SEARCH · PubMed Health
Explore indexed PubMed citations for clinical trials, systematic reviews and public health research. Read source abstracts and follow each citation to its original PubMed record.
Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.
A simple device for the production of serum ultrafiltrates under controlled conditions of pH is described. The device has been tested for the values obtained in normal human subjects are reported.
Two beta-thymosins are expressed in most mammalian tissues. We detected small amounts of a third peptide in extracts of rabbit spleen. The portion of this peptide increased when the tissue was first frozen and then thawed at 4 degrees C. Small amounts of the peptide are also present in cells from suspension cultures homogenized immediately in diluted perchloric acid. By means of amino acid analysis and MALDI-mass spectroscopy this peptide was identified to be a C-terminally truncated form of thymosin beta10. Having studied the formation in more detail we found that after a 4-h thaw at 4 degrees C all thymosin beta10 was truncated to thymosin beta10(1-41), which was further degraded during the next 20 h. On the other hand, thymosin beta4Ala, the second beta-thymosin being present in rabbit spleen, was not truncated or degraded even after 22 h. It might be possible that in vivo a truncated form of thymosin beta10 is formed by a carboxydipeptidase while thymosin beta4Ala is rather stable against proteolytic modification. By using a newly designed ultrafiltration assay, we determined the dissociation constants of the complexes of G-actin and these three beta-thymosins to be 0.28, 0.72, and 0.94 microM for thymosin beta4Ala, beta10, and thymosin beta10(1-41), respectively. The complex with beta4Ala is unambiguously more stable than the complex with beta10 or beta4 (0.81 microM). The change in the dissociation constant generated by the truncation of the two C-terminal amino acid residues of beta10 is small but statistically significant. This demonstrates that even the very last amino acid residues at the C-terminus of beta-thymosins are involved in the interaction with G-actin.
The purification of the human peptide hormone guanylin 22-115 from blood ultrafiltrate (hemofiltrate, HF) was achieved using matrix-assisted laser desorption/ionisation mass spectrometry (MALDI-MS) as the assay system. Screening a peptide bank generated from 5000 1 HF guanylin 22-115 was detected by its molecular mass when adequate conditions for MALDI-MS analysis were chosen. The sensitivity was even better than of the established biological assay system. In addition, the susceptibility towards solvents and salts is strongly reduced. 1.2 mg of the peptide hormone was purified from 10% of the starting material.
The effects that wastewater quality and mode of operation have on the performance of an asymmetric, hollow fiber, polysulfone, ultrafiltration (UF) membrane with a molecular weight cutoff of 100,000 Daltons were investigated. Performance was assessed through monitoring membrane flux, transmembrane pressure, effluent biochemical oxygen demand, and operational cost of the experimental system while treating filtered secondary, secondary, and filtered primary effluents. Fluxes achieved for filtered secondary (129-173 l/m2 h), secondary (101-158 l/m2 h), and filtered primary (20-41 l/m2 h) effluents were compared to those obtained at three other locations where similar UF systems were operated. A conceptual model of the impact of an insufficient backwash and of operating the UF system at constant flux on membrane performance is presented to explain the differences in fluxes. Employing pre-membrane granular filtration to remove a portion of the problematic particles in secondary effluent prior to UF led to optimal operational conditions. The costs associated with the operation of pre-membrane granular filtration were offset by the increase in production achieved. Although the use of recirculation could increase maintainable flux when treating a concentrated feed (e.g., filtered primary effluent), the associated costs were high. Improved UF performance was found to result from allowing flux to decline naturally, rather than using a constant flux mode of operation. The effluents produced when filtered secondary and secondary effluents were the feeds would be equivalent to an oxidized, coagulated, clarified, and filtered wastewater as per Title 22 California Wastewater Reclamation Criteria.
