Identification of Unique Biomarkers in Sepsis Associated Coagulopathy

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 1237-1237
Author(s):  
Gautam Sharma ◽  
Rahul Bijlani ◽  
Evangelos Litinas ◽  
Debra Hoppensteadt ◽  
Josephine Cunanan ◽  
...  
Keyword(s):  
Conflicts Of Interest ◽  
Protein Chip ◽  
Plasma Samples ◽  
Molecular Weight Range ◽  
Array Technology ◽  
Surface Enhanced ◽  
Apparent Reduction ◽  
Normal Humans ◽  
Normal Human ◽  
Pathologic Processes

Abstract Abstract 1237 Introduction: Disseminated Intravascular Coagulation represents a complex multi-factorial pathophysiological process in which protease disregulation along with other pathologic processes play an important role. The uncontrolled protease activities result in generation of several mediators which are not fully characterized. Biomarker profiling using protein chip array technology has been used in the identification of unique mediators in various diseases. Surface Enhanced Laser Desorption Ionisation(SELDI) is an ionization method in mass spectrometry that is used for the analysis of protein mixtures and correspondingly in the profiling of biomarkers in various diseases. Purpose: The purpose of this study was to profile and identify unique biomarkers in hospitalized patients with sepsis associated coagulopathy. Study design: There were 385 plasma samples included in this study. Baseline protein chip array profile in Molecular weight range of 0–200 kDa was carried out using SELDI technique employing gold chips. Plasma samples from normal humans (n=100) and pooled preparation from both the normal (NHP) and suspect liver disease/coagulopathy were utilized as controls. Results: Of the 385 baseline samples analyzed 371 patients (96.86%) exhibited the presence of a unique biomarker at 11.6 kDa. In addition down regulation of a biomarker at 56.4 kDa was also noted in 218 of patients (56.74%) In the normal human pooled plasma the 11.6 kDa peak was nearly absent. However in Pathological pooled plasma a distinct biomarker at 11.6 kDa with a much lower intensity was noted. Additional markers in baseline samples in 1–10 kDa range were also noted. Conclusions: These observations suggest that sepsis associated coagulopathy results in the generation of specific unique biomarker at 11.6kDa which requires further characterization. The downregulation of biomarker at 56.4 kDa and the apparent reduction levels during hospitalization require further investigations. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Ecological Effect of Ceftaroline-Avibactam on the Normal Human Intestinal Microbiota

2015 ◽  
Vol 59 (8) ◽  
pp. 4504-4509 ◽  
Author(s):  
Mamun-Ur Rashid ◽  
Staffan Rosenborg ◽  
Georgios Panagiotidis ◽  
Karin Söderberg-Löfdal ◽  
Andrej Weintraub ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Intestinal Microbiota ◽  
Ecological Effect ◽  
New Combination ◽  
Plasma Samples ◽  
Ceftaroline Fosamil ◽  
Content Type ◽  
Human Intestinal Microbiota ◽  
Normal Human ◽  
Lactamase Inhibitor

ABSTRACTCeftaroline-avibactam is a new combination of the antibiotic ceftaroline with a novel non-β-lactam β-lactamase inhibitor, avibactam. The purpose of the present study was to investigate the effect of ceftaroline-avibactam on the human intestinal microbiota. Fourteen healthy volunteers received ceftaroline-avibactam (600 mg ceftaroline fosamil and 600 mg avibactam) intravenously over 2 h every 8 h on days 1 to 6 and as a single dose on day 7. Fecal samples were collected on day −1 (within 24 h of the first infusion on day 1) and on days 2, 5, 7, 9, 14, and 21.Escherichia colinumbers decreased during the study and normalized on day 21. An increased number ofKlebsiellabacteria appeared on day 14 and normalized on day 21. The number of other enterobacteria decreased during the study, and the number of enterococci decreased from days 2 to 7 and normalized on day 9.Candidanumbers increased from days 5 to 9 and normalized after day 14. The number of lactobacilli decreased during the study and recovered on day 14. The number of bifidobacteria decreased on day 2 and normalized on day 21. The number ofBacteroidesbacteria was unchanged.Clostridium difficilenumbers decreased on days 7 and 9 and increased on days 14 and 21. A toxigenicC. difficilestrain was detected in one volunteer on day 21 with no reported adverse events. Plasma samples were collected on days −1, 2, 5, and 7. Ceftaroline and avibactam concentrations were 0 to 34.5 mg/liter and 0 to 61.6 mg/liter, respectively, in plasma and 0 to 35.4 mg/kg and 0 to 98.5 mg/kg, respectively, in feces. (This study is registered in the European Clinical Trials Database [https://eudract.ema.europa.eu/] under number EudraCT 2012 004921-25.)

