Elimination Of Small Molecules Research Articles

OPTIMIZED CONVECTIVE VOLUME IN ONLINE HEMODIAFILTRATION

Hemodiafiltration (HDF) adds convective elimination of middle molecules (MM), proportional to filtered volume (Vconv) on the top of diffusion-based elimination of small molecules (SM) by conventional hemodialysis (HD). Studies, both observational and randomized controlled ones, performed so far generally indicated positive impact of higher Vconv on all-cause mortality in HDF patients, although the magnitude of Vconv at which HDF becomes apparently superior to HD differed widely among the studies. Also the issue of a suitable anthropometric parameter by which the Vconv should be normalized has not yet been solved. Data from the ESHOL study seems to indicate that patient´s body surface area (BSA) could well be used for this—mortality was decreasing with increasing Vconv/BSA with a bottom plateau starting at about 15 L/m2. We have therefore devised a formula and a graph for determination of Vconv which fulfils the requirement Vconv/BSA= 15. Assuming maximum feasible and safe filtration fraction QF/QB= 0.3, the Vconv actually defines the necessary blood flow (QB) to reach Vconv/BSA= 15 as QB=15·BSA/(0.3·t) (t – session time). It is also possible to check what QB is needed in terms of sufficient diffusion-based transport (target Kt/V) and compare both QB values to see if adequate combined elimination of both SM and MM can be achieved at the same time and under what conditions, respectively.

Different elimination patterns of β‐trace protein, β2‐microglobulin and cystatin C in haemodialysis, haemodiafiltration and haemofiltration

Objective. Low molecular mass proteins (LMMP) are putative uraemic toxins, but their elimination is negligible in standard haemodialysis (HD). In this study, we used β2‐microglobulin, cystatin C and β‐trace protein, which differ in molecular mass and charge, to characterize the elimination patterns of three different dialysis modalities. Material and methods. Plasma samples were obtained at the start, 30 min after the start, at the end of the dialysis treatment and 30 min after termination of the dialysis session. Seventeen patients were treated with low‐flux HD, 13 with post‐dilution haemodiafiltration (HDF) and 8 with pre‐dilution haemofiltration (HF). The changes in concentrations of the three LMMPs were monitored and expressed as percentages of the concentrations at the start of treatments. Results. Conventional HD with low‐flux membranes showed a high elimination of small molecules (urea and creatinine), but did not reduce the levels of the three LMMPs studied. During HDF and HF, there was a significant decrease in the plasma levels of cystatin C (to 28 % and 44 %, respectively) (p<0.001) and of β2‐microglobulin (to 23 % and 33 %, respectively) (p<0.001). However, the level of β‐trace protein was significantly reduced (to 65 %) only after HDF. Conclusions. The three dialysis modalities showed significantly different elimination patterns for the LMMPs studied. Elimination of β‐trace protein was lower than those of cystatin C and β2‐microglobulin both in HDF and HF. β‐trace protein was only moderately eliminated by HDF and not at all by HF, and may be a useful marker in the evaluation of different convective therapies.

Two-dimensional Blue Native/SDS Gel Electrophoresis of Multiprotein Complexes from Helicobacter pylori

The study of protein interactions constitutes an important domain to understand the physiology and pathogenesis of microorganisms. The two-dimensional blue native/SDS-PAGE was initially reported to analyze membrane protein complexes. In this study, both cytoplasmic and membrane complexes of a bacterium, the strain J99 of the gastric pathogen Helicobacter pylori, were analyzed by this method. It was possible to identify 34 different proteins grouped in 13 multiprotein complexes, 11 from the cytoplasm and two from the membrane, either previously reported partially or totally in the literature. Besides complexes involved in H. pylori physiology, this method allowed the description of interactions involving known pathogenic factors such as (i) urease with the heat shock protein GroEL or with the putative ketol-acid reductoisomerase IlvC and (ii) the cag pathogenicity island CagA protein with the DNA gyrase GyrA as well as insight on the partners of TsaA, a peroxide reductase/stress-dependent molecular chaperone. The two-dimensional blue native/SDS-PAGE combined with mass spectrometry is a potential tool to study the differences in complexes isolated in various situations and also to study the interactions between bacterial and eucaryotic cell proteins.