One of the most common problems encountered in water treatment applications of membranes is fouling. Natural organic matter (NOM) represents a particularly problematic foulant. Membranes may be fouled by relatively hydrophilic and/or hydrophobic NOM components, depending on NOM characteristics, membrane properties, and operating conditions. To maximize flux recovery for an NOM-fouled ultrafiltration membrane (NTR 7410), chemical cleaning and hydraulic rinsing with a relatively high cross-flow velocity were investigated as cleaning strategies. The modification of the membrane surface with either an anionic or a cationic surfactant was also evaluated to minimize membrane fouling and to enhance NOM rejection. Foulants from a hydrophobic NOM source (Orange County ground water (OC-GW)) were cleaned more effectively in terms of permeate flux by acid and caustic cleanings than foulants from a relatively hydrophilic NOM source (Horsetooth surface water (HT-SW)). An anionic surfactant (sodium dodecyl sulfate (SDS)) was not effective as a cleaning agent for foulants from either hydrophobic or hydrophilic NOM sources. High ionic strength cleaning with 0.1 M NaCl was comparatively effective in providing flux recovery for NOM-fouled membranes compared to other chemical cleaning agents. Increased cross-flow velocity and longer cleaning time influenced the efficiency of caustic cleaning, but not high ionic strength cleaning. The membrane was successfully modified only with the cationic surfactant; however, enhanced NOM rejection was accompanied by a significant flux reduction.
Ultrafiltration (UF) membranes from polysulfone (PSf) were functionalized by heterogeneous photo-initiated graft copolymerization of acrylic acid (AA). With radiation susceptible PSf, only proper selection of the UV energy (lambda > 350 nm; for selective excitation of the photoinitiator) yielded membranes with preserved UF barrier layer. Possibilities for adjusting structure and morphology of the graft polymer (g-PAA) layer by variation of functionalization parameters such as AA concentration and UV irradiation time were investigated. Very long grafted chains (Mw > 10(5) g mol(-1)) at varied grafting density (GD = 0.01 ... 1.2 nmol cm(-2), relative to the outer surface area) were obtained. Partial penetration of the UF barrier layer by g-PAA was verified. Covalent immobilization of bovine serum albumin (BSA), gamma-globulin (gamma-Gl) and alkaline phosphatase (APh) was achieved by coupling with a water soluble carbodiimide. Bound BSA and gamma-Gl amounts were up to gamma = 10 microg cm(-2), for membranes accessible only from the outer surface thus not using the entire pore volume. Locally addressed covalent protein immobilization after photo-patterning the PSf surface could be visualized with a fluorescent FITC-BSA conjugate. A strong salt effect onto immobilized APh activity (increase with NaCl concentration) was observed, indicating internal transport/accessibility limitations in the g-PAA layer. Correlations between PAA structure (Mw, GD) and accessibility (from BSA or gamma-G1 binding and APh activity) could be established. The 'tentacle' g-PAA functionalized PSf UF membranes having preserved UF barrier and, e.g., with surface-bound receptors will find application in cell cultures under diffusion or perfusion conditions.
The study aimed to develop simple assays to study aluminium-ligand interactions in natural/biological systems where equilibrium is rarely reached and thus where the initial seconds or hours of interactions are important. The immediate and non-equilibrium precipitation of aluminium hydroxide, in aqueous solution at neutral pH, was therefore studied by laser light scattering (diffraction), ultrafiltration and centrifugation. The interaction of weak ligands, present in the gastrointestinal lumen, on the precipitation of aluminium hydroxide was also investigated. The initial kinetics and particle sizes of precipitated aluminium hydroxide were sensitive to a number of external factors, including the presence of weak ligand (bicarbonate), sheer force (stirring), electrolyte concentration and initial (i.e. added) aluminium concentration. However, after a few seconds (no weak ligand), or several hundred seconds (with weak ligand), the subsequent observed changes to the solid phase were of small magnitude and occurred slowly. Thus, a 25-min window, within 5 and 30 min of pH adjustment, can be used to study the interactions of aluminium-ligand. This may approximate better to most natural systems where unperturbed aluminium-ligand equilibrium must rarely exist.