The Determination of Triglycerides in Plasma and Tissues

1968 ◽  
Vol 14 (2) ◽  
pp. 156-161 ◽  
Author(s):  
Vishwanath M Sardesai ◽  
Joan A Manning
Keyword(s):  
Human Subjects ◽  
Plasma Samples ◽  
Simple Method ◽  
Glycerol Moiety ◽  
Normal Human ◽  
Time Required ◽  
Colored Compound

Abstract A simple method for the determination of plasma and tissue triglycerides is described. This procedure involves the extraction and saponification of triglycerides, the oxidation of the glycerol moiety to formaldehyde, and the conversion of formaldehyde to a yellow-colored compound, 3,5 diacetyl-1-4 dihydrolulidine, the intensity of which is determined spectrophotometrically. The recoveries of triglycerides added to plasma and tissues have been satisfactory. Plasma samples obtained from normal human subjects are found to have triglycerides in the range 83-200 mg./100 ml. From the standpoint of sensitivity, simplicity, and time required, this technic is believed to be an improvement over previously described procedures for triglyceride determination.

scholarly journals Effects of fresh-frozen plasma and its cryosupernatant fraction on von Willebrand factor multimeric forms in chronic relapsing thrombotic thrombocytopenic purpura

Blood ◽  
1985 ◽  
Vol 65 (5) ◽  
pp. 1232-1236 ◽  
Author(s):  
JL Moake ◽  
JJ Byrnes ◽  
JH Troll ◽  
CK Rudy ◽  
SL Hong ◽  
...  
Keyword(s):  
Plasma Exchange ◽  
Fresh Frozen Plasma ◽  
Plasma Samples ◽  
Fresh Frozen ◽  
Normal Human ◽  
Von Willebrand ◽  
Willebrand Factor ◽  
Frozen Plasma

Abstract Remission plasma samples of some patients with chronic relapsing thrombotic thrombocytopenic purpura (TTP) contain unusually large von Willebrand factor (vWF) multimers similar to those produced by normal human endothelial cells in culture. The infusion of the cryosupernatant fraction of normal plasma is as effective as normal fresh-frozen plasma (FFP) in the treatment or prevention of TTP episodes in patients with the chronic relapsing form of TTP. Three patients with chronic relapsing TTP during remission have unusually large vWF multimers present in their plasma. Two of the patients were transfused once with FFP, one of the two received cryosupernatant on three occasions, and the third patient was studied before and immediately after plasma exchange. Unusually large vWF multimers decreased or disappeared from patient plasma samples within 1/2 to 1 1/2 hours following the transfusion of FFP (on two occasions) or cryosupernatant (on two of three occasions), and immediately after plasma exchange (on one occasion). The patient who received cryosupernatant was studied serially after the infusions. Unusually large vWF multimers returned to her plasma within ten to 24 hours and persisted thereafter. Unusually large vWF multimers did not disappear from patient remission plasma samples, or from the culture medium removed from normal human endothelial cells, when these fluids were incubated in vitro with either normal FFP or cryosupernatant. We conclude that an activity in FFP, and its cryosupernatant fraction, promoted the rapid in vivo disappearance of unusually large vWF multimers from the plasma of two patients with chronic relapsing TTP in remission, and plasma exchange reversed the abnormality in a third patient who was in partial remission. Neither FFP nor cryosupernatant directly converted unusually large multimers to smaller vWF forms in vitro in the fluid phase. These results indicate that an activity in the cryosupernatant fraction of normal plasma is involved in vivo in controlling the metabolism of unusually large vWF multimers, and that this process is defective in some chronic relapsing TTP patients.