Polymer degradation by elimination of small molecules

AbstractAn outline of a method of deriving the kinetic equations for polymer degradation by the elimination of low molecular weight compounds is presented. The method is based on the stochastic approach. The main ideas are that the degradation occurs via the zip mechanism and that the initiating step, i.e. the formation of an olefinic site in the polymer chain, takes place at random. Three mechanisms have been investigated in this study: (i) ordinary zip mechanism; (ii) autocatalytic mechanism with immediate zip growth and catalyzed initiation; (iii) autocatalytic mechanism with gradual zip growth. The kinetic equations derived have been used in kinetic studies of the dehydrochlorination of PVC in inert, HCl, and oxidative atmospheres, and of the degradation of polyvinyl esters, polyvinyl bromide and polyvinylidene chloride. Kinetic parameters for the degradation of individual polymers have been obtained.

A Review of Thermally Controlled Covalent Bond Formation in Polymer Chemistry

Abstract The purpose of this paper is to review known art in the area of thermally controlled covalent linkage formation for use in thermally reversible polymer systems. Only 1 :1 adduct forming reactions are considered in this paper; e.g., condensation reactions are not discussed because these reactions are controlled via the addition or elimination of small molecules. In the case of an exothermic process, covalent bonds would form at lower temperatures and disassemble at higher temperatures. Linkage formation could ideally be extended to either linear polymerizations or crosslinking schemes, as outlined in Fig. 1.

Stereochemistry of organic ions in the gas phase: A review

The stereochemistry of organic ions in the gas phase can be regarded from two different points of view: (i) stereoselectivity in ion formation and (ii) stereospecifity of ion fragmentations. Fast ionization by electron or photon impact shows little stereoselection. Differences in the ionization energies and cross sections between stereoisomers are generally small, save for a few exceptions. Proton or larger ion transfer, as employed in chemical ionization mass spectrometry, gives more possibilities for stereoselection. Bi- or polyfunctional molecules can capture the proton in a hydrogen-bond stabilized [M + H]+ ion, which is feasible only with a favourable spatial orientation of the chelating groups. Adduct ions [M + R]+ can also be formed stereoselectively. The use of a chiral ionizing medium adds a new dimension, since enantiomers can be distinguished, or even independently identified. The stereochemistry of even-electron cations in the gas-phase is most pronounced with polyfunctional species. The stereochemical behaviour is ruled by two reactivity principles, i.e. the geometry-dependent stabilization of [M + H]+ ions by chelation, and the anchimeric assistance by neighbouring groups in elimination of small molecules (water, ammonia, alcohols, acetic acid, etc.). The stereochemistry of odd-electron cations seems to be governed by three principles, i.e. the thermochemistry of decompositions proceeding with simple-bond cleavage, stereoelectronic effects on bond dissociations in the presence of a control orbital, and long-range interactions resulting in transfer of a hydrogen atom or a larger group. All these three reaction classes have limited areas of application. The stereochemistry of even-electron anions has been developing rapidly. The reactivity of gas-phase anions finds numerous analogies in their chemistry in solution, e.g. hydride transfer reactions and nucleophilic substitution. The applications of mass spectrometry to configurational assignment and structure elucidation remain restricted to selected classes of organic compounds.

An improved method of reconstitution of adipocyte glucose transport activity

The glucose transport activity of rat epididymal fat cells was reconstituted into egg lecithin liposomes with a high degree of reproducibility. The activity was solubilized with 20 m m sodium cholate in Buffer B (10 m m Tris-HCl, pH 7.5). After elimination of small molecules by gel filtration, the transport activity was incorporated into egg lecithin liposomes (Sigma, Type IX-E, homogeneously dispersed into Buffer B) by sonication (5 s), freezing (−70°C), thawing, and a second sonication (5 s). The sonication was done in a 16.8-mm polystyrene test tube (Sarstedt, 55-468) placed in a cup horn (from Heat Systems Ultrasonics) connected to a Branson's sonicator (W-185) at setting No. 3 (70 W of output). The optimum sample size was 80 μl, and the optimum clearance between the test tube and the sonicator horn was 2–3 mm. The concentration of egg lecithin at the reconstitution step was 25 mg/ml, and that of the microsomal protein was approximately 0.3–0.5 mg/ml. The glucose transport activity of reconstituted liposomes was assayed by incubating the latter with a mixture of d-[ 3H]glucose and l-[ 14C]glucose. The incubation was terminated by the addition of HgCl 2, and the reaction mixture was filtered with a Millipore filter (GSWP). The difference in the rates of uptake of d-glucose and l-glucose was regarded as representing the carrier-mediated glucose transport activity. The results of the assay indicated that the glucose transport activity could be reconstituted in a highly reproducible manner. The reconstituted activity was proportional, within a limit of experimental error, to the amount of protein used for reconstitution and was almost completely blocked by cytochalasin B, phloretin, or HgCl 2. However, a small amount of d-glucose was found to bind with the egg lecithin preparation.