Imbalance between cardiac oxygen supply and demand may trigger cardiac events in already vulnerable hemodialysis (HD) patients. We studied the effect of ultrafiltration (UF) and HD in nine chronic HD patients by continuously measuring blood volume (BV; by Critline), blood pressure (BP; by Portapres), and changes in hemodynamics (Modelflow) during isolated UF (iUF) of 500 mL in 30 minutes and subsequent HD combined with UF (HD + UF). Aortic pressure was reconstructed from finger pressure. Changes in cardiac oxygen supply were assessed by calculating the area under the aortic pressure curve during diastole (diastolic pressure time index [DPTI]). Changes in cardiac oxygen demand were assessed by calculating systolic pressure time index (SPTI). BV decreased 4.0% +/- 1.8% during UF and 7.3% +/- 3.3% during HD + UF (both P < 0.01). Systolic BP did not change; diastolic and mean BP increased 11 +/- 7.4 and 11 +/- 8.4 mm Hg during iUF, respectively (both P < 0.01), and stabilized during HD + UF. Overall pulse pressure decreased 19 +/- 11.1 mm Hg (P < 0.01). Heart rate increased 13 +/- 11 beats/min (P < 0.01) and systemic vascular resistance increased 59% +/- 51% (P < 0. 01), whereas stroke volume and cardiac output (CO) decreased by 40% +/- 17% and 30% +/- 13%, respectively (both P < 0.01). Both cardiac oxygen supply (DPTI) and demand (SPTI) increased during iUF, and both decreased during HD + UF. By the end of the procedure, DPTI/SPTI ratio had increased 9% +/- 8% (P < 0.05). Changes in CO correlated closely to changes in BV. Despite large changes in hemodynamics during uncomplicated UF and HD, the balance between cardiac oxygen supply and demand (DPTI/SPTI ratio) did not decrease, but improved slightly.
A patient is presented who after 2 years of hemodialysis showed all of the features of congestive cardiomyopathy to a very severe degree: dilation of all cardiac compartments, increased left ventricular mass, low ejection fraction, diastolic disturbances, third- to fourth-degree mitral and tricuspid regurgitation, ascites, and low blood pressure. All of these abnormalities gradually but completely disappeared during 5 months of persistent ultrafiltration during or between dialysis sessions. It was concluded that chronic fluid overload was a major factor in the cardiac disease of this patient. Unrecognized hidden fluid overload has long been known (but also neglected), and its prevention deserves top priority in chronic dialysis patients.
The novel anti-tumour agent 5,6-dimethylxanthenone-4-acetic acid (DMXAA) is a highly protein bound drug with narrow therapeutic window. We report a simple HPLC method with fluorimetric detection for the determination of free DMXAA concentration in human plasma. Sample preparation involves the ultrafiltration of plasma by a Centrisart device for 30 min at 2000 g and extraction with acetonitrile: methanol mixture. The method was validated with respect to recovery, selectivity, linearity, precision, and accuracy. Calibration curves for DMXAA were constructed at the concentration range of 0.5-40 microM in blank plasma and phosphate buffer. The difference between the theoretical and calculated concentration and the relative standard deviation were less than 10% at all quality control (QC) concentrations. The HPLC method has been used for the analysis of preclinical studies.
Sameridine is a new candidate drug with both local anaesthetic and analgesic properties. The free concentration of sameridine in blood plasma was determined by coupled-column liquid chromatography. Following adjustment of the pH and the temperature of the plasma samples, the free fraction was prepared by ultrafiltration. The coupled-column liquid chromatographic system consisted of a reversed-phase column, a cation-exchange extraction column and a cation-exchange analytical column. Sameridine was detected by UV determination at 205 nm and the system showed high selectivity. The limit of quantification was 1 nM and the within-day precision was 4.6% (R.S.D., n = 10).
Chiral HPLC methods were developed and utilized for the simultaneous determination of plasma protein binding of enantiomers of two racemic aminohydantoin compounds. Reversed-phase HPLC with the use of a polysaccharide-type chiral stationary phase column was employed for the separation and quantitation of the enantiomers of the two compounds with detection limits in the range 5-10 ng/ml in the plasma matrix. The chiral HPLC methods were selective, sensitive and reproducible. The R and S enantiomers of both compounds were baseline-resolved under the chromatographic conditions employed. Ultrafiltration techniques were applied to determining the plasma protein binding for each enantiomer in rat, dog and human plasma. The results clearly show stereoselective binding of the two enantiomers of each compound with higher protein binding of the R enantiomer than the S enantiomer in rat, dog and human plasma. Binding association constants were also determined to be in the range 1.01-14.0 x 10(4) M(-1) at 37 degrees C. Both the protein binding percentage and binding association constant were enantioselective and species-dependent. Such information is important for a clear understanding of the differences in biological activity as well as in pharmacokinetic and pharmacodynamic properties between the two enantiomers of each compound in the drug discovery and development process.