Molecular Heterogeneity in Recombinant Erythropoietin Using Surface Enhanced Laser Induced Desorption Ionization (SELDI). Clinical Implications.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 3557-3557
Author(s):  
E. James ◽  
J. Maddineni ◽  
J. Fareed ◽  
M. Florian-Kujawski ◽  
F. Baltasar ◽  
...  
Keyword(s):  
Recombinant Proteins ◽  
Protein Chip ◽  
Protein Profiling ◽  
Molecular Profile ◽  
Molecular Heterogeneity ◽  
Recombinant Erythropoietin ◽  
Heterogeneous Distribution ◽  
Array Technology ◽  
Protein Components ◽  
Main Component

Abstract Several erythropoietin preparations (EPO) are used globally for anemia management. Repeated EPO use has been reportedly associated with thrombotic complications. Molecular profiles of these recombinant proteins vary widely, impacting their clinical profile. This may be due to cellular/receptor interactions and other modulatory properties. There are WHO and FDA regulatory guidelines regarding molecular profile; data on protein profiling is usually not provided by the manufacturers. Purpose of this investigation was to compare various clinically-used recombinant erythropoietin products Epogen (Amgen), Vintor (Emcure), Eprex (Janssen-CILAG) and Neo Recormon (Roche) utilizing protein chip array technology (Ciphergen, Fremont, CA). All products were analyzed using anionic SAX2 chips and molecular profile was obtained in the range of 0–150 KDa. Epogen and Vintor exhibited a main component at 32.4 KDa range with minor peaks at 13.5, 21.7 and 43.2 KDa. Intensity of the main component varied widely even within the 3 batches of Epogen. The Epogen preparations and Vintor brand contained albumin in different quantities. One batch of Epogen contained 4x higher amount of this material with a heterogeneous distribution. Epex and Neo Recormon did not have any albumin. One of the Epogen product also contained a component at 110.0 KDa. Glycosylation profiles of each of these preparations also differ. These data suggest that molecular heterogeneity exists within different recombinant erythropoietin preparations. Molecular heterogeneity among these preparations may be partly responsible for differential interactions within erythropoietin receptors on cells and anti-epo antibody generation. Protein chip array technology may be helpful in examining the protein components within different recombinant erythropoietins and may be helpful in establishing molecular profile guideline to minimize pharmacodynamic variance in erythropoietin preparations. Furthermore, molecular profiling of these recombinant proteins may be helpful in establishing the purity and homogeneity of these drugs.

Upregulation of Inflammatory Mediators In End-Stage Renal Disease as Measured Via Biochip Array Technology

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 5186-5186
Author(s):  
Vinod Bansal ◽  
Rachael Davis ◽  
Evangelos Litinas ◽  
Debra Hoppensteadt ◽  
Indermohan Thethi ◽  
...  
Keyword(s):  
Renal Disease ◽  
Inflammatory Mediators ◽  
End Stage Renal Disease ◽  
Conflicts Of Interest ◽  
Fold Increase ◽  
Endothelial Damage ◽  
Cellular Damage ◽  
Array Technology ◽  
Stage Renal Disease ◽  
End Stage

