Swamy, Narasimha; Ray, Rahul



A new radioimmunoassay to measure rat insulin-like growth factor binding protein-4 (IGFBP-4) in serum, wound fluid and conditioned media.  


Insulin-like growth factor binding protein-4 (IGFBP-4) is, like the other five IGFBPs, a critical regulator of the activity of insulin-like growth factor-I (IGF-I) and IGF-II. Because of the possible role of IGFBP-4 in multiple systems including reproduction, pregnancy, bone formation and wound healing, we developed a radioimmunoassay (RIA) to measure the concentration of IGFBP-4 in the serum, extracellular fluid and conditioned media of cells of rodents so that these animal models could be better exploited. Rat IGFBP-4 was expressed in Escherichia coli and purified to homogeneity. The recombinant material was used to raise a rabbit polyclonal antibody against rat IGFBP-4 and for iodination and standards. The RIA developed was sensitive to less than 1 ng/mL of rat IGFBP-4 and IGF-I did not interfere. There was no cross-reactivity with other rat IGFBPs on immuno-Western analysis of serum and wound fluid. IGFBP-4 from mouse serum did cross-react in our assay; however, serum from horse, pig or human showed no cross-reaction. Human IGFBP-1 and IGFBP-3 showed a very weak cross-reactivity. Serum IGFBP-4 levels showed no gender difference but did reveal a significant 66% increase in older rats. During the course of wound healing, which is IGF-I dependent, IGFBP-4 showed no changes. In conclusion an RIA for rat IGFBP-4 has been developed that specifically measures the concentration of IGFBP-4 in the sera, extracellular fluid and conditioned media of rodents. With this assay, the role of IGFBP-4 and IGFs in growth, development and the function of multiple systems can be further investigated using rodent models. PMID:11437474

Chelius, D; Spencer, E M




PubMed Central

OBJECTIVE Elevated concentrations of Interleukin-6 (IL-6), C-reactive protein (CRP), and Matrix Metalloproteinase-9 (MMP-9) in fetal and neonatal compartments have been associated with an increased risk for preterm birth (PTB) and/or neonatal morbidity. The purpose of this study was to determine if the maternal serum concentration of IL-6, CRP, and MMP-9 in women at risk for preterm birth (PTB) who are not in labor, and have intact membranes, are associated with an increased risk for preterm birth < 32 weeks and/or neonatal morbidity. STUDY DESIGN Maternal serum samples collected from 475 patients enrolled in a multicenter randomized controlled trial of single versus weekly corticosteroids for women at increased risk for preterm delivery were assayed. Serum was collected at randomization (24-32 weeks gestation). Maternal serum concentrations of IL-6, CRP, and MMP-9 were subsequently determined using enzyme-linked immunoassays. Multivariate logistic regression analysis was performed to explore the relationship between maternal serum concentrations of IL-6, CRP and MMP-9, and preterm birth < 32 weeks, Respiratory Distress Syndrome (RDS), Chronic Lung Disease (CLD), Intraventricular Hemorrhage (IVH), Necrotizing Enterocolitis (NEC) and Any Sepsis (S). RESULTS Maternal serum concentrations of IL-6 and CRP, but not MMP-9, above the 90th percentile, at the time of randomization, were associated with preterm birth < 32 weeks. In contrast, there was no significant relationship between RDS and NEC and the maternal serum concentration of IL-6, CRP, or MMP-9 (univariate analysis). The development of CLD was associated with a high (above 90th percentile) IL-6 and CRP in maternal serum, even after adjustment for gestational age (GA) at randomization, and treatment group. However, when GA at delivery was added to the model, this finding was non-significant. Neonatal sepsis was more frequent in neonates born to mothers with a high maternal serum concentration of CRP (above >90th percentile). However, there was no significant association after adjustment for GA at randomization, and treatment group. Logistic regression analysis for each analyte indicated that high maternal serum concentrations of IL-6 and CRP, but not MMP-9, were associated with an increased risk of IVH (O.R. 4.60, 95% C.I. 1.86-10.68; O.R. 4.07, 95% C.I. 1.63-9.50), after adjusting for GA at randomization and treatment group. Most babies (25/30) had grade I IVH. When GA at delivery was included, elevated IL-6 remained significantly associated with IVH (O.R. 2.77, 95% C.I. 1.02-7.09). CONCLUSION An elevated maternal serum concentration of IL-6 and CRP are risk factors for preterm birth < 32 weeks and subsequent development of neonatal IVH. An elevated maternal serum IL-6 appears to confer additional risk for IVH even after adjusting for gestational age at delivery.

