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Albumin (Alb) and transferrin (Tf) polymorphism in plasma of Callithricidae was investigated by means of starch gel electrophoresis. In 52 blood samples of three species (Saguinus mystax, S. oedipus and S. labiatus), four Alb phenotypes (Alb 1, Alb 2, Alb 3 and Alb 2-3) and two Tf phenotypes (Tf 1 and Tf 2) were observed. No Alb variant was found in S. oedipus and S. mystax.
BACKGROUND: Hemodialysis (HD) is often accompanied by adverse effects, such as tissue ischemia. We have already observed an increase in plasma adenosine (ADO) levels during HD sessions, which may be the result of tissue ischemia. Here we evaluate the influence of the dialysis membrane on two sensitive and early markers of ischemia: ADO and ischemia-modified albumin (IMA). METHODS: We included in the study 50 patients with end-stage renal failure, 39 hemodialyzed (mean age 61+/-24 years; 24 male; membranes: 23 synthetic and 16 cellulose based) and 11 undialyzed (mean age 55+/-12 years; 6 male), and 10 healthy subjects (mean age 47+/-11 years; 4 male). We compensated for hemoconcentration during HD by measuring either the IMA to albumin (Alb) or the ADO to Alb ratio. RESULTS: Under basal conditions, the IMA to Alb ratio was not significantly different in patients and controls and HD did not significantly modify this ratio. Conversely, the ADO to Alb ratio (mean+/-SD in micromol/g) was higher in patients before HD compared with either undialyzed patients or controls (before HD: 0.077+/-0.02; undialyzed patients: 0.026+/-0.11; controls: 0.022+/-0.01). During HD, there was a significant increase in the ADO to Alb ratio (before HD: 0.077+/-0.02; after HD: 0.09+/-0.029; p<.01). We found no significant difference in the IMA to Alb or ADO to Alb ratio using either synthetic or cellulose-based membranes. CONCLUSIONS: We concluded that ADO is a more sensitive marker of ischemia than IMA and that, under our HD conditions, the ischemia caused by HD was very weak, independent of the dialysis membrane.
The role of plasma proteins on the cellular uptake of lipophilic substrates has perplexed investigators for many years. We tested the hypothesis that an ionic interaction between the protein-ligand complex and hepatocyte surface may be responsible for supplying more ligand to the cell for uptake. The surface-charged groups on albumin were modified to yield proteins having a range of isoelectric points (ALB, ALBs, ALBm, ALBe had values of 4.8-5.0, 4.5-4.7, 3.0-3.5, 8.4-8.6, respectively). [3H]-Palmitate uptake studies were performed with adult rat hepatocyte suspensions using similar unbound ligand fractions in the presence of the different binding proteins. Mass spectrometry, isoelectric focusing (pI), and heptane:water partitioning were used to determine protein molecular weight, pI, and protein-palmitate equilibrium binding constant, respectively. Hepatocyte [3H]-palmitate clearance in the presence of ALBs and ALBm were significantly lower (p < 0.05) than ALB, whereas [3H]-palmitate clearance in the presence of ALBe was significantly higher (p < 0.05) than ALB. The data were consistent with the notion that ionic interactions between extracellular protein-ligand complexes and the hepatocyte surface facilitate the uptake of long-chain fatty acids.
Nitric oxide (NO) is an endogenous vasodilator and modulator of inflammation. During endotoxemia, the beneficial effects of NO are overwhelmed by the inflammatory cascade, resulting in a functional depletion of NO. S-nitroso-albumin (S-NO-alb) exists as a novel and highly stable NO thiol complex that slowly releases NO into the vascular micro-environment. Using a porcine model, we examined the ability of intravenous S-NO-alb to modulate cardiopulmonary dysfunction characteristic of endotoxemia. Pigs were anesthetized, instrumented for standard cardiopulmonary function measurements, and randomly assigned to receive: (i) albumin + saline; (ii) albumin + LPS; or (iii) S-NO-alb + LPS. Cardiopulmonary parameters were evaluated every 30 min and ex vivo phorbol myristate acetate (PMA)-stimulated superoxide release was serially determined as a marker of in vivo neutrophil priming. Lung myeloperoxidase (MPO) activity was measured as a marker of neutrophil migration into the lung. LPS-induced cardiopulmonary dysfunction was characterized by a sustained elevation in mean pulmonary arterial pressure, pulmonary vascular resistance, and peak intratracheal pressure, as well as a reduction in cardiac index, stroke volume index and PaO(2) over 6 h. Pretreatment with S-NO-alb attenuated LPS-induced cardiopulmonary dysfunction without adversely affecting systemic hemodynamics. Moreover, S-NO-alb blunted the LPS-induced hypoxemic response and reduced neutrophil activation. S-NO-alb did not, however, attenuate LPS-induced increases in lung MPO. Our results suggest that S-NO-alb can selectively modulate endotoxin-induced pulmonary dysfunction, attenuate neutrophil priming and block the early mortality (40%) in this model.