Abstract Abstract 5186 End-Stage Renal Disease (ESRD) is a complex syndrome in which systemic vascular pathophysiologic changes contribute to adverse cardiovascular and cerebrovascular manifestations. Cardiovascular disease alone is present in over 60% of patients with ESRD and contributes heavily to mortality among this population. Given that inflammatory and hemostatic aberrations contribute to the overall pathogenesis of the syndrome, the purpose of this study is to profile several inflammatory mediators in order to better understand their role in the underlying mechanism of vascular changes in ESRD. Plasma samples from 49 patients with ESRD were collected prior to maintenance hemodialysis sessions. A group of 56 normal individuals, both male and female, was included as control. Cerebral Array II chips were used in the Randox® system to simultaneously measure Neuron Specific Enolase (NSE), Neutrophil Gelatinase-associated Lipocalin (NGAL), Soluble Tumor Necrosis Factor Receptor I (TNFRI), D-Dimer (DD), Thrombomodulin (TM), and C-reactive protein (CRP). The Randox® Evidence Investigator™ is a new biochip array technology that utilizes multiple discrete test regions of immobilized antibody to simultaneously quantify multiple markers from a single patient plasma sample based on the light signal generated from each test region. The data was statistically analyzed using the Mann-Whitney U test (two-tailed with Gaussian approximation). As compared to the normal individual, all of the markers studied showed an upregulation in patients with ESRD. Most notably, TNFRI showed a 19.8 fold increase in patients with ESRD (mean 7.8 ± 2.8 ng/ml, range 0.8 to 13.7) compared to the control (mean 0.4 ± 0.2, range 0.1 to 1.0). TM was increased 5.2 fold (mean 6.5 ± 2.6, range 0.7 to 14.1) compared to control (mean 1.2 ± 0.4, range 0.6 to 2.3). Also, NGAL showed a 4.6 fold increase (mean 1390 ± 257, range 406 to 1729), compared to control (mean 299 ± 99, range 115 to 603), and CRP a 4.2 fold increase (mean 5.7 ± 4.2 ug/ml, range 0.6 to 13.2) compared to control (mean 1.4 ± 1.7, range 0.2 to 11.4). DD and NSE were also increased 3.0 and 1.8 fold respectively. These studies show that some newer markers such as TNFRI, NGAL and NSE are upregulated in ESRD. The marked increase in TM is highly suggestive of endothelial damage. Similarly, the increase in TNFRI supports a state of increased cellular damage. The elevations in NGAL and CRP imply a state of increased inflammation and indicate a polypathologic process, which may predispose ESRD patients to both cardiovascular and cerebrovascular thromboembolic events. Finally, this study further validates the role of endothelial damage and endogenous thrombotic processes in ESRD as evidenced by the increased levels of TM and DD. However, the clinical significance of these markers still needs to be further explored. Disclosures: No relevant conflicts of interest to declare.

Comparison of Branded Enoxaparin (Lovenox) and a Biosimilar Version of Enoxaparin (Fibrinox)

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 4385-4385
Author(s):  
Walter Jeske ◽  
Elizabeth McGeehan ◽  
Omer Iqbal ◽  
Debra Hoppensteadt ◽  
Jeanine M. Walenga ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Thrombin Generation ◽  
Conflicts Of Interest ◽  
Average Molecular Weight ◽  
Molecular Profile ◽  
Distribution Analysis ◽  
Thrombin Time ◽  
Plasma Samples ◽  
Branded Product ◽  
Inhibition Of Thrombin