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



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

PubMed Central

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

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



Clinical significances of preoperative serum interleukin-6 and C-reactive protein level in operable gastric cancer  

Microsoft Academic Search

BACKGROUND: The interleukin-6 (IL-6) pathway is one of the mechanisms that link inflammation and angiogenesis to malignancy. Because the C-reactive protein (CRP) is a representative marker for inflammation, CRP has recently been associated with the progression of disease in many cancer types. The principal objective of this study was to determine the preoperative serum levels of IL-6 and CRP in

Do-Kyong Kim; Sung Yong Oh; Hyuk-Chan Kwon; Suee Lee; Kyung A Kwon; Byung Geun Kim; Seong-Geun Kim; Sung-Hyun Kim; Jin Seok Jang; Min Chan Kim; Kyeong Hee Kim; Jin-Yeong Han; Hyo-Jin Kim



Serum levels of autoantibodies against monomeric C-reactive protein are correlated with disease activity in systemic lupus erythematosus  

Microsoft Academic Search

This study was performed to investigate the relation between IgG autoantibodies against human C-reactive protein (anti-CRP) and disease activity measures in serial serum samples from 10 patients with systemic lupus erythematosus (SLE), of whom four had active kidney involvement during the study period. The presence of anti-CRP was analysed by enzyme-linked immunosorbent assay. The cut-off for positive anti-CRP test was

Christopher Sjöwall; Anders A Bengtsson; Gunnar Sturfelt; Thomas Skogh



Diet, serum insulin-like growth factor-I and IGF-binding protein-3 in European women  

Microsoft Academic Search

Objective:The aim of this study was to examine the relationship of diet with serum insulin-like growth factor-I (IGF-I) and IGF-binding protein-3 in women.Design:Cross-sectional study.Setting and subjects:The population are 2109 women who were control subjects in a case–control study of breast cancer nested in the European Prospective Investigation into Cancer and Nutrition. Control subjects were randomly chosen among risk sets consisting

T Norat; L Dossus; S Rinaldi; K Overvad; H Grřnbćk; A Tjřnneland; A Olsen; F Clavel-Chapelon; M C Boutron-Ruault; H Boeing; P H Lahmann; J Linseisen; G Nagel; A Trichopoulou; D Trichopoulos; V Kalapothaki; S Sieri; D Palli; S Panico; R Tumino; C Sacerdote; H B Bueno-de-Mesquita; P H M Peeters; C H van Gils; A Agudo; P Amiano; E Ardanoz; C Martinez; R Quirós; M J Tormo; S Bingham; T J Key; N E Allen; P Ferrari; N Slimani; E Riboli; R Kaaks



Effect of alcohol consumption on serum concentration of 25-hydroxyvitamin D3 , retinol, and retinol-binding protein?3  

Microsoft Academic Search

The effect of chronic alcohol consumption on the concentration of 25- hydroxyvitamin D3 , total retinol, and retinol-binding protein in serum was studied in chronic alcoholics (n = 12) and controls (n = 19). Ethanol intake during the last year was 178 ± I 16 and 3.7 ± 4.5 g\\/day, respectively (p < 0.002). Of the alcoholics, 58% had a

Gunn-Elin Aa Bjrneboe; Jon Johnsen; Anders Bjrneboe; Brigitte Rousseau; Jan I Pedersen; Kaare R Norum; Jrg Mrland; Christian A Drevon