Increasing, the colloid osmotic pressure (COP) of blood cardioplegia (BCP) may reduce myocardial edema and preserve cardiac function following cardiopulmonary bypass (CPB). The purpose of this study was to quantify the effects of albumin (ALB) supplementation on cardioplegia COP through an in vitro analysis. A self-contained cardioplegia delivery system administered supplemental ALB to four BCP ratios (1:1, 4:1, 8:1, and 20:1). In Group A, 25% ALB was combined with BCP at four delivery rates (0, 13, 25, and 50 mL ALB/L BCP), with a delivery rate of 0 mL ALB/L BCP serving as the control for all groups. Twenty-five percent ALB was added to crystalloid to create carrier solutions containing 12.5, 25, or 50 g ALB/L in Group B, while Group C combined an ALB delivery rate of 50 mL ALB/L BCP with each of the three carrier solutions. End-points included initial and post-supplementation hematocrit, total serum protein (TSP), and COP. Without supplemental ALB, TSP was less affected with increasing blood to crystalloid ratios (1:1-81.7 +/- 6.2%, 4:1-40.6 +/- 5.1%, 8:1-20.6 +/- 4.1%, 20:1-6.0 +/- 5.7%). The TSP of 1:1 and 4:1 BCP increased (p < .0003 and p < .02) across all methods of supplementation, while 8:1 BCP was similarly increased (p < .008), except with 12.5 and 25 g ALB/L carrier solutions. The greatest change from baseline COP was seen with the lower blood to crystalloid ratios (1:1-64.3 +/- 5.0% and 4:1-39.5 +/- 10.5%). In higher ratios, the effects of dilution were less profound (14.6 +/- 4.2 +/- 4.2% and 20:1-6.0 +/- 1.9%). COP of 1:1 BCP increased (p < .008) whenever ALB was added. In conclusion, TSP and COP of blood cardioplegic solutions is increased by supplemental albumin administration with quantitative enhancement dependent upon the dilutional effects of the blood to crystalloid ratio.
OBJECTIVE: To study albumin adduct with naphthalene metabolites, namely 1,2-naphthoquinone (1,2-NPQ) and 1,4-naphthoquinone (1,4-NPQ), as a potential biomarker for intermediate/long-term exposure to polycyclic aromatic hydrocarbons (PAH) in coke oven workers. METHODS: Twenty-eight coke oven workers and 22 control workers were recruited from a cokery. Spot urine and venous blood samples were collected from the workers after four continuously working days and personal information was obtained by questionnaire. Plasma albumin adduct was detected with gas chromatography-mass spectrometry. RESULTS: Albumin adduct with 1,2- & 1,4-NPQ (1,2-NPQ and 1,4-NPQ), respectively, were detected in all coke oven workers and controls. Median plasma level of 1,2-NPQ-Alb in coke oven workers was significantly higher than that in controls (76.6 pmol/g vs. 44.9 pmol/g, P < 0.01). However, there was no significant difference in plasma median level of 1,4-NPQ-Alb between the two groups (48.6 pmol/g vs. 44.2 pmol/g, P > 0.05). Plasma level of 1,2-NPQ-Alb was significantly higher than that of 1,4-NPQ-Alb in coke oven workers. Urine levels of naphthalene, 1-naphthol, 2-naphthol and 1-pyrenol in coke oven workers correlated significantly with their plasma level of 1,2-NPQ-Alb (Pearson coefficient of correlation greater than 0.371, P < 0.01), but did not do significantly with 1,4-NPQ-Alb. CONCLUSION: Plasma level of 1,2-NPQ-Alb could effectively reflect their magnitude of personal internal dose of exposure to air PAH, so it could be used as a potential biomarker to evaluate their intermediate/long-term exposure to PAH in coke oven workers.
