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Equilibrium Dialysis (equilibrium + dialysis)
Selected AbstractsMeasurement of Free Thyroxine Concentration in Horses by Equilibrium DialysisJOURNAL OF VETERINARY INTERNAL MEDICINE, Issue 2 2006Babetta A. Breuhaus The purpose of the study reported here was to validate measurement of free thyroxine (fT4) concentration in equine serum by equilibrium dialysis (fT4D), and to compare values with fT4 concentration measured directly and with total T4 (TT4) concentration. The fT4D, fT74, and TT4 concentrations were measured over a range of values in euthyroid horses and horses made hypothyroid by administration of propylthiouracil (PTU). Concentrations of fT4D (<1.8,83 pmol/L) were consistently higher than those of fT4 (<1,40 pmol/L). There was a significant (P < .001) regression of fT4D on fT4 in 503 samples from normal horses (y = 2.086x - 0.430). In baseline samples from 71 healthy euthyroid horses, fT4 concentration ranged from 6- 21 pmol/L (median, 11 pmol/L; 95% confidence interval [CI]10.5,11.8 pmol/L), and fT4D concentration ranged from 7,47 pmol/L (median, 22 pmol/L; 95% CI 20.9,25.1 pmol/L). Free T4D, fT4, and TT4 concentrations were also measured in 34 ill horses. Horses consuming PTU and ill horses had significantly (P < .05) lower serum concentration of TT4, fT4, and fT4D than did clinically normal, healthy horses. If serum samples from ill horses were further subdivided into samples from horses that lived and samples from horses that died, fT4D concentration was not significantly different in ill horses that lived, compared with that in healthy horses, whereas fT4 concentration was still significantly decreased in ill horses that died (P < 0.001). We conclude that measurement of fT4 concentration by equilibrium dialysis is a valid technique in the horse, and its use may provide improved ability to distinguish nonthyroidal illness syndrome from hypothyroidism in that species. [source] Comparison of Serum-Free Thyroxine Concentrations Determined by Standard Equilibrium Dialysis, Modified Equilibrium Dialysis, and 5 Radioimmunoassays in DogsJOURNAL OF VETERINARY INTERNAL MEDICINE, Issue 3 2004Sara Schachter Measurement of serum-free thyroxine (fT4) concentration provides a more accurate assessment of thyroid gland function than serum thyroxine (T4) or 3,5,3'-triiodothyronine (T3). Techniques for measuring serum fT4 concentration include standard equilibrium dialysis (SED), radioimmunoassay (RIA), and a combination of both (modified equilibrium dialysis [MED]). This study compared results of serum fT4 measurements by means of SED, MED, and 5 RIAs in 30 healthy dogs, 10 dogs with hypothyroidism, and 31 euthyroid dogs with concurrent illness for which hypothyroidism was a diagnostic consideration. Serum fT4 concentrations were comparable when determined by the SED and MED techniques, and mean serum fT4 concentrations were significantly (P<.01) lower in dogs with hypothyroidism than in healthy dogs and euthyroid dogs with concurrent illness. Significant (P < .05) differences in fT4 concentrations were identified among the 5 RIAs and among the RIAs and MED and SED. Serum fT4 concentrations were consistently lower when fT4 was determined by the RIAs, compared with either equilibrium dialysis technique. Serum fT4 concentrations were significantly lower (P < .01) in dogs with hypothyroidism than in healthy dogs for all RIAs; were significantly lower (P < .05) in dogs with hypothyroidism than in euthyroid dogs with concurrent illness for 4 RIAs; and were significantly lower (P < .01) in euthyroid dogs with concurrent illness than in healthy dogs for 4 RIAs. RIAs had the highest number of low serum fT4 concentrations in euthyroid dogs with concurrent illness. This study documented differences in test results among fT4 assays, emphasizing the importance of maintaining consistency in the assay used to measure serum fT4 concentrations in the clinical or research setting. [source] Characterization of site I of human serum albumin using spectroscopic analyses: Locational relations between regions Ib and Ic of site IJOURNAL OF PHARMACEUTICAL SCIENCES, Issue 