Preoperative serum retinol-binding protein 4 is associated with the prognosis of patients with hepatocellular carcinoma after curative resection  

Microsoft Academic Search

Purpose  Metabolic syndrome and insulin resistance have been linked to increased risk of occurrence and mortality of hepatocellular\\u000a carcinoma (HCC). Recently, retinol-binding protein 4 (RBP4) was clarified as a specific serological marker of insulin resistance.\\u000a The aim of this study was to determine whether serum RBP4 could be used as a potential marker for predicting prognosis in\\u000a patients with HCC after

Dan-Dan Wang; Yi-Ming Zhao; Lu Wang; Guang Ren; Fei Wang; Zu-Guang Xia; Xi-Long Wang; Tao Zhang; Qi Pan; Zhi Dai; Ju-Ping Chen; Hai-Yan Dai; Wei Zhang; Hong-Wei He; Jia-Min Zhou; Guang-Yu Tang; Jian Zhou; Jia Fan; Zhao-You Tang



Rhizopus oligosporus grown on natural rubber waste serum for production of single cell protein: a preliminary study  

Microsoft Academic Search

Maximum production of mycelium and utilization of total organic carbon byR. oligosporus grown on natural rubber waste serum was achieved at 28°C with an inoculum size of 7.5% (v\\/v) and grown for 144 h with an initial pH of 4.0. The maximum production of total crude protein, however, was when culture medium was inoculated with 2% (v\\/v) of spore suspension

M. S. Mahat; I. C. MacRae



Binding characteristics of bovine serum albumin encapsulated in sol-gel glasses: An alternative for protein interaction studies  

Microsoft Academic Search

Silica glasses doped with 500–700?g of bovine serum albumin were prepared by the sol-gel method; two pH conditions (pH 5 and 7) were assayed for protein encapsulation. Both biomaterials showed a highly porous structure, with pore sizes in the range 5–28nm. Columns packed with the ground biogels were on-line coupled to a C18 HPLC column for evaluation of the entrapped

Luz E. Vera-Avila; Erika García-Salgado; Martha P. García de Llasera; Araceli Peńa-Alvarez



Agouti-Related Protein-Like Immunoreactivity: Characterization of Release from Hypothalamic Tissue and Presence in Serum  

Microsoft Academic Search

A novel RIA was used to examine the release of agouti-related protein-like immunoreactivity (AGRP-LI) from perfused rat hypotha- lamic tissue slices and to characterize AGRP-LI in rat serum. A continuous low level basal AGRP-LI release was observed from hy- pothalami of rats fed ad libitum before the rats were killed. Basal AGRP-LI release was 3-fold greater in rats fasted 48




Preoperative plasma plasminogen activator inhibitor type-1 and serum C-reactive protein levels in patients with colorectal cancer  

Microsoft Academic Search

Background: Preoperative plasma plasminogen activator inhibitor-1 (PAI-1) is a prognostic variable in patients with colorectal cancer.\\u000a It has been suggested, however, that plasma PAI-1 is a nonspecific prognostic parameter similar to the acute-phase reactant\\u000a C-reactive protein (CRP). In the present study we analyzed the association between plasma PAI-1 and serum CRP in patients\\u000a scheduled for elective resection of colorectal cancer.

Hans Jřrgen Nielsen; Ib Jarle Christensen; Steen Sřrensen; Flemming Moesgaard; Nils Brtinner



C-reactive protein (CRP) and serum phospholipase A2 in the assessment of the severity of acute pancreatitis  

Microsoft Academic Search

The present study examines the value of C-reactive protein (CRP) determinations in the assessment of the severity of acute pancreatitis and the correlation of CRP with serum phospholipase A2 activity and the clinical status. Fifty three patients with acute pancreatitis were studied; 17 with haemorrhagic pancreatitis and 36 with a mild form of the disease. S-phospholipase A2 activity increased significantly

P Puolakkainen; V Valtonen; A Paananen; T Schröder



Maternal serum ADMA is not associated with proinflammatory cytokines or C-reactive protein during normal pregnancy  

