Discrimination between intact and mid-C-regional PTH using selective radioimmunoassay systems

1983 ◽  
Vol 102 (4) ◽  
pp. 543-548 ◽  
Author(s):  
H. Jüppner ◽  
M. Rosenblatt ◽  
G. V. Segre ◽  
R.-D. Hesch
Keyword(s):  
Renal Insufficiency ◽  
Secondary Hyperparathyroidism ◽  
Gel Filtration ◽  
Assay System ◽  
Intact Pth ◽  
Peritoneal Dialysate ◽  
High Affinity ◽  
Maximal Displacement ◽  
Pth Radioimmunoassay

Abstract. Two 125I-labelled hormone preparations (synthetic 44–68(Tyr)hPTH bearing a tyrosin at position No. 44 and 1–84 bPTH) were used to characterize antiserum G III for PTH radioimmunoassay. PTH fragments were used for displacement of either tracer and included 1–34 hPTH, 28–48 bPTH, 25–48 bPTH, 28–54 hPTH, 32–43 hPTH, 42–55(Tyr)hPTH (tyrosin at position No. 42), 1–65 bPTH fragments and intact 1–84 hPTH. Standard curves of the 44–68 hPTH, 1–65 bPTH and the 1–84 hPTH run in parallel to each other independent of the tracer used for the assay. When using the 44–68(Tyr)hPTH tracer, half-maximal displacement (1/2 max) was achieved at 24 pg/tube for 44–68 hPTH, 450 pg/tube for 1–65 bPTH and at 1300 pg/tube for 1–84 hPTH. When carrying out the assay with the 1–84 bPTH tracer, the values for 1/2 max were 11.5 pg/tube for 44–68 hPTH, 135 pg/tube for 1–65 bPTH and 370 pg/tube for 1–84 hPTH. Gel filtration of plasma and peritoneal dialysate from one patient with terminal renal insufficiency and secondary hyperparathyroidism indicate that due to the high affinity of the antibody for the mid-C-regional hPTH, this assay system is appropriate for the discrimination of intact PTH and large mid-C-regional fragments, that include most of the 44–68 hPTH sequence.

scholarly journals Characterization of the binding of plasminogen activators to plasma membranes from human liver

1992 ◽  
Vol 287 (3) ◽  
pp. 911-915 ◽  
Author(s):  
G Nguyen ◽  
S J Self ◽  
C Camani ◽  
E K O Kruithof
Keyword(s):  
Human Liver ◽  
Plasma Membranes ◽  
Gel Filtration ◽  
Mannose Receptor ◽  
Hepatoma Cells ◽  
Tissue Type ◽  
High Affinity ◽  
Liver Membranes ◽  
Membrane Bound ◽  
Pai 1

The binding of tissue-type plasminogen activator (t-PA) to membranes prepared from human liver was investigated, and a specific, saturable, high-affinity binding site (Kd = 3.4 nM) was identified. The binding of t-PA to liver membranes was not affected by an excess of D-mannose or D-galactose, or by active urokinase (u-PA), whereas binding of t-PA to membranes prepared from human HepG2 hepatoma cells was inhibited by u-PA. HepG2-membrane-bound t-PA was fully complexed to PA inhibitor 1 (PAI-1), whereas liver-membrane-bound t-PA was not complexed. Gel filtration on Sephacryl S300 of membrane proteins solubilized in deoxycholate revealed that high-affinity t-PA binding activity elutes at an apparent molecular mass of 40 kDa. Monoclonal antibodies specific for the growth factor and the kringle 2 domains inhibited the binding of t-PA to liver membranes and the catabolism of t-PA by rat hepatoma cells. Human liver membranes also bound u-PA; binding was inhibited by pro-u-PA, the N-terminal fragment of u-PA, but not by the 33 kDa form of u-PA or by t-PA. Our results show that human liver membranes contain a specific 40 kDa binding protein for t-PA that is different from the PAI-1-dependent receptor described on HepG2 cells and the mannose receptor isolated from human liver.

