A radioimmunoassay for 1,25-dihydroxycholecalciferol.

1980 ◽  
Vol 26 (5) ◽  
pp. 562-567 ◽  
Author(s):  
R Bouillon ◽  
P De Moor ◽  
E G Baggiolini ◽  
M R Uskokovic

Abstract We describe a radioimmunoassay for 1,25-dihydroxycholecalciferol in human serum. We raised antisera in rabbits to 1,25-dihydroxycholecalciferol-3-hemisuccinate coupled to bovine serum albumin, and obtained sensitive, high-titer antibodies. These antibodies had a high affinity for 1,25-dihydroxycholecalciferol and cross reacted mainly with 25-hydroxycholecalciferol and 24,25-dihydroxycholecalciferol. Addition of 1 mL of normal rabbit serum per liter reduced this interference to 5 and 4%, respectively. However, these interfering steroids are present in large excess, so extensive purification of 1,25-dihydroxycholecalciferol from serum is necessary. The steroid was extracted with ethyl acetate/cyclohexane, purified on Sephadex LH-20, and then chromatographed on a column of silicic acid. The radioimmunoassay is sensitive to 5 pg/tube (3 ng/L of serum). The between-assay CV was 14%. The mean concentration of 1,25-dihydroxycholecalciferol in the serum of 54 healthy adults was 38 (SD 12) ng/L, with no sex-related difference. The assay was further validated by the finding of low or undetectable concentrations in patients with chronic renal failure and of increased concentrations in the serum of patients with primary hyperparathyroidism. In comparison with previously described methods, the major advantage of the present assay is the use of stable gamma-globulins, which are available in large amounts, as binding protein.

1980 ◽  
Vol 26 (5) ◽  
pp. 562-567 ◽  
Author(s):  
R Bouillon ◽  
P De Moor ◽  
E G Baggiolini ◽  
M R Uskokovic

Abstract We describe a radioimmunoassay for 1,25-dihydroxycholecalciferol in human serum. We raised antisera in rabbits to 1,25-dihydroxycholecalciferol-3-hemisuccinate coupled to bovine serum albumin, and obtained sensitive, high-titer antibodies. These antibodies had a high affinity for 1,25-dihydroxycholecalciferol and cross reacted mainly with 25-hydroxycholecalciferol and 24,25-dihydroxycholecalciferol. Addition of 1 mL of normal rabbit serum per liter reduced this interference to 5 and 4%, respectively. However, these interfering steroids are present in large excess, so extensive purification of 1,25-dihydroxycholecalciferol from serum is necessary. The steroid was extracted with ethyl acetate/cyclohexane, purified on Sephadex LH-20, and then chromatographed on a column of silicic acid. The radioimmunoassay is sensitive to 5 pg/tube (3 ng/L of serum). The between-assay CV was 14%. The mean concentration of 1,25-dihydroxycholecalciferol in the serum of 54 healthy adults was 38 (SD 12) ng/L, with no sex-related difference. The assay was further validated by the finding of low or undetectable concentrations in patients with chronic renal failure and of increased concentrations in the serum of patients with primary hyperparathyroidism. In comparison with previously described methods, the major advantage of the present assay is the use of stable gamma-globulins, which are available in large amounts, as binding protein.


1987 ◽  
Vol 33 (8) ◽  
pp. 1382-1386 ◽  
Author(s):  
K Liewendahl ◽  
S Tikanoja ◽  
H Mähönen ◽  
T Helenius ◽  
M Välimäki ◽  
...  