Microsoft Academic Search

Normal pregnancy is associated with changes in the immune system. We studied whether asymmetrical dimethylarginine (ADMA) is associated with this immune system change by assaying high sensitive C-reactive protein (hsCRP), interleukin-6 (IL-6) and tumor necrosis factor-? (TNF-?). The cytokine and dimethylarginine serum concentrations were determined from women with normal pregnancy (n=77) and healthy non-pregnant controls (n=61) matched for age and

Pirjo Valtonen; Kari Punnonen; Heli Saarelainen; Nonna Heiskanen; Olli T. Raitakari; Jorma S. A. Viikari; Tiina Lyyra-Laitinen; Tomi Laitinen; Seppo Heinonen



Application of immunoblotting to serum protein phenotyping with reference to ? 2 HS-glycoprotein (AHS) typing of bloodstains  

Microsoft Academic Search

Sera and bloodstain extracts were subjected to isoelectric focusing in polyacrylamide gels. The focused proteins were transferred to nitrocellulose membranes by diffusion or electrophoretically, then allowed to react with specific antiserum and, after washing, with peroxidase-labeled anti-rabbit IgG. The immune complexes formed on the membranes were detected with 4-chloro-1-naphthol and hydrogen peroxide. Serum group-specific component, a2HS-glycoprotein, the sixth and the

Y. Tamaki; M. Fukuda; H. Nishimukai; T. Kishida



Effect of Bcl2 Expression on Hybridoma Cell Growth in Serum-Supplemented, Protein-Free and Diluted Media  

Microsoft Academic Search

Two transfected hybridoma cell lines TB\\/C3-bcl2 (overexpressing the Bcl-2 protein) and TB\\/C3-pEF (control cell line), were\\u000a compared in batch suspension cultures using a medium supplemented either with horse serum or with a protein-free, iron-rich\\u000a supplement. The membrane intact index (percentage of cells with intact membranes determined by trypan blue staining) of the\\u000a TB\\/C3-bcl2 cell line decreased much slower than that

D. Fassnacht; S. Rössing; F. Fran?k; M. Al-Rubeai; R. Pörtner



Increased serum myeloid-related protein 8\\/14 level is associated with atherosclerosis in type 2 diabetic patients  

Microsoft Academic Search

Background  Myeloid-related protein 8\\/14 (MRP8\\/14) is a stable heterodimer formed by two different calcium-binding proteins (MRP8 and\\u000a MRP14). Studies have identified that MRP8\\/14 regulates vascular inflammation and serves as a novel marker of acute coronary\\u000a syndrome. In this study, we evaluated the correlation between serum levels of MRP8\\/14, hsCRP, endogenous secretory receptor\\u000a for advanced glycation end-products (esRAGE) and the occurrence of

Wen Hui Peng; Wei Xia Jian; Ling Hai Li; Lei Hou; Yi Dong Wei; Wei Ming Li; Ya Wei Xu



Surface-enhanced laser desorption\\/ionization time-of-flight mass spectrometry: serum protein profiling in seminoma patients  

Microsoft Academic Search

Purpose  Surface-enhanced laser desorption ionization time-of-flight mass spectrometry (SELDI-TOF MS) allows rapid protein profiling\\u000a of complex biological mixtures. We analyzed testicular germ cell cancer serum samples to differentiate between cancer and\\u000a controls with a special focus on ?-hCG-negative seminomas.\\u000a \\u000a \\u000a \\u000a \\u000a Methods  Proteomic spectra were generated by the ProteinChip® system and analyzed by the proteomic platform “”. For statistical analysis, an artificial intelligence learning

Romy Strenziok; Stefan Hinz; Christian Wolf; Tim Conrad; Hans Krause; Kurt Miller; Mark Schrader



Calcium dobesilate reduces endothelin-1 and high-sensitivity C-reactive protein serum levels in patients with diabetic retinopathy  