12 2004Keishi Yamasaki Abstract Site I of human serum albumin is an important and complex region for high-affinity binding of drugs. Equilibrium dialysis showed independent binding of dansyl- L -asparagine (DNSA) and n -alkyl p -aminobenzoates (p -ABEs) to regions Ib and Ic, respectively, in the pH range 6.0,9.0. However, individual binding of DNSA increased with pH in the same range. Binding of the four n -alkyl p -ABEs strongly perturbed the circular dichroism spectrum of bound DNSA, and the effect increased with concentration and the number of carbon atoms in the alkyl moiety. A similar effect was observed by increasing pH from 6.0 to 9.0, a pH range in which human serum albumin is known to undergo the neutral-to-base transition. The spectral changes propose spatial orientation changes of DNSA at region Ib. This proposal was supported by increased fluorescence anisotropy values: n -alkyl p -ABEs binding and the pH-dependent conformational change each restricted the mobility of the naphthalene ring of bound DNSA. Despite the similar effects on the spatial orientation of DNSA, clear differences were observed between the effects of n -alkyl p -ABEs and neutral-to-base transition. The former hardly changed the affinity and maximum fluorescence emission wavelength of bound DNSA; in contrast, the latter significantly affected them. The results give new information about site I and, according to our knowledge, represent a new type of ligand interaction, because the binding site of DNSA could be changed by simultaneous binding of the n -alkyl p -ABEs without affecting the binding constant. © 2004 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 93:3004,3012, 2004 [source] Dimeric 2,2,-Bipyridylruthenium(II) Complexes Containing 2,2,-Bis(1,2,4-triazin-3-yl)-4,4,-bipyridine-Like Bridging Ligands: Syntheses, Characterization and DNA BindingEUROPEAN JOURNAL OF INORGANIC CHEMISTRY, Issue 11 2004Cai-Wu Jiang Abstract Three new bridging ligands 2,2,-bis(1,2,4-triazin-3-yl)-4,4,-bipyridine (btb), 2,2,-bis(1,2,4-triazino[5,6-f]acenaphthylen-3-yl)-4,4,-bipyridine (btapb), 2,2,-bis(5,6-diphenyl-1,2,4-triazin-3-yl)-4,4,-bipyridine (bdptb) and their dimeric 2,2,-bipyridylruthenium(II) complexes [Ru(bpy)2(btb)Ru(bpy)2]4+ (1), [Ru(bpy)2(btapb)Ru(bpy)2]4+ (2), [Ru(bpy)2(bdptb)Ru(bpy)2]4+ (3) have been synthesized and characterized by elemental analysis, fast atom bombardment (FAB) mass spectrometry or electrospray mass spectrometry (ES-MS), 1H NMR and UV/Visible spectroscopy. The binding behavior of these dimeric complexes with calf thymus DNA (CT-DNA) was investigated by electronic absorption spectroscopy, viscosity measurements, and equilibrium dialysis experiments. The hypochromism of the metal-ligand charge transfer (MLCT) band in the electronic absorption spectra of the dinuclear complexes 1, 2, and 3 is 8.7%, 19% and 33%, respectively, with bathochromic shifts of 5, 5 and 14 nm, respectively. The binding constants are 7.5×104M,1, 4.8×105M,1 and 7.6×105M,1, respectively. Increasing the size of the plane of the bridging ligand increases the hydrophobicity of their complexes, leading to stronger binding by the complexes to calf thymus DNA. The effect of increasing concentrations of these novel dimeric ruthenium(II) complexes on the relative viscosities of CT-DNA is less notable than that of well-known intercalators such as [Ru(bpy)2(dppz)]2+. The equilibrium experiments showed that ,,,3 binding is stronger than ,,,3 binding to CT-DNA. This is the first example of a dinuclear complex binding enantioselectively to CT-DNA measured by equilibrium dialysis. The experiments suggest that the three complexes may be DNA groove binders. (© Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2004) [source] Photopolymerizable Hydrogels Made from Polymer-Conjugated Albumin for Affinity-Based Drug Delivery,ADVANCED ENGINEERING MATERIALS, Issue 1-2 2010Liat Oss-Ronen As a drug delivery vehicle, biodegradable albumin hydrogels can combine the high binding capacity of albumin with the structural stability of a polymeric hydrogel network to enable controlled release of small molecules based on both binding affinity and physical