OBJECTIVE: To explore whether bone marrow stem cells (MSCs) from adult mice can be induced to differentiate into hepatocytes by hepatocyte growth factor (HGF) alone and the time phase characteristics in the differentiation progress. METHODS: Adult mouse MSCs were treated with or without 100 ng/ml HGF, on days 0, 7, 14, 21, and 28. The morphologic characteristics of the cells were examined; the albumin (ALB), AFP mRNA was analyzed sub-quantively using reverse transcription polymerase chain reaction (RT-PCR) and immumohistochemistry techniques. The expression of ALB, AFP and CK19 were detected by using anti-ALB, AFP and CK19 antibodies. RESULTS: Freshly isolated adult mouse MSCs expressed ALB and AFP mRNA weakly; in the group without HGF, no ALB mRNA was detected on day 7. The expression of AFP mRNA was reduced significantly on day 7, and could not be detected anymore after day 14. In the HGF treated group, ALB mRNA was not detected on day 7, but the positive lane appeared again on day 14, and the expression of ALB mRNA was increased on day 21 but reduced in the following days. The AFP mRNA was positive at all times, however it tended to decrease after day 14 in the HGF treated groups. The result of immumohistochemistry was consistent with that of RT-PCR, and CK19 was always negative. CONCLUSION: Adult mouse MSCs can be induced into hepatocyte differentiation in vitro. The optimal time for the induction was 2 to 3 weeks.
We thought that nutritional parameters in laboratory data might be able to express quality of life (QOL). Therefore, in 70 patients with malignant chest diseases (NSCLC, 42 patients; SCLC, 15; lung metastasis, 7; others, 6), the correlation between nutritional parameters of total protein (Tp), serum albumin (Alb), and serum (cholinesterase (ChE] and Karnofsky Performance Status scale (KPS) was investigated. Then, in 24 patients with them (NSCLC, 12; SCLC, 6; lung metastasis, 4; others 2), Alb and ChE were compared to the EORTC Core Quality of Life Questionnaire and Lung Cancer-Specific Questionnaire Module (QS). Results were as follows: 1) KPS and nutritional parameters correlated (Tp. r = 0.55, p less than 0.001; Alb, r = 0.60, p less than 0.001, ChE, r = 0.60; p less than 0.001). 2) The cores for Functional Status (FS) and Disease and Treatment-related symptoms (Sym) in QS and parameters of Alb and ChE correlate (FS v.s. Alb, p less than 0.01; Sym v.s. Alb, p less than 0.01; FS v.s. ChE, p less than 0.05; and Sym v.s. ChE, p less than 0.05). Moreover, the scores of Psychological Distress in QS and Alb showed a correlation (p less than 0.05). It is considered that nutrition and part of QOL (KPS and FS + Sym in QS, that is to say, "objective" functional activity and "subjective" functional activity and symptoms) correlate, and that nutritional parameters are useful to evaluate QOL.
Possible effects of single point mutations on the ligand-binding capabilities of human serum albumin (Alb) were investigated by studying the interactions between the strongly bound drugs warfarin, salicylate, and diazepam and five structurally characterized genetic variants of the protein. Equilibrium dialysis data, obtained with the variants and normal serum Alb, revealed pronounced reductions in high affinity binding of all three ligands to Alb Canterbury (313 Lys----Asn) and to Alb Parklands (365 Asp----His). By contrast, unchanged binding of the drugs was found in the case of Alb Verona (570 Glu----Lys). Different effects on binding were observed for the other two variants. Salicylate was the only drug bound with a lower affinity to Alb Niigata (269 Asp----Gly), whereas binding of both salicylate and diazepam to Alb Roma (321 Glu----Lys) were moderately reduced. In about half of the cases of diminished binding, the primary association constant was reduced by 1 order of magnitude, giving rise to an increase in the unbound fraction of the drugs of 500% or more at therapeutically relevant molar ratios of drug and protein. Changes in protein charge seem to be of only minor importance for reduced binding. More likely, conformational changes in the 313-365 region of the proteins are the main cause for diminished binding of these diverse ligands, which probably have different high affinity binding sites. The specific reduction in salicylate binding after modification of residue 269 may be due to conformational changes at or close to the salicylate binding site.