scholarly journals Intact PTH assay overestimates true 1-84 PTH levels after maxacalcitol therapy in dialysis patients with secondary hyperparathyroidism

2004 ◽  
Vol 19 (4) ◽  
pp. 892-897 ◽  
Author(s):  
J. J. Kazama ◽  
K. Omori ◽  
N. Higuchi ◽  
N. Takahashi ◽  
Y. Ito ◽  
...  
Keyword(s):  
Secondary Hyperparathyroidism ◽  
Dialysis Patients ◽  
Intact Pth

scholarly journals Pre-operative Localisation of the Parathyroid Glands in Secondary Hyperparathyroidism: A Retrospective Cohort Study

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Takahisa Hiramitsu ◽  
Toshihide Tomosugi ◽  
Manabu Okada ◽  
Kenta Futamura ◽  
Makoto Tsujita ◽  
...  
Keyword(s):  
Computed Tomography ◽  
Cohort Study ◽  
Parathyroid Hormone ◽  
Secondary Hyperparathyroidism ◽  
Retrospective Cohort ◽  
Parathyroid Glands ◽  
Imaging Modalities ◽  
Intact Parathyroid Hormone ◽  
Intact Pth ◽  
Persistent Hyperparathyroidism

Abstract Complete parathyroidectomy (PTx) is essential during total PTx for secondary hyperparathyroidism (SHPT) to prevent recurrent and persistent hyperparathyroidism. Pre-operative imaging evaluations, including computed tomography (CT), ultrasonography (US), and Tc-99m sestamibi (MIBI) scans, are commonly performed. Between June 2009 and January 2016, 291 patients underwent PTx for SHPT after pre-operative evaluations involving CT, US, and MIBI scans, and the diagnostic accuracies of these imaging modalities for identifying the parathyroid glands were evaluated in 177 patients whose intact parathyroid hormone (PTH) levels were <9 pg/mL after the initial PTx. Additional PTx procedures were performed on 7 of 114 patients whose intact PTH levels were >9 ng/mL after PTx, and the diagnostic validities of the imaging modalities for the remnant parathyroid glands were evaluated. A combination of CT, US, and MIBI scans achieved the highest diagnostic accuracy (75%) for locating bilateral upper and lower parathyroid glands before initial PTx. The accuracies of CT, US, and MIBI scans with respect to locating remnant parathyroid glands before additional PTx were 100%, 28.6%, and 100%, respectively. A combination of CT, US, and MIBI scans is useful for initial PTx for SHPT, and CT and MIBI scans are useful imaging modalities for additional PTx procedures.

P1590AMORPHOUS CPP AND CRYSTAL CPP OF HEMODIALYSIS PATIENTS

2020 ◽  
Vol 35 (Supplement_3) ◽  
Author(s):  
Kimihiko Nakamura ◽  
Yuki Nakayama ◽  
Toshiya Hiroyoshi ◽  
Naohito Isoyama ◽  
Kouki Fujikawa ◽  
...  
Keyword(s):  
Multivariate Analysis ◽  
Ldl Cholesterol ◽  
Gel Filtration ◽  
Intact Pth ◽  
Filtration Method ◽  
Clinical Variables ◽  
Calciprotein Particles ◽  
The Difference ◽  
Stage Renal Disease ◽  
End Stage