PubMed Central

Purpose To determine the benefits of calcium dobesilate (CaD) administration on endothelial function and inflammatory status in patients with diabetic retinopathy through measurement of serum levels of endothelin-1 and high-sensitivity C-reactive protein (hsCRP). Methods In a double-blind, randomized clinical trial, 90 patients with either severe nonproliferative or proliferative diabetic retinopathy and with blood glucose level of 120–200 mg/dl were randomly allocated to treatment with either CaD tablets (500 mg daily) or placebo for 3 months. Visual acuity, intraocular pressure, and macular status were performed before the study. The serum levels of endothelin-1 and hsCRP were evaluated in both groups before and at the third month of the trial. Results The median serum level of hsCRP significantly differed between the groups 3 months following the CaD or placebo administration (2.2 mg/l in the CaD group versus 3.7 mg/l in the placebo group, p=0.01). The mean endothelin-1 serum level was 0.69±0.32 pg/ml in the CaD group and 0.86±0.30 pg/ml in the placebo group (p=0.01). Furthermore, in the CaD group, the serum levels of both endothelin-1 and hsCRP were significantly decreased 3 months after administration of CaD (p<0.001). Conclusions Administration of the CaD in the patients with diabetic retinopathy may reduce the serum levels of endothelin-1 and hsCRP. This might imply amelioration of the endothelial function and inflammatory status following CaD therapy in these patients.

Javadzadeh, Alireza; Ghorbanihaghjo, Amir; Adl, Farzad Hami; Andalib, Dima; Khojasteh-Jafari, Hassan



Granulocyte colony-stimulating factor induces Egr-1 up-regulation through interaction of serum response element-binding proteins.  


Granulocyte colony-stimulating factor (G-CSF) stimulates the proliferation and maturation of myeloid progenitor cells both in vitro and in vivo. We showed that G-CSF rapidly and transiently induces expression of egr-1 in the NFS60 myeloid cell line. Transient transfections of NFS60 cells with recombinant constructs containing various deletions of the human egr-1 promoter identified the serum response element (SRE) between nucleotides (nt) -418 and -391 as a critical G-CSF-responsive sequence. The SRE (SRE-1) contains a CArG box, the binding site for the serum response factor (SRF), which is flanked at either side by an ETS protein binding site. We demonstrated that a single copy of the wild-type SRE-1 in the minimal promoter plasmid, pTE2, is sufficient to induce transcriptional activation in response to G-CSF and that both the ETS protein binding site and the CArG box are required for maximal transcriptional activation of the pTE2-SRE-1 construct. In electromobility shift assays using NFS60 nuclear extracts, we identified SRF and the ETS protein Fli-1 as proteins that bind the SRE-1. We also demonstrated through electrophoretic mobility shift assays, using an SRE-1 probe containing a CArG mutation, that Fli-1 binds the SRE-1 independently of SRF. Our data suggest that SRE-binding proteins potentially play a role in G-CSF-induced egr-1 expression in myeloid cells. PMID:10806199

Mora-Garcia, P; Sakamoto, K M



Reactivity of anticancer metallodrugs with serum proteins: new insights from size exclusion chromatography-ICP-MS and ESI-MS.  


A method based on the coupling of high resolution size-exclusion liquid chromatography using a polymer stationary phase with inductively coupled plasma mass spectrometry was developed to study the interactions of two metallodrugs - cisplatin and RAPTA-T - with the serum proteins albumin and transferrin. In contrast to previous approaches, the technique allowed the total recovery of the metals from the column and was able to discriminate between the different species of the metallodrugs and their complexes with the proteins at femtomolar detection levels. Metal binding was found to be dependent on the protein concentration and on the incubation time of the sample. Cisplatin was found to bind the serum proteins to the same extent, whereas RAPTA-T showed marked preference for transferrin. The affinity of the ruthenium complex for holo-transferrin was higher than for the apo-form suggesting a cooperative iron-mediated metal binding mechanism. RAPTA-T binding to holo-transferrin was further investigated by electrospray mass spectrometry using both the intact protein and a model peptide mimicking the iron-binding pocket. PMID:21151827