In an attempt to define the nature of renal selectivity in diabetes mellitus, we have determined the free sulfhydryl (SH) groups of serum and urinary albumin in 9 normal subjects and 24 diabetic patients with various grades of renal involvement, as defined by their urinary excretion rates of albumin (alb. UER): 8 with alb. UER less than 10 micrograms/min (Group A), 6 with alb. UER between 10 and 30 micrograms/min (Group B), 5 with alb. UER between 30 and 200 micrograms/min (Group C) and 5 with alb. UER greater than 200 micrograms/min (Group D). The free SH group content of urinary albumin was three to four fold increased in comparison with its serum homologue in normal subjects and in diabetics with normal or slightly increased alb. UERs (Groups A and B). Diabetics in Group C showed a two-fold increase in free SH groups of urinary albumin compared to serum albumin and diabetics with clinical nephropathy (Group D) showed no increase at all. The SH group content of urinary albumin correlated in all diabetics with the concentration of glycosyl albumin and the of urinary/serum albumin SH groups ratio was inversely correlated with alb. UER. From these observations concerning the selectivity properties of the renal filter in normal and diabetic subjects, it is concluded that the mechanism for progression of diabetic nephropathy may be the hyperfiltration of albumin with an altered conformational state.
We studied the filtration characteristics of glomeruli isolated from the superficial and deep renal cortex of Munich-Wistar rats (180 to 300 gm) to determine whether the ultrafiltration coefficient (Kf) or hydraulic conductivity (Lp) of the glomerular capillary is altered during expansion of extracellular fluid volume with isotonic protein-free (SAL) or hyperoncotic (ALB) solutions. SAL rats were given an initial infusion of 1% dextrose in lactated Ringer's solution (0.1 ml/gm body weight) followed by a maintenance infusion for 18 to 20 hours. ALB rats were given hyperoncotic bovine serum albumin (0.01 gm/gm body weight) intravenously 2 hours before sacrifice. At sacrifice, urine flow rate and fractional excretion of sodium were increased and plasma renin activity decreased in both SAL and ALB rats. Expansion of intravascular volume was evidenced by a decrease in the hematocrit of ALB but not of SAL rats; plasma protein concentration was increased in ALB and unchanged in SAL rats. Kf and Lp were estimated during in vitro filtration. Because SAL rats were significantly larger than ALB rats, results were compared with those in separate size-matched controls. Kf of superficial glomeruli of large control and SAL rats did not differ (3.7 +/- 0.4 and 3.5 +/- 0.4 nl/min/mm Hg, respectively). Lp, of the same glomeruli, calculated as the quotient of Kf and estimated filtering area (LpD), were also comparable (1.9 +/- 0.2 and 1.8 +/- 0.2 microliter/min/mm Hg/cm2). Kf of superficial glomeruli of ALB rats was significantly greater than Kf of size-matched control rats (3.7 +/- 0.3 vs. 2.9 +/- 0.1 nl/min/mm Hg, P less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)
Packed cell volume and plasma total protein (TP), serum albumin (Alb) and globulin (Glb), and plasma ionized calcium (PCa) concentrations, blood viscosity (BV), and plasma viscosity (PV) were measured in 42 horses at rest and after the cross country jumping phase of a horse trial competition. The BV and PV were determined at 6 shear rates (230, 115, 46, 23, 11.5, 5.75 s-1), using a digital rotational cone and plate microviscometer. A paired t-test was used to determine differences between PCV, TP, Alb, Glb and PCa values at rest and after exercise. The PCV, TP, Alb, and Glb values increased (P < 0.05) in horses after exercise. The PCa concentration decreased (P < 0.05) in horses after exercise. Mean BV and PV in the 42 horses at rest and after exercise were fitted to an asymptotic function. Significant (P < 0.05) correlation at all shear rates was seen between BV at rest and PCV, TP, Alb, Glb, and PCa values at rest; and between BV after exercise and PCV, TP, Alb, Glb, and PCa values after exercise. Significant correlation was not seen between PV at rest and TP, Alb, Glb, and PCa at rest, or between PV after exercise and TP, Alb, Glb, and PCa concentrations after exercise at any shear rate.