Abstract Abstract 4385 Several biosimilar versions of branded enoxaparin (Lovenox, Sanofi-Aventis, Paris, France) have recently become available throughout the world. These biosimilar enoxaparin preparations are distributed by multiple suppliers in Asia and in North and South America. Enoxaparin represents a complex mixture of oligosaccharides obtained by alkaline depolymerization of porcine mucosal heparin. It is the most widely used low molecular weight heparin which has been validated for clinical use in multiple indications. While the molecular profile and anti-Xa potencies of some of the biosimilar versions of enoxaparin are comparable, product based differences have been reported amongst some of the biosimilar versions of enoxaparin. The purpose of this study was to compare the biochemical and pharmacologic profile of one biosimilar version of enoxaparin, namely Fibrinox (Sandoz SA, Buenos Aires, Argentina) with the branded product Lovenox. The products were compared in equigravimetric amounts, assuming equivalent potency (100 AXa U/mg). Both products exhibited comparable molecular weight profiles in terms of average molecular weight and oligosachharide distribution. Analysis of the antithrombin binding hexasaccharide fractions of Fibrinox and Lovenox indicated the presence of eight distinct hexasaccharides. The relative proportions these hexasaccharides differed between Fibrinox and Lovenox. The anti-Xa and anti-IIa activities were comparable. In the whole blood clot-based assays such as TEG and ACT, both agents produced similar anticoagulant effects. In the plasma based assays such as the APTT, Heptest and thrombin time, both products showed comparable anticoagulant effects in the normal human pooled plasma samples. However, in plasma samples collected from patients with liver disease who were apparently anticoagulant free, the two products showed differences in their anticoagulant effects in the APTT assay (p<0.05). In the TF mediated thrombin generation assay, Fibrinox produced a stronger inhibition of thrombin generation compared to Lovenox (IC50; Fibrinox, 1.6 μ g/ml, Lovenox 2.2 μ g/ml). No differences were observed between the two products in the agonist induced platelet aggregation assays. However in the 14C serotonin release study, Fibrinox produced a stronger HIT serum mediated 14C release (p<0.05). Differences in the fibrinokinetic profile and the inhibition of thrombin activatable fibrinolytic inhibitor activation were observed with these LMWHs. These studies suggest while both the molecular profile and the pharmacopoeial potency of Fibrinox is similar to the branded product, these drugs can be differentiated in some of the other assays and should be evaluated in terms of additional pharmacologic mechanisims to demonstrate bioequivalence. Disclosures: No relevant conflicts of interest to declare.

MicroRNA Maestros of Hematopoiesis

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. SCI-32-SCI-32
Author(s):  
Kara A. Scheibner ◽  
Diane Heiser ◽  
Ian M Kaplan ◽  
Wen-Chih Cheng ◽  
MinJung Kim ◽  
...  
Keyword(s):  
Target Genes ◽  
Conflicts Of Interest ◽  
Loss Of Function ◽  
Hematopoietic Stem ◽  
Cell Functions ◽  
Human Cd34 ◽  
Leukemia Treatment ◽  
Normal Human ◽  
Target Molecules ◽  
And Function