Abstract The mean concentration of free thyroxin (FT4) in serum, as determined by direct equilibrium dialysis, was decreased in patients with chronic renal failure (CRF) and increased in patients with various other nonthyroidal illnesses (NTI). The mean concentration of dialyzable free triiodothyronine (FT3) in serum was equally low in both groups of patients. Patients with CRF of various etiology but a similar degree of renal failure as estimated from serum creatinine assay had very similar concentrations of FT4 and FT3 in their serum. Mean thyroxin (T4) and triiodothyronine (T3) concentrations in serum were decreased in CRF and NTI, whereas the mean reverse-T3 concentration in serum was normal in CRF and increased in NTI. T4-binding globulin and albumin were markedly decreased in CRF and NTI; T4-binding prealbumin was increased in CRF and decreased in NTI. The mean concentration of nonesterified free fatty acids (FFA) in serum was increased in NTI but not in CRF. The weak, but significant, positive correlation observed between FT4 and FFA in serum (r = 0.34, P less than 0.01) in NTI indicates that the increase in serum FT4 in this group of patients could be an effect, at least in part, of FFA competing with T4 for binding sites on serum proteins. The stronger correlation detected between the serum FT4 concentration and the FFA/albumin molar ratio in serum (r = 0.60, P less than 0.001) demonstrates the importance of a low albumin concentration for expression of the effect of FFA on FT4 in severe systemic illnesses.


2017 ◽  
pp. 88-92
Author(s):  
Van Hien Pham ◽  
Huu Vu Quang Nguyen ◽  
Tam Vo

Background: Cardiovascular diseases are the leading cause of death in patients with chronic renal failure. When a patient undergoes dialysis, making AVF or AVG causes cardiovascular events. Understanding the relationship between complications: hypertension, heart failure, AVF or AVG (formation time, position, diameter) helps us monitor, detect, prevent and treatment of complications to limit the risk of death in patients with dialysis. Objective: Relationship between cardiovascular diseases and anatomosis of arteriovenous fistular in patients with regularly hemodialysis at Cho Ray Hospital. Methods: A cross-sectional study was conducted at Cho Ray Hospital from 2015 to 2016. The survey some cardiovascular diseases are done by clinical examination, tests for diagnostic imaging such as X-ray, electrocardiogram and echocardiogram: heart and diameter of anastomosis AVF, AVG. Results: The study population included 303 patients with chronic renal failure who were dialysis. Of which, patients aged 25-45 accounted for the highest proportion (43.9%). The proportion of male and female patients was similar (48.5% and 51.5% respectively). The mean value of systolic blood pressure on patients made AVF, AVG less than 12 months is higher than patients made AVF, AVG over 12 months, and there is negative correlation (r = -0.43) between AVF, AVG and systolic blood pressure (p <0.05). The mean value of diastolic blood pressure on patients made AVF, AVG less than 12 months is lower than patients made AVF, AVG over 12 months, and and there is positive correlation (r = -0.43) between AVF, AVG and diastolic blood pressure (p <0.05) (p <0.05). The prevalence of patients with heart failure made AVF, AVG over 12 months is higher than that of the under 12 months group, there is a negative correlation (r = - 0.43) between AVF, AVG diameter and EF index (p <0.05). Conclusion: It is important to note the diameter of anastomosis AVF, AVG in patients with chronic renal failure dialysis to limit cardiovascular complications, especially heart failure. Key words: Chronic kidney disease, hemodialysis.


2017 ◽  
Vol 68 (6) ◽  
pp. 1325-1328
Author(s):  
Andrada Raluca Doscas ◽  
Mihail Balan ◽  
Mihai Liviu Ciofu ◽  
Doriana Agop Forna ◽  
Marius Cristian Martu ◽  
...  

Chronic kidney disease (CKD) is a multifactorial syndrome and a global health concern. As renal function declines, there is a progressive deterioration of mineral homeostasis. Starting from stage 3 of CKD oral manifestations of mineral disorders can occasionally appear and become more frequent and evident in stage 5. We retrospectively analysed 43 patients diagnosed with end stage renal failure undergoing dialysis, hospitalized in our clinic for different oral and maxillofacial pathologies. The mean dialysis period was 5.43 years. Radiographic alterations afecting the jaws were found in all patients. The most common feature was partial or total loss of lamina dura, followed by alterations of the bony trabeculae. 9 patients presented brown tumors which are considered the final stage of secondary hyperparathyroidism associated with renal failure.