Abstract Background and Aims Serum calciprotein particles (CPP) are increased in CKD patients and correlated with vascular stiffness and calcification. CPPs are can be distinguished amorphous and crystal CPPs. Crystal CPPs are non-incubated CPPs and amorphous CPPs are the difference(Δ) incubated CPPs and non-incubated CPPs. In this study, we evaluated amorphous CPPs and crystal CPPs in hemodialysis (HD) patients. Method 183 end stage renal disease patients undergoing HD (57.6% men, median age 71, dialysis period; 98.4 ± 87 months) were treated in single hospital. Serum CPP levels were measured by the gel filtration method. Incubated CPPs were incubated in 24°C, 24hous then were measured similarly. We assessed the association of serum calcium (Ca), phosphorus (P), intact-PTH, FGF21, LDL-cholesterol, CRP with CPP. Results In multivariate analysis, P remained significant independent factors for the non-incubated CPP levels. Ca, P remained significant independent factors for the incubated CPP levels. P remained significant independent factors for the Δ CPP levels. The association of ΔCPP levels / the incubated CPP levels with clinical variables are examined. In multivariate analysis, hemoglobin remained significant independent factors. Conclusion Amorphous CPPs and crystal CPPs could each be a prognostic factors. Figure: Association of ΔCPP levels / incubated CPP levels with clinical variables. Univariate Multivariate

Spectrophotometric assay for urinary N-acetyl-beta-D-glucosaminidase activity.

1981 ◽  
Vol 27 (7) ◽  
pp. 1180-1185 ◽  
Author(s):  
E Horak ◽  
S M Hopfer ◽  
F W Sunderman
Keyword(s):  
Renal Insufficiency ◽  
Coefficient Of Variation ◽  
Gel Filtration ◽  
Nickel Chloride ◽  
Spectrophotometric Assay ◽  
Enzymic Hydrolysis ◽  
Experimental Induction ◽  
Hydrolysis Of ◽  
Urine Specimens ◽  
Healthy Persons

Abstract An improved assay for N-acetyl-beta-D-glucosaminidase activity in urine is described that involves (a) gel filtration to separate the enzyme from inhibitors in urine, (b) enzymic hydrolysis of p-nitrophenyl-N-acetyl-beta-D-glucosaminide at pH 4.4, and (c) spectrophotometry of the liberated p-nitrophenylate. Measurements of activity of the enzyme in 58 urine specimens correlated closely (r = 0.9998) with results by an established procedure. The within-run coefficient of variation (CV) was 3.7%; the between-run CV averaged 6.8%. Reference values for the activity were established by assays of urine specimens from 135 healthy persons, age two weeks to 52 years. Efficacy of the assay for detection of nephrotoxicity was demonstrated in rats after experimental induction of reversible renal insufficiency by intraperitoneal injection of nickel chloride. Clinical application of the assay in approximately 1000 patients corroborated its utility for detection and monitoring of renal disorders.

scholarly journals Reversible binding of actin to gelsolin and profilin in human platelet extracts.

1987 ◽  
Vol 105 (2) ◽  
pp. 833-842 ◽  
Author(s):  
S E Lind ◽  
P A Janmey ◽  
C Chaponnier ◽  
T J Herbert ◽  
T P Stossel
Keyword(s):  
Actin Filament ◽  
Human Platelet ◽  
Gel Filtration ◽  
Actin Assembly ◽  
Reversible Binding ◽  
Calcium Dependent ◽  
High Affinity ◽  
Temporally Correlated

This paper documents the reversible appearance of high-affinity complexes of profilin and gelsolin with actin in extracts of platelets undergoing activation and actin assembly. Sepharose beads coupled to either monoclonal anti-gelsolin antibodies or to polyproline were used to extract gelsolin and profilin, respectively, from EGTA-containing platelet extracts and determine the proportion of these molecules bound to actin with sufficient affinity to withstand dilution (high-affinity complexes). Resting platelets (incubated for 30 min at 37 degrees C after gel filtration) contained nearly no high-affinity actin/gelsolin or actin/profilin complexes. Thrombin, within seconds, caused quantitative conversion of platelet profilin and gelsolin to high-affinity complexes with actin, but these complexes were not present 5 min after stimulation. The calcium-dependent actin filament-severing activity of platelet extracts, a function of free gelsolin, fell in concert with the formation of EGTA-stable actin/gelsolin complexes, and rose when the adsorption experiments indicated that free gelsolin was restored. The dissociation of high-affinity complexes was temporally correlated with the accumulation of actin in the Triton-insoluble cytoskeleton.

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