interactions. In the present study, we report on the development of a hybrid hydrogel composed of albumin conjugated to poly(ethylene glycol) (PEG) for drug delivery applications where controlled release is accomplished using the natural affinity of the drugs to the serum albumin. Bovine serum albumin was conjugated to PEG-diacrylate having a molecular weight of 1.5, 4, or 10,kDa to form a PEGylated albumin macromolecule (mono-PEGylated or multi-PEGylated). Biodegradable hydrogels were formed from the PEGylated albumin using photopolymerization. Two model drugs, Warfarin and Naproxen, were used for equilibrium dialysis and release experiments from the hydrogels, both having relatively low molecular weights and a known high affinity for albumin. Equilibrium dialysis experiments showed that multi-PEGylation of albumin significantly decreased the drug affinity to the protein compared to non-PEGylated controls, irrespective of the PEG molecular weight. However, the results from drug release experiments showed that mono-PEGylation of albumin did not change its natural affinity to the drug. Comparing the release profiles with a Fickian diffusion model provided strong evidence that hydrogels containing mono-PEGylated albumin exhibited sub-diffusive drug release properties based on the affinity of the drug to the tethered protein. [source] Stereoselective renal tubular secretion of levocetirizine and dextrocetirizine, the two enantiomers of the H1 -antihistamine cetirizineFUNDAMENTAL & CLINICAL PHARMACOLOGY, Issue 1 2008M. Strolin Benedetti Abstract Competition for uptake and/or efflux transporters can be responsible for drug interactions. Cetirizine is mainly eliminated unchanged in urine through both glomerular filtration and tubular secretion. The aim of this study was to investigate whether the eutomer, levocetirizine, and the distomer, dextrocetirizine, have a similar tubular secretion. The renal clearance associated with tubular secretion was calculated from the renal clearance of levocetirizine and dextrocetirizine obtained in a study in healthy volunteers. The values of the unbound fraction in plasma were obtained in an in vitro study of the binding of 14C-cetirizine and 14C-levocetirizine to human plasma proteins using equilibrium dialysis and chiral high-performance liquid chromatography (HPLC) with on-line liquid scintillation counting. The unbound fraction was 0.074 for levocetirizine and 0.141 for dextrocetirizine. The tubular secretion of dextrocetirizine (44.5 mL/min) is higher than that of levocetirizine (23.1 mL/min), which may have consequences for drug interactions at the renal level. The higher tubular secretion for dextrocetirizine may be due to the higher free fraction available for secretion or to a higher affinity for (a) renal transporter(s) mediating the secretion pathway. [source] Lactate concentrations in the rectal lumen in patients in early septic shockACTA ANAESTHESIOLOGICA SCANDINAVICA, Issue 7 2010M. IBSEN Background: Previously, we observed that rectal luminal lactate was higher in non-survivors compared with survivors of severe sepsis or septic shock persisting >24 h. The present study was initiated to further investigate this tentative association between rectal luminal lactate and mortality in a larger population of patients in early septic shock. Methods: A prospective observational multicentre study of 130 patients with septic shock at six general ICU's of university hospitals. Six to 24 h after the onset of septic shock, the concentration of lactate in the rectal lumen was estimated by a 4-h equilibrium dialysis. Dialysate concentrations of lactate were determined using an auto-analyser. Results: The overall 30-day mortality was 32%, with age and Simplified acute physiology scores II and sequential organ failure assessment scores being significantly higher in non-survivors. In contrast, there were no differences in concentrations of lactate in the rectal lumen [2.2 (1.4,4.1) and 2.8 (1.6,5.1) mmol/l (P=0.34)] (medians and 25th,75th percentiles) or arterial blood [2.1 (1.4,4.2) and 2.0 (1.3,3.2) mmol/l (P=0.15)] between non-survivors and survivors. The rectal,arterial