1994 ◽  
Vol 87 (3) ◽  
pp. 363-368 ◽  
Author(s):  
Aubrey Blumsohn ◽  
Brian Morris ◽  
Richard Eastell

1. Stable strontium (Sr) has been proposed as an alternative to calcium (Ca) isotopes for the measurement of intestinal Ca absorption. The aim of this study was to compare the time course and fractional absorption of Ca and Sr, when both are measured using dual-tracer techniques. 2. 45Ca and Sr absorption tests were carried out on consecutive days in patients with osteoporosis (n = 10) or chronic renal failure (n = 7). Both tests were repeated in four patients with chronic renal failure after treatment with calcitriol (1 μg daily for 10 days). 3. The time course of Ca absorption was determined using the 85Sr (intravenous)/45Ca (oral) dual-tracer technique, and the time course of Sr absorption using 85Sr (intravenous)/stable Sr (oral). Oral tracers were administered on consecutive days with a test meal containing 5.3 mmol of Ca and 2.5 mmol of either stable Sr or Ca carrier. The fractional absorption of 45Ca and Sr at 6 h (FA360) and the absorption rate as a function of time were calculated by deconvolution. 4. The mean FA360 for Sr (20.2%) was lower than the mean FA360 for 45Ca (37.8%, P < 0.001, paired t-test), but the time course of Sr absorption was similar to that of Ca. There was a significant correlation between the FA360 for 45Ca and Sr, although the relationship was improved by including a quadratic term (R2 = 0.89, P < 0.001, significance of quadratic term, P < 0.05). After 1,25-dihydroxyvitamin D treatment, the FA360 of stable Sr increased 4.29-fold, whereas the FA360 of 45Ca increased only 2.4-fold. 5. Although the fractional absorption of Sr determined by dual-tracer deconvolution was the best predictor of FA360 for 45Ca, little was lost by confining the analysis to a single serum Sr measurement taken 3 h or more after oral administration. 6. We conclude that Sr absorption is qualitatively similar to that of Ca, although absorption of Sr is much lower than that of Ca. Furthermore, the relationship does not appear to be linear. Stable Sr may be useful in place of Ca isotopes in the routine clinical evaluation of Ca absorption.


1981 ◽  
Author(s):  
S A Burstein ◽  
S K Erb ◽  
J W Adamson ◽  
L A Harker

Previous studies from our laboratory have suggested that the numbers of CFU-M do not increase primarily in response to acute thrombocytopenia. To determine the effect and specificity of prolonged thrombocytopenia on CFU-M number, mice were given 4 intravenous injections on alternate days of multiply absorbed rabbit anti-mouse platelet serum (APS), while control animals received a similar regimen of rabbit anti-mouse red cell serum (ARS), normal rabbit serum (NRS), or phosphate-buffered saline (PBS). Two days aftefcthe final injection, the mean platelet count was.0.314 ± 0.129 × 106/ul in animals given APS vs. 1.105 ± 0.048 × 106/ul in animals given other regimens. The numbers of CFU-M, day 7 and day 14 erythroid burst forming cells (BFU-E), and granulocyte-macrophage colony forming cells (CFU-C) were determined in humerus and spleen.The generalized increase in progenitor cells in marrow in response to APS together with increases in CFU-M in spleen following ARS and NRS indicate that these cells may respond nonspecifically to foreign protein. The data suggest that the elevation in CFU-M numbers with chronic immune thrombocytopenia is at least partially independent of the platelet count.


1985 ◽  
Vol 31 (11) ◽  
pp. 1815-1819 ◽  
Author(s):  
B W Hollis ◽  
J L Napoli