Abstract Abstract SCI-32 MicroRNAs (miRs) inhibit stability and/or translation of mRNAs, usually by binding to specific sites in the 32′UTRs of their target mRNAs. Due to imperfect (i.e. partially complementary) miR:mRNA base-pairing, miRs can block translation of many mRNAs and serve as powerful master switches to regulate cell functions. Therefore, we profiled miR expression in human CD34+ hematopoietic stem-progenitor cells (HSPCs) and combined human HSPC miR expression, mRNA expression, and miR-mRNA target predictions to hypothesize that certain HSPC-expressed miRs (HE-miRs) target several mRNAs critical to hematopoiesis. On this informatic basis, we formulated a model of hematopoietic differentation in which many genes specifying hematopoietic differentiation are expressed by early HSPCs, but held in check by miRs [1]. In addition, we noted that the miR-23a cluster (i.e. adjacent, co-transcribed miR-23a, miR-27a, and miR-24-2) is not expressed or is expressed at levels >2-fold lower in 50% of acute myeloid leukemias and 80% of acute lymphoid leukemias tested compared to normal human HSPCs. ‘Re-expressing’ 1 or more of these miR-23a cluster members in leukemia cells promotes their apoptosis and reduces their proliferation, thus suggesting that these miRs have a tumor suppressive role. We have identified YWHAQ (14-3-3q) and several other 14-3-3 isoforms, which are anti-apoptotic and have established roles as oncogenes, as miR-23a cluster target molecules. Artificial manipulation of these HE-miRs and their target genes may lead to novel strategies for leukemia treatment and/or for expansion of normal HSPCs. Since the CD34+ HSPCs that we studied initially include rare stem cells and various stages of progenitors, we have expanded our miR profiling to more highly purified subsets of mouse HSPCs. Several previously described (e.g. miR-155 [1], miR-451 [2], miR-146 [3]) and novel HE-miRs are expressed differentially in lineages/stages of HSPCs, and their selective expression has been confirmed in human HSPC subsets. We are using cellular gain- and loss-of-function approaches with hematopoietic functional assays to determine whether these HE-miRs control human hematopoiesis. Understanding the effects of HE-miRs in hematopoiesis may elucidate hematopoietic and general stem cell biologic mechanisms. 1. Georgantas RW, 3rd, Hildreth R, Morisot S, Alder J, Liu CG, Heimfeld S, Calin GA, Croce CM, Civin CI. CD34+ hematopoietic stem-progenitor cell microRNA expression and function. A circuit diagram of differentiation control. Proc Natl Acad Sci USA. 2007;104:2750–2755. 2. Dore LC, Amigo JD, Dos Santos CO, Zhang Z, Gai X, Tobias JW, Yu D, Klein AM, Dorman C, Wu W, Hardison RC, Paw BH, Weiss MJ. A GATA-1-regulated microRNA locus essential for erythropoiesis. Proc Natl Acad Sci USA. 2008;105:3333–3338. 3. Starczynowski DT, Kuchenbauer F, Argiropoulos B, Sung S, Morin R, Muranyi A, Hirst M, Hogge D, Marra M, Wells RA, Buckstein R, Lam W, Humphries RK, Karsan A. Identification of miR-145 and miR-146a as mediators of the 5q- syndrome phenotype. Nat Med. 2010;16:49–58. Disclosures: No relevant conflicts of interest to declare.

Monitoring ADAMTS13 Autoantibody Titer Using a Novel Quantitative Radioimmunoprecipitation Assay In Patients With Acquired Thrombotic Thrombocytopenic Purpura

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 3561-3561
Author(s):  
Atsuko Igari ◽  
Takanori Moriki ◽  
Hideo Wada ◽  
Kenji Soejima ◽  
Mitsuru Murata
Keyword(s):  
Monoclonal Antibodies ◽  
Plasma Exchange ◽  
Thrombotic Thrombocytopenic Purpura ◽  
Conflicts Of Interest ◽  
Expression System ◽  
Plasma Samples ◽  
Thrombocytopenic Purpura ◽  
Free Protein ◽  
Autoantibody Titer ◽  
Radioimmunoprecipitation Assay