The aim of this study was to describe a high-performance liquid chromatography (HPLC) assay for the determination of cefepime and cefpirome in human serum without changing chromatographic conditions. The assay consisted to measure cefepime and cefpirome which were unbound to proteins having a molecular mass of 10,000 or more by ultrafiltration followed by HPLC with a Supelcosil ABZ+ column and UV detection at a wavelength of 263 nm. The assay was been found to be linear and has been validated over the concentration range 200 to 0.50 microg/ml for both cefepime and cefpirome, from 200 microl serum, extracted. In future, the assay will support therapeutic drug monitoring for cefepime and cefpirome in neutropenic patients in correlation with microbiological parameters such as MIC90 (minimal inhibitory concentration of antibiotic which kills 90% of the initial bacterial inoculum) and clinical efficacy.
A sensitive and specific procedure is described for the determination of therapeutically relevant concentrations of L-dopa in plasma and plasma ultrafiltrates (free fraction) by high performance liquid chromatography with electrochemical detection. In plasma samples from healthy adult subjects (n = 15) spiked with L-dopa (500 micrograms l-1) the free fraction averaged 76 +/- = 8% (range 61-84%). Free fraction values increased by 38% with increasing plasma concentrations of L-dopa from 100-5000 micrograms 1-1. L-dopa free fraction was not affect by the presence of 3-O-methyl-dopa at concentrations up to 10,000 micrograms l-1.
The binding of iralukast to plasma (or serum) proteins and to erythrocytes was studied in vitro, at +37 degrees C, using the erythrocyte partitioning method (EPM) and/or ultrafiltration (UF) with 14C-labelled iralukast. Iralukast was highly bound in human and animal serum (>99%). Similar bound fraction values were obtained with the two methods: in whole human plasma (or serum) 99.8% (EPM) and 99.9% (UF), in albumin solution 99.8% (EPM and UF), in high density lipoprotein solution 97.3% (EPM) and 98.3% (UF), and in low density lipoprotein solution 97.2% (EPM) and 98.8% (UF). Moreover, the erythrocyte partitioning method allowed the evaluation of other binding parameters. The binding capacity (l/micromol) of proteins equalled 35 for low density lipoproteins, 3.6 for high density lipoproteins, 1.0 for albumin, 0.78 for alpha-1-acid glycoprotein, and 0.03 for gamma globulins. In whole blood, iralukast was distributed between plasma and erythrocytes in the proportion (%) 90/10. At physiological protein concentrations, iralukast was primarily bound to albumin (79%).
Microdialysis and ultrafiltration are complementary sampling techniques that facilitate acquisition of data in awake, freely moving animals. Because the necessity of blood removal is eliminated, sampling frequency is not limited by animal size. The samples obtained by these techniques usually require no processing for analysis.
A rapid HPLC method was developed for quantification of unbound evernimicin in human plasma. Protein-free samples prepared by ultrafiltration were injected directly onto a polymeric reversed-phase column and the eluent monitored at 302 nm. Evernimicin that eluted within 3.5 min was well resolved from endogenous components. Linearity was established between peak height and evernimicin concentration from 25 to 2500 ng/ml. Assay precision (C.V.) was within 5% while bias was no greater than 3%. This method has been used for the ex vivo assessment of evernimicin protein binding in human plasma from safety and tolerance as well as liver dysfunction and renal insufficiency studies.
Goat whey was hydrolysed by pepsin in an ultrafiltration membrane enzymic reactor coupled with a 30 kDa mineral membrane. Peptides collected in the permeate were resolved using reversed-phase HPLC. Their sequences were determined by amino acid analysis, second order derivative spectra analysis and mass spectrometry. Owing to the resistance of beta-lactoglobulin (beta-lg) towards pepsin, the majority of peptides identified were derived from alpha-lactalbumin (alpha-la). Pepsin showed a broad specificity of hydrolysis sites and generated a wide range of products from dipeptides to very large peptides containing disulphide bridges. The molecular masses of peptides resulting from alpha-la degradation were between 150 and 6900 Da: 36% were < 600 Da. 24% were 600-2000 Da and 40% were > 2000 Da.