One possible cause of pulmonary failure after trauma is the type of asanguinous fluid used for resuscitation, i.e., crystalloid or colloid. To investigate this issue, patients having a laparotomy for trauma randomly received either Ringer's lactate (RLS) alone or Ringer's lactate with albumin (ALB). Both groups received washed red cells. Test fluids and red cells were given before and during operation to restore vital signs and hematocrit to normal. Pulmonary function tests were performed for 5 days after operation. One hundred and forty-one cases were studied (84 RLS, 57 ALB). The volume of asanguinous resuscitation fluid infused was 5.37 +/- 3.38 liters (-x +/- SD) for RLS and 5.87 +/- 3.05 liters for ALB. ALB cases received 213 +/- 130 gm of albumin. Red cell transfusions were 1.5 +/- 2.9 units for RLS and 2.0 +/- 3.1 for ALB. Six patients died (three RLS, there ALB). Three RLS and six ALB patients received intermittent mandatory ventilation or continous positive air pressure after operation. Pulmonary function results were not significantly different between the two groups for any parameter on any day after operation. Results of a randomized trial in human subjects of resuscitation with crystalloid and colloid solutions for acute trauma requiring laparotomy did not reveal significant differences in (1) survival rate, (2) incidence of pulmonary failure, or (3) postoperative pulmonary function.
Beta-trace protein concentrations in cerebrospinal fluid (CSF) and serum from 113 patients with various neurological diseases and 65 controls were determined with a sensitive and specific immunonephelometric assay. In adult control patients, beta-trace concentrations were 14.6+/-4.6 mg/L in CSF and 0.46+/-0.13 mg/L in serum, that is, 32-fold higher in CSF. beta-trace levels in CSF correlated with age as well as with the albumin CSF/serum ratio (Q(Alb)), which is considered a measure for blood-CSF barrier function. The relationship between CSF beta-trace levels and elevated Q(Alb) values was studied in various neurological diseases with CSF protein increase. In spinal canal stenosis, CSF beta-trace (mean=29.5+/-10.5 mg/L) correlated positively with increasing Q(Alb) values. In bacterial meningitis, CSF beta-trace (mean=8.7+/-3.9 mg/L) remained invariant to changes of Q(Alb) values. In Guillain-Barré syndrome, CSF beta-trace (mean=14.4+/-6.8 mg/L) was below the Q(Alb)-dependent reference range. In multiple sclerosis and viral meningoencephalitis, beta-trace levels were within the reference range. Beta-trace concentration in CSF can be used in conjunction with Q(AlB) to distinguish between different neurological pathologies associated with CSF protein increase.
The effect of uremia on the binding of diazoxide to plasma proteins was studied. An equilibrium dialysis technique, using diazoxide-minus14C at approximately 30 and 300 mug/ml in the plasma phase, was used to measure diazoxide binding to plasma. Serum albumin concentration (Alb) and serum creatinine (Cr) or blood urea nitrogen (BUN) were negatively correlated. By single regression analysis, per cent free diazoxide (%FD) correlated negatively with Alb and positively with Cr or BUN. When %FD was regressed simultaneously against Alb and Cr or BUN, Alb emerged as the sole determinant of %FD (p less than 0.001), indicating that creatinine or BUN correlated with %FD by their inverse correlation to Alb rather than by an effect on drug protein binding. At the levels of Alb studied, %FD varied over a 2-fold range. In a retrospective study of the influence of uremia on diazoxide effect in hypertensive patients, a relatively low correlation (r, 0.59) was found between BUN and hypotensive effect. Prospective studies involving correlations of drug effect with renal function and %FD are required to assess the clinical importance of decreased binding of diazoxide to uremic plasma.