Abstract Background ADAMTS13-binding immunoglobulin G (IgG)-type autoantibodies are present in patients with acquired thrombotic thrombocytopenic purpura (TTP), thereby causing severe deficiency of plasma ADAMTS13 activity. Some specific autoantibodies directly inhibit the enzymatic activity by interfering with the access of its substrate, von Willebrand factor (VWF), particularly in the spacer domain, although others bind to almost all of the domains in the molecule, possibly leading to the acceleration of the clearance from blood stream. Not only the inhibitor titer, but the importance of ADAMTS13 autoantibody titer was highlighted by many previous clinical studies, associating with the prognosis such as recurrence rate. Objective We targeted to establish a novel high-sensitive assay to measure ADAMTS13-binding IgG autoantibody titer using three kinds of radioisotope-labeled antigens, ADAMTS13 whole molecule, MDTCS and T2-8/CUB, and aimed to analyze the association between the autoantibody titer and the clinical characteristics of TTP patients. Materials and Methods Human Cell-Free Protein Expression System (Takara #3281, Shiga, Japan) was used to synthesize radioisotope-labeled antigens. ADAMTS13 cDNA corresponding to whole molecule (A13), metalloprotease to spacer domains (MDTCS) and TSP1-2 to CUB2 domains (T2-8/CUB) were cloned into an expression vector pT7-IRES and mixed with Cell Lysate containing T7 RNA polymerase, methionine-free amino acids, ATP, translation enhancement factor and 35S-methionine. The correct synthesis and molecular size of the radiolabeled antigens were checked with SDS-PAGE. To assess the utility as a quantitative assay, each of the antigens was mixed with mouse anti-ADAMTS13 monoclonal antibodies, whose epitopes were determined in our previous study (Thromb Res. 2012; 130(3):e79-83), and the immune complex was precipitated with protein G beads, washed and measured in a liquid scintillation counter. Plasma samples from acquired TTP patients were tested to quantify the autoantibody titers using radiolabeled A13, MDTCS and T2-8/CUB antigens, respectively. As a control, plasma samples from healthy subjects with no histories of autoimmune disease were also tested. Results Each of the radiolabeled antigens was detected as a single band at the correct molecular weight size and successfully immnoprecipitated with several mouse anti-ADAMTS13 monoclonal antibodies, indicating the intact molecular conformation of the synthesized proteins using the cell-free protein synthesis system. Moreover, the appropriate dose-dependent escalation curves in accordance with the addition of the monoclonal antibodies were observed, thereby confirming the utility of the assay as a quantitative analysis. We tested TTP patient plasma at onset (n=5) and were able to detect ADAMTS13 autoantibody titers with each of the radiolabeled A13, MDTCS and T2-8/CUB antigens. We next applied this assay for monitoring ADAMTS13 autoantibody titer in a clinical course of TTP patient. The patient developed the first episode of TTP at the age of 2 month and treated with steroid pulse and plasma exchange therapy for six consecutive days. Remission was once achieved but 6 months later from the onset, the second episode of TTP occurred and the patient was treated with 11 plasma exchange and rituximab at the dose of 375 mg/m2 once a week for 4 weeks. Plasma samples at onset, after the first 6 consecutive plasma exchange and after rituximab administration were examined about autoantibody titer using this assay. Interestingly, the titers remained high even after the plasma exchange but declined clearly after the rituximab treatment, whereas no reduction of total IgG level was observed. These findings suggest that the autoantibody titration using this assay might be useful to assess the effect of treatment and associate with the prognosis related to the recurrence. Conclusion We developed a novel quantitative radioimmunoprecipitation assay to measure ADAMTS13 autoantibody titer related to three antigens, ADAMTS13 whole molecule, MDTCS and TSP2-8/CUB. This assay may serve not only as a diagnostic test but as a monitoring index to evaluate the prognosis of TTP. Disclosures: No relevant conflicts of interest to declare.

AN INSULIN ASSAY BASED ON THE INCORPORATION OF LABELLED GLYCINE INTO PROTEIN OF ISOLATED RAT DIAPHRAGM

1959 ◽  
Vol 19 (3) ◽  
pp. 259-262 ◽  
Author(s):  
K. L. MANCHESTER ◽  
P. J. RANDLE ◽  
F. G. YOUNG
Keyword(s):  
Human Plasma ◽  
Plasma Samples ◽  
Rat Diaphragm ◽  
Normal Human Plasma ◽  
Advantages And Disadvantages ◽  
Straight Line ◽  
Normal Human ◽  
Isolated Rat

SUMMARY Insulin increases the incorporation of [14C]glycine into the protein of isolated rat diaphragm in vitro. This effect of the hormone can be used as the basis of a straight-line assay for insulin. The advantages and disadvantages of such a method of assay are discussed. Normal human plasma also stimulates incorporation of glycine into diaphragm protein, and the activities of two plasma samples were equivalent to 2 and 10 mu. insulin/ml. plasma, respectively.

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