Abstract We describe a faster, more-sensitive radioimmunoassay for vitamin D in plasma. Antibodies were generated in rabbits immunized with a new vitamin D analog, the 23,24,25,26,27-pentanor-C(22)-carboxylic acid of vitamin D, coupled directly with bovine serum albumin. After several months, Rivanol-treated sera from the rabbits contained high-titer antibodies, as determined by their abilities to bind 25-hydroxy-[3H]cholecalciferol. The antibody, used at a 1:15 000 final dilution, cross reacted equally with all cholecalciferol and ergocalciferol metabolites tested except 1,25-dihydroxycalciferol metabolites and the parent calciferols. 25-Hydroxycalciferol and similar forms were efficiently extracted from plasma or serum with acetonitrile. We separated bound from free 25-hydroxy-[3H]cholecalciferol by using a second antibody: goat antiserum to rabbit serum. The detection limit of the assay was 3.0 pg of 25-hydroxycholecalciferol and its equivalents per tube; thus only 1 microL of plasma is needed per assay tube. Results compared well with those from a liquid-chromatographic procedure involving specific ultraviolet detection of 25-hydroxycalciferol in plasma.


1975 ◽  
Vol 21 (4) ◽  
pp. 521-527 ◽  
Author(s):  
Noritake Asato ◽  
Maria van Soestbergen ◽  
F William Sunderman

Abstract Binding of 63Ni(Il) to ultrafiltrable constituents of rabbit serum was studied (a) after in vitro incubation (2 h, 37 °C) of rabbit serum with 63NiCl2 (10-100 µmol/liter), and (b) at intervals (0.25-2 h) after in vivo administration of 63NiCl2 (40-160 µmol/kg body wt, i.v.). Serum ultrafiltrates were fractionated by thin-layer chromatography, and the separated compounds made visible by autoradiography and by ninhydrin staining. Several (≃5) ultrafiltrable 63Ni-complexes were demonstrable as distinct radiodense 63Ni-bands with chromatographic mobilities corresponding to those of ninhydrin-positive bands. Unbound 63Ni(II) was not detected in serum ultrafiltrates in either the in vitro or in vivo experiments. In sera (n = 10) incubated in vitro with 63Ni(II) (10 µmol/ liter), the mean percentage of ultrafiltrable 63Ni was 36% (range = 33-38) of total serum 63Ni. In contrast, in sera (n = 10) obtained 2 h after i.v. injection of 63Ni(II) (40 µmol/kg), the mean concentration of total serum 63Ni was 10.8 µmol/liter (range = 6-14), and the mean percentage of ultrafiltrable 63Ni was 15% (range = 9-21) of total serum 63Ni. The disparity between the percentages of ultrafiltrable 63Ni obtained in vitro and in vivo was obviated when the in vivo experiments were performed in rabbits bilaterally nephrectomized, with ligated common bile ducts. This investigation confirms the existence of several nickel receptors in serum ultrafiltrates and substantiates the role of ultrafiltrable complexes in the excretion of nickel.


1986 ◽  
Vol 6 (3) ◽  
pp. 130-135 ◽  
Author(s):  
Kostas Sombolos ◽  
Peter McNamee ◽  
Ahmed Mitwalli ◽  
Sol Rabinovich ◽  
Dimitrios G. Oreopoulos

From October 1977 to October 1985, our pathology department did autopsies on 19 patients (14 men, five women) treated by CAPD for four to 55 (mean 29.3) months. Their mean age was 60.2 (range 28–79) years and the primary renal diseases were diabetes mellitus (eight), nephrosclerosis (five), polycystic kidneys (three), chronic glomerulonephritis (one) and chronic renal failure associated with sarcoidosis and congestive cardiomyopathy in two. During the same period, the authors selected as controls 18 autopsied patients (14 men, four women), who had not had chronic renal failure, and these were matched with the CAPD patients for age, sex, longstanding hypertension and insulin-dependent diabetes. Direct causes of death for CAPD patients were cardiovascular incidents (12) infection (5), pancreatitis (1) and lung cancer (1); in controls, the causes were cardiovascular in 11 and infection in two. Thirteen of the CAPD and 12 autopsied controls had coronary artery stenosis equal to or greater than 70%, and affecting one or more arteries. The mean weight of organs in CAPD patients and controls were similar except for kidneys and the spleen; we found the latter weighed more in those on CAPD (p = 0.002). In CAPD patients the most important organ changes were: evidence of myocardial infarction, old or acute, in nine, acquired cystic disease of the kidney in five, and thickening and adhesions of peritoneum in nine and five respectively.


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