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PubMed2000

Differential scanning calorimetric and Fourier transform infrared spectroscopic studies of the effects of cholesterol on the thermotropic phase behavior and organization of a homologous series of linear saturated phosphatidylserine bilayer membranes.

We have examined the effects of cholesterol on the thermotropic phase behavior and organization of aqueous dispersions of a homologous series of linear disaturated phosphatidylserines by high-sensitivity differential scanning calorimetry and Fourier transform infrared spectroscopy. We find that the incorporation of increasing quantities of cholesterol progressively reduces the temperature, enthalpy, and cooperativity of the gel-to-liquid-crystalline phase transition of the host phosphatidylserine bilayer, such that a cooperative chain-melting phase transition is completely or almost completely abolished at 50 mol % cholesterol, in contrast to the results of previous studies. We are also unable to detect the presence of a separate anhydrous cholesterol or cholesterol monohydrate phase in our binary mixtures, again in contrast to previous reports. We further show that the magnitude of the reduction in the phase transition temperature induced by cholesterol addition is independent of the hydrocarbon chain length of the phosphatidylserine studied. This result contrasts with our previous results with phosphatidylcholine bilayers, where we found that cholesterol increases or decreases the phase transition temperature in a chain length-dependent manner (1993. Biochemistry, 32:516-522), but is in agreement with our previous results for phosphatidylethanolamine bilayers, where no hydrocarbon chain length-dependent effects were observed (1999. Biochim. Biophys. Acta, 1416:119-234). However, the reduction in the phase transition temperature by cholesterol is of greater magnitude in phosphatidylethanolamine as compared to phosphatidylserine bilayers. We also show that the addition of cholesterol facilitates the formation of the lamellar crystalline phase in phosphatidylserine bilayers, as it does in phosphatidylethanolamine bilayers, whereas the formation of such phases in phosphatidylcholine bilayers is inhibited by the presence of cholesterol. We ascribe the limited miscibility of cholesterol in phosphatidylserine bilayers reported previously to a fractional crystallization of the cholesterol and phospholipid phases during the removal of organic solvent from the binary mixture before the hydration of the sample. In general, the results of our studies to date indicate that the magnitude of the effect of cholesterol on the thermotropic phase behavior of the host phospholipid bilayer, and its miscibility in phospholipid dispersions generally, depend on the strength of the attractive interactions between the polar headgroups and the hydrocarbon chains of the phospholipid molecule, and not on the charge of the polar headgroups per se.

Biophysical Phenomena↗
PubMed2003

A direct comparison of surface and bulk chain-relaxation in polystyrene.

Near-edge X-ray absorption fine-structure (NEXAFS) spectroscopy was used to measure simultaneously the relaxation rates of polystyrene (PS) molecules at the free surface and in the bulk. The samples were uniaxially stretched and annealed at temperatures below the bulk glass transition temperature of PS. The surface and bulk chain relaxation was monitored by measuring the partial-electron and the fluorescence NEXAFS yields, respectively, both parallel and perpendicular to the stretching direction. The decay of the optical birefringence was also measured to provide an independent measure of the bulk relaxation. Relaxation of PS chains was found to occur faster on the surface relative to the bulk. The magnitude of the surface glass transition temperature suppression over the bulk was estimated based on the information on the temperature dependence of the rates.

Journal Article↗
PubMed2003

Direct imaging of nanoparticle embedding to