difference of the lactate concentration was higher in survivors. There were no differences in blood pressure, noradrenaline dose or central venous oxygen saturation between the groups. Conclusion: In this prospective, observational study of unselected patients with early septic shock, there was no difference in the concentration of lactate in the rectal lumen between non-survivors and survivors. Trial Registration: Clinicaltrials.gov (no: NCT00197938). [source] Physicochemical characterization of carrageenans,A critical reinvestigationJOURNAL OF APPLIED POLYMER SCIENCE, Issue 6 2008Gisela Berth Abstract Kappa-, iota-, and lambda-carrageenan (food grade) were analyzed by static light scattering (MALS in batch mode) in 0.1M NaNO3 at 25 and 60°C, earlier heated up to 90°C or not. At 25°C, there was a strong tendency for a concentration-dependent aggregation in the order lambda < kappa < iota. At 60°C, all samples were molecularly dispersed. The strongly temperature-dependent refractive index increments (equilibrium dialysis) differ. Data interpretation in terms of the wormlike chain model using the Skolnik-Odijk-Fixman approach led to an intrinsic persistence length around 3 to 4 nm and expansion factors as high as 1.5 and above in a thermodynamically good solvent for all three types. Triple-detector HPSEC (DRI, MALS, viscometry) on the three commercial samples plus a degraded (by acidic hydrolysis) kappa-carrageenan in the same solvent/eluant at 60°C yielded a uniform and slightly curved [,]- M relationship for 5 × 103 , M/(g mol) , 3 × 106 and a nearly identical molar mass dependence of the radius of gyration. HPSEC at 25°C on kappa-carrageenan confirmed formation of soluble aggregates. Special emphasis was put on analytical and methodological aspects. The reliability of the experimental data was demonstrated by analogous measurements on dextran calibration standards. © 2008 Wiley Periodicals, Inc. J Appl Polym Sci, 2008 [source] THE FREE RADICAL-SCAVENGING PROPERTY OF CHONDROITIN SULFATE FROM PIG LARYNGEAL CARTILAGE IN VITROJOURNAL OF FOOD BIOCHEMISTRY, Issue 1 2007SHUANG-LI XIONG ABSTRACT This study compared the free radical-scavenging properties of chondroitin sulfate (ChS) from pig laryngeal cartilage and its reduced or sulfonated derivatives. The binding behavior between Cu2+ and ChS and its derivatives, and the interaction between superoxide radical and ChS were studied by fluorescence quenching, equilibrium dialysis, infrared spectra and thermal analysis. Purified ChS inhibited the generation of hydroxyl radical and scavenged superoxide radical in a concentration-dependent manner. Reduced ChS did not scavenge hydroxyl radical and superoxide radical. Sulfonated ChS had no hydroxyl radical scavenging activity but scavenged superoxide radical as strongly as purified ChS. ChS showed strong binding activity with Cu2+ in deionized water but not in 0.01-M HCl. Both reduced ChS and sulfonated ChS did not exhibit such chelating behavior. The structural basis of hydroxyl radical inhibiting of ChS was attributed to a complex of the Cu2+ with the carboxyl group of glucuronic acid residue. The reaction of superoxide radical with the sulfate ester and the carboxyl group may be the basis of superoxide radical scavenging activity of ChS. [source] Liposome transport of hydrophobic drugs: Gel phase lipid bilayer permeability and partitioning of the lactone form of a hydrophobic camptothecin, DB-67JOURNAL OF PHARMACEUTICAL SCIENCES, Issue 1 2008Vijay Joguparthi Abstract The design of liposomal delivery systems for hydrophobic drug molecules having improved encapsulation efficiency and enhanced drug retention would be highly desirable. Unfortunately, the poor aqueous solubility and high membrane binding affinity of hydrophobic drugs necessitates extensive validation of experimental methods to determine both liposome loading and permeability and thus the development of a quantitative understanding of the factors governing the encapsulation and retention/release of such compounds has been slow. This report describes an efflux transport method using dynamic dialysis to study the liposomal membrane permeability