A macromolecular MR contrast medium (MMCM) designed to permit histochemical staining and specific tissue localization, albumin-(biotin)10-(Gd-DTPA)25 (Bio-Alb-Gd), was used in a rat model of reperfused myocardial infarction to confirm the presence and distribution of microvascular hyperpermeability. T1-weighted spin-echo images were acquired before and after administration of Bio-Alb-Gd. An avidin-biotin-complex (ABC) stain, specific for the biotinylated MR contrast medium, was used to define the MMCM distribution and to detect any regional change in microvascular permeability related to infarction. Immediately after Bio-Alb-Gd administration, the infarcted region was enhanced, with greatest signal intensity noted at the rim and less at the center. There was a gradual increase in signal intensity of the initially hypointense central region. The steady increase in signal intensity of the central region suggested convection transport of MMCM through the interstitial space and its influx into cellular compartment after leakage from the vascular compartment. Histologic findings confirmed regional microvascular hyperpermeability corresponding to the site of infarction and a predominant rim distribution of the MMCM. Bio-Alb-Gd was identified at high microscopic power in the intravascular, interstitial, and intracellular spaces at the periphery of reperfused infarcted myocardium. Bio-Alb-Gd can be used as an MR contrast medium in reperfused infarcted myocardium to confirm the existence and to localize altered microvascular permeability to macromolecules. Bio-Alb-Gd contrast technique removes all the ambiguity between the distribution of the MR or other imaging contrast agent and the distribution of the substrate for histochemical staining.
The present study was undertaken among 260 subjects [133 controls and 127 persons exposed to polycyclic aromatic hydrocarbons (PAHs) from two steel foundries and a graphite electrode producing plant in order to investigate the relationship between the benzo[a]-pyrene-diol epoxide adducts to albumin (BPDE-alb) and the intensity of exposure to PAHs. Blood samples were collected from each subject and BPDE-alb adduct determination was performed using a high-pressure liquid chromatography technique with fluorescence detection. Exposure to PAHs was assessed by measuring airborne concentration of 13 PAHs including benzo[a]pyrene (BaP) using personal air sampling and 1-hydroxypyrene excretion (1-HOP) in postshift urine. Significantly higher BPDE-alb adduct levels were observed in exposed workers compared to controls but wide interindividual variation was observed between subjects with the same level of exposure. BPDE-alb adduct level was weakly but significantly associated with the airborne concentrations of total PAHs (r = 0.35, P = 0.0001) and BaP (r = 0.30, P = 0.0001), and urinary 1-HOP excretion (r = 0.29, P = 0.0001). Alcohol and dietary habits, place of residence, and renal and hepatic status were not found to influence the concentration of BPDE-alb adducts significantly. However, for the same level of exposure to BaP, smokers had a higher probability of having an elevated BPDE-alb adduct level than non-smokers.(ABSTRACT TRUNCATED AT 250 WORDS)
The binding characteristics of [14C]ortho-toluidine (OT), a suspect human carcinogen, were investigated in male Sprague-Dawley rats. Rats were administered [14C]OT i.p. at 10, 20, 40, 50, or 100 mg/kg body weight, then sacrificed at 2, 4, 8, 18, 24, 48, or 72 h, or 7, 14, or 28 days. Hemoglobin (Hb) and albumin (Alb) were isolated from blood, and OT binding was determined by liquid scintillation counting. For Alb, peak binding occurred at 50 mg/kg at the 4-h time point (15.6 ng OT/mg Alb), while for Hb peak binding was observed at 24 h at the 100 mg/kg dose (23.0 +/- 5.1 ng OT/mg Hb). OT-Alb binding was not linear; however, OT-Hb binding appeared to increase linearly in a dose-dependent manner. Biological half-lives of OT bound to Alb or Hb were observed to be 2.6 and 12.3 days, respectively, after rats were administered a single dose of [14C]OT and sacrificed after 4 h to 28 days. The effect of route of administration on OT-Hb adduct formation was investigated, and approximately a two-fold increase in radioactivity bound to Hb was observed after i.p. administration of 100 mg/kg [14C]OT versus oral intubation. Additional studies were carried out to investigate the effect of microsomal enzyme induction. An increase in OT-Hb binding was seen in rats pretreated with phenobarbital compared to rats pretreated with beta-naphthoflavone or without pretreatment; however, this increase was not statistically significant. These results suggest that OT-Hb and OT-Alb adduct formation may be a valuable biomarker for assessing workplace exposure.