Groessl, Michael; Terenghi, Mattia; Casini, Angela; Elviri, Lisa; Lobinski, Ryszard; Dyson, Paul J



Identification of Tumor-associated Autoantigens for the Diagnosis of Colorectal Cancer in Serum Using High Density Protein Microarrays*  

PubMed Central

There is a mounting evidence of the existence of autoantibodies associated to cancer progression. Antibodies are the target of choice for serum screening because of their stability and suitability for sensitive immunoassays. By using commercial protein microarrays containing 8000 human proteins, we examined 20 sera from colorectal cancer (CRC) patients and healthy subjects to identify autoantibody patterns and associated antigens. Forty-three proteins were differentially recognized by tumoral and reference sera (p value <0.04) in the protein microarrays. Five immunoreactive antigens, PIM1, MAPKAPK3, STK4, SRC, and FGFR4, showed the highest prevalence in cancer samples, whereas ACVR2B was more abundant in normal sera. Three of them, PIM1, MAPKAPK3, and ACVR2B, were used for further validation. A significant increase in the expression level of these antigens on CRC cell lines and colonic mucosa was confirmed by immunoblotting and immunohistochemistry on tissue microarrays. A diagnostic ELISA based on the combination of MAPKAPK3 and ACVR2B proteins yielded specificity and sensitivity values of 73.9 and 83.3% (area under the curve, 0.85), respectively, for CRC discrimination after using an independent sample set containing 94 sera representative of different stages of progression and control subjects. In summary, these studies confirmed the presence of specific autoantibodies for CRC and revealed new individual markers of disease (PIM1, MAPKAPK3, and ACVR2B) with the potential to diagnose CRC with higher specificity and sensitivity than previously reported serum biomarkers.

Babel, Ingrid; Barderas, Rodrigo; Diaz-Uriarte, Ramon; Martinez-Torrecuadrada, Jorge Luis; Sanchez-Carbayo, Marta; Casal, J. Ignacio



The role of recombinant proteins in the developmentof serum-free media  

Microsoft Academic Search

Early developments in serum-free media led to a variety of formulations in which components normally provided in serum and\\u000a required for growth (insulin, transferrin, lipid supplements, trace elements) and poorly defined components (extracts, hydrolysates)\\u000a were added to defined basal media. These additives were mostly animal-derived. Given recent concerns about TSEs (transmissible\\u000a spongiform encephalopathies) and other adventitious agents, the drive in

Joanne Keenan; Dermot Pearson; Martin Clynes



Meta-analysis of the effect of initial serum protein concentration and empirical prediction model for growth of neonatal Holstein calves through 8 weeks of age.  


A data set was constructed from individual calf means gathered in the Nurture Research Center (Lewisburg, OH) and used in a meta-analysis to parameterize an empirical model predicting growth measures for neonatal calves. The data set contained 993 observations from 20 research trials conducted in all seasons of multiple years. Growth measures gathered included average daily gain (ADG) preweaning, postweaning, and through 8 wk of age. Independent variables gathered included age at weaning; total starter intake (SI); total milk replacer intake (MRI); milk replacer CP (MRCP) and fat (MRfat) contents; number of days with abnormal fecal scores (AFS); average environmental temperature preweaning, postweaning, and through 8 wk of age; minimum and maximum temperature during the entire 8 wk; body weight at d 0; and initial serum protein concentration. Additionally, the interactions of SI, MRI, and MRCP and MRfat were considered for the model. Backward elimination multiple regressions were conducted using a mixed model with a random effect of trial. The final model for total ADG indicated that increasing SI or MRI improves calf growth. Also, increasing MRCP or MRfat increased growth. Increased sickness (as measured by increased AFS) or increased body weight at d 0 decreased ADG. Growth of neonatal dairy calves appears to be more controlled by nutrient intake and their interactions than by surrogates for health status of the calves (AFS and initial serum protein concentration) or environmental temperature. PMID:22192215