of hydrophobic compounds. A mathematical model has been developed to calculate liposomal membrane permeability coefficients of hydrophobic compounds from dynamic dialysis experiments and partitioning experiments using equilibrium dialysis. Also reported is a simple method to study the release kinetics of liposome encapsulated camptothecin lactone in plasma by comparing the hydrolysis kinetics of liposome entrapped versus free drug. DB-67, a novel hydrophobic camptothecin analogue has been used as a model permeant to validate these methods. Theoretical estimates of DB-67 permeability obtained from the bulk solubility diffusion model and the "barrier-domain" solubility diffusion model are compared to the experimentally observed value. The use of dynamic dialysis in drug release studies of liposome and other nanoparticle formulations is further discussed and experimental artifacts that can arise without adequate validation are illustrated through simulations. © 2007 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 97:400,420, 2008 [source] High-throughput determination of the free fraction of drugs strongly bound to plasma proteinsJOURNAL OF PHARMACEUTICAL SCIENCES, Issue 4 2004Joachim Schuhmacher Abstract Quantification of protein binding of new chemical entities is an important early screening step during drug discovery and is of fundamental interest for the estimation of safety margins during drug development. In this publication, we describe the development of a new high-throughput assay for the determination of the free drug fraction in plasma (fu). The new technique is an enhancement of the previously published erythrocytes partition method. It is based on the distribution of drugs between plasma water, plasma proteins, and solid-supported lipid membranes (Transil®). The execution of protein binding studies by partitioning is dramatically simplified by substituting erythrocytes with commercially available Transil® beads, and makes the method particularly suitable for high-throughput studies. Eight Bayer compounds from different compound classes covering a wide range of lipophilicities (log P,=,1.9,5.6) and fu values (0.018,35%) were selected for validation of the assay. The results obtained by the new method and by either the erythrocytes partitioning technique or more conventional methods (ultrafiltration and equilibrium dialysis) are identical, confirming that the new method produces valid results even for drugs that are strongly bound to plasma proteins. Precision and accuracy of the data in the cases of very low and high fu values are comparable, indicating that the method is especially suited for highly lipophilic drugs that tend to adsorb to surfaces compared with other methods, like ultrafiltration or equilibrium dialysis, that may produce biased data. The method is also useful for the determination of binding parameters and the pH dependence of fu. In summary, this assay is well suited for high-throughput determination of protein binding during drug discovery and for extended protein binding studies during drug development. © 2004 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 93: 816,830, 2004 [source] Characterization of drug,protein interactions in blood using high-performance affinity chromatographyJOURNAL OF SEPARATION SCIENCE, JSS, Issue 5-6 2009David S. Hage Abstract The binding of drugs with proteins in blood, serum, or plasma is an important process in determining the activity, distribution, rate of excretion, and toxicity of drugs in the body. High-performance affinity chromatography (HPAC) has received a great deal of interest as a means for studying these interactions. This review examines the various techniques that have been used in HPAC to examine drug,protein binding and discusses the types of information that can be obtained through this approach. A comparison of these techniques with traditional methods for binding studies (e.g., equilibrium dialysis and ultrafiltration) will also be presented. The use of HPAC with specific serum proteins and binding agents will then be discussed, including HSA and ,1 -acid glycoprotein (AGP). Several examples from the literature are provided to illustrate the applications of such research. Recent developments in this field are also described, such as the use of improved immobilization techniques, new data analysis methods, techniques for working directly with complex biological samples, and work with immobilized lipoproteins. The relative advantages and limitations of the methods that are described will be considered and the possible use of these techniques in the high-throughput screening or characterization of drug,protein binding will be discussed. [source] In vitro bioavailability of calcium and iron from selected green leafy vegetables,JOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 13 2006Sheetal Gupta Abstract The objective of the present investigation was to analyze the relative influence of oxalic acid, phytic acid, tannin and dietary fiber on in vitro availability of iron and calcium from green leafy vegetables (GLV). Thirteen GLV were selected and analyzed for iron, calcium, oxalic acid, phytic acid, tannin and dietary fiber contents using standard methods. The bioavailability of calcium and iron in the GLV was estimated by equilibrium dialysis. Oxalic acid content was less than 1 g kg,1 in four greens and ranged between 1.22 to 11.98 g kg,1 in the remaining. Dietary fiber ranged from 19.5 to 113.7 g kg,1. Tannin content ranged between 0.6138 and 2.1159 g kg,1 with the exception of two GLV that had 0.1332 and 14.8619 g kg,1. Four GLV were found to have approximately 40% bioavailable iron, while the others were in the range of 6,30%. In vitro available calcium was less than or equal to 25% in eight GLV and between 34% and 52% in five GLV. Multiple regression analysis revealed that these factors together accounted for 53% (r2 = 0.53) and 45% (r2 = 0.45) inhibition of iron and calcium absorption, respectively. These findings infer that calcium and iron availability is influenced by the constituents present in the GLV. Copyright © 2006 Society of Chemical Industry [source] Influence of Isoflurane General Anesthesia or Anesthesia and Surgery on Thyroid Function Tests in DogsJOURNAL OF VETERINARY INTERNAL MEDICINE, Issue 1 2009M.A. Wood Background: Anesthesia and surgery affect thyroid function tests in humans but have not been studied in dogs. Hypothesis: Anesthesia and anesthesia with surgery will affect thyroid function tests in dogs. Animals: Fifteen euthyroid dogs. Methods: Prospective, controlled, interventional study. Dogs were assigned to one of 3 groups: control, general anesthesia, and general anesthesia plus abdominal exploratory surgery. Dogs in the anesthesia and surgery groups were premedicated with acepromazine and morphine, induced with propofol, and maintained on isoflurane. Samples for measurement of serum thyroxine (T4), free T4 (fT4) by equilibrium dialysis, triiodothyronine (T3), reverse T3 (rT3), and thyroid-stimulating hormone concentrations were collected from each dog immediately before premedication, at multiple times during anesthesia, surgery, 4, 8, 12, 24, 36, and 48 hours after anesthesia, once daily for an additional 5 days, and once 14 days after anesthesia. Sampling was performed at identical times in the control group. Results: Serum T4 decreased significantly from baseline in the surgery and anesthesia groups compared with the control group at 0.33 (P= 0.043) and 1 hour (P= 0.018), and 2 (P= 0.031) and 4 hours (P= 0.037), respectively, then increased significantly in the surgery group compared with the control group at 24 hours (P= 0.005). Serum T3 decreased significantly from baseline in the anesthesia group compared with the control group at 1 hour (P= 0.034). Serum rT3 increased significantly from baseline in the surgery group compared with the control and anesthesia groups at 8 (P= 0.026) and 24 hours (P= 0.0001) and anesthesia group at 8, 12, 24, and 36 hours (P= 0.004, P= 0.016, P= 0.004, and P= 0.014, respectively). Serum fT4 increased significantly from baseline in the surgery group compared to the control at 24 hours (P= 0.006) and at day 7 (P= 0.037) and anesthesia group at 48 hours (P= 0.023). Conclusions and Clinical Importance: Surgery and anesthesia have a significant effect on thyroid function tests in dogs. [source] Measurement of Free Thyroxine Concentration in Horses by Equilibrium DialysisJOURNAL OF VETERINARY INTERNAL MEDICINE, Issue 2 2006Babetta A. Breuhaus The purpose of the study reported here was to validate measurement of free thyroxine (fT4) concentration in equine serum by equilibrium dialysis (fT4D), and to compare values with fT4 concentration measured directly and with total T4 (TT4) concentration. The fT4D, fT74, and TT4 concentrations were measured over a range of values in euthyroid horses and horses made hypothyroid by administration of propylthiouracil (PTU). Concentrations of fT4D (<1.8,83 pmol/L) were consistently higher than those of fT4 (<1,40 pmol/L). There was a significant (P < .001) regression of fT4D on fT4 in 503 samples from normal horses (y = 2.086x - 0.430). In baseline samples from 71 healthy euthyroid horses, fT4 concentration ranged from 6- 21 pmol/L (median, 11 pmol/L; 95% confidence interval [CI]10.5,11.8 pmol/L), and fT4D concentration ranged from 7,47 pmol/L (median, 22 pmol/L; 95% CI 20.9,25.1 pmol/L). Free T4D, fT4, and TT4 concentrations were also measured in 34 ill horses. Horses consuming PTU and ill horses had significantly (P < .05) lower serum concentration of TT4, fT4, and fT4D than did clinically normal, healthy horses. If serum samples from ill horses were further subdivided into samples from horses that lived and samples from horses that died, fT4D concentration was not significantly different in ill horses that lived, compared with that in healthy horses, whereas fT4 concentration was still significantly decreased in ill horses that died (P < 0.001). We conclude that measurement of fT4 concentration by equilibrium dialysis is a valid technique in the horse, and its use may provide improved ability to distinguish nonthyroidal illness syndrome from hypothyroidism in that species. [source] Comparison of Serum-Free Thyroxine Concentrations Determined by Standard Equilibrium Dialysis, Modified Equilibrium Dialysis, and 5 Radioimmunoassays in DogsJOURNAL OF VETERINARY INTERNAL MEDICINE, Issue 3 2004Sara Schachter Measurement of serum-free thyroxine (fT4) concentration provides a more accurate assessment of thyroid gland function than serum thyroxine (T4) or 3,5,3'-triiodothyronine (T3). Techniques for measuring serum fT4 concentration include standard equilibrium dialysis (SED), radioimmunoassay (RIA), and a combination of both (modified equilibrium dialysis [MED]). This study compared results of serum fT4 measurements by means of SED, MED, and 5 RIAs in 30 healthy dogs, 10 dogs with hypothyroidism, and 31 euthyroid dogs with concurrent illness for which hypothyroidism was a diagnostic consideration. Serum fT4 concentrations were comparable when determined by the SED and MED techniques, and mean serum fT4 concentrations were significantly (P<.01) lower in dogs with hypothyroidism than in healthy dogs and euthyroid dogs with concurrent illness. Significant (P < .05) differences in fT4 concentrations were identified among the 5 RIAs and among the RIAs and MED and SED. Serum fT4 concentrations were consistently lower when fT4 was determined by the RIAs, compared with either equilibrium dialysis technique. Serum fT4 concentrations were significantly lower (P < .01) in dogs with hypothyroidism than in healthy dogs for all RIAs; were significantly lower (P < .05) in dogs with hypothyroidism than in euthyroid dogs with concurrent illness for 4 RIAs; and were significantly lower (P < .01) in euthyroid dogs with concurrent illness than in healthy dogs for 4 RIAs. RIAs had the highest number of low serum fT4 concentrations in euthyroid dogs with concurrent illness. This study documented differences in test results among fT4 assays, emphasizing the importance of maintaining consistency in the assay used to measure serum fT4 concentrations in the clinical or research setting. [source] Liposome/water lipophilicity: Methods, information content, and pharmaceutical applicationsMEDICINAL RESEARCH REVIEWS, Issue 3 2004Georgette Plemper van Balen Abstract This review discusses liposome/water lipophilicity in terms of the structure of liposomes, experimental methods, and information content. In a first part, the structural properties of the hydrophobic core and polar surface of liposomes are examined in the light of potential interactions with solute molecules. Particular emphasis is placed on the physicochemical properties of polar headgroups of lipids in liposomes. A second part is dedicated to three useful methods to study liposome/water partitioning, namely potentiometry, equilibrium dialysis, and 1H-NMR relaxation rates. In each case, the principle and limitations of the method are discussed. The next part presents the structural information encoded in liposome/water lipophilicity, in other words the solutes' structural and physicochemical properties that determine their behavior and hence their partitioning in such systems. This presentation is based on a comparison between isotropic (i.e., solvent/water) and anisotropic (e.g., liposome/water) systems. An important factor to be considered is whether the anisotropic lipid phase is ionized or not. Three examples taken from the authors' laboratories are discussed to illustrate the factors or combinations thereof that govern liposome/water lipophilicity, namely (a) hydrophobic interactions alone, (b) hydrophobic and polar interactions, and (c) conformational effects plus hydrophobic and ionic interactions. The next part presents two studies taken from the field of QSAR to exemplify the use of liposome/water lipophilicity in structure,disposition and structure,activity relationships. In the conclusion, we summarize the interests and limitations of this technology and point to promising developments. © 2004 Wiley Periodicals, Inc. Med Res Rev, 24, No. 3, 299,324, 2004 [source] Study of factors affecting binding of zinc with albumin at physiological zinc concentrationsBIOFACTORS, Issue 3 2004V.V. Agte Abstract Albumin has very high affinity for many organic and inorganic compounds that may influence albumin bound Zn (ABZn). To get insight of these molecular interactions, the effect of riboflavin, nicotinic acid, thiamine, folic acid, pyruvic acid and glucose on ABZn were studied. The ABZn was separated from the unbound zinc using equilibrium dialysis and estimated using atomic absorption spectrometer. At therapeutic zinc concentrations, folic acid and thiamine significantly enhanced the ABZn (p < 0.010), while nicotinic acid inhibited zinc binding to albumin. Folic acid was found to enhance the ABZn also at lower zinc concentrations representing physiological levels of plasma zinc (138,150 micromoles) (p < 0.05). [source] New method for the simultaneous estimation of intrinsic hepatic clearance and protein binding by matrix inhibitionBIOPHARMACEUTICS AND DRUG DISPOSITION, Issue 1 2008Takahide Uchimura Abstract The purpose of this study was to develop a method for estimating the hepatic clearance (CLh) without using a protein binding test. This method allows the simultaneous evaluation of the intrinsic hepatic clearance (CLint) with a correction for microsomal binding, and the free fraction in the serum (fu). It uses the decrease in metabolic velocity achieved by decreasing the free fraction of a compound in the incubation mixture (fuinc) by the addition of serum, and by changing the microsomal protein concentration. This method is denoted as the ,matrix inhibition method', because it uses the inhibition of the metabolic velocity by the incubation matrix. The metabolic rates of eight compounds (diazepam, imipramine, warfarin, and compounds A,E) were evaluated under several incubation conditions using rat serum and microsomes. The correlation of CLint evaluated using the method and using equilibrium dialysis after the CLint was corrected for microsomal binding was r,=,0.968. The correlation of fu,·,CLint was r,=,0.996. Although the method required a high enough fu and fumicrosomes difference among the reaction conditions for each compound, it could evaluate CLint and fu simultaneously and easily by adding additional reaction conditions to the metabolic stability tests performed in ADME screening. Copyright © 2007 John Wiley & Sons, Ltd. [source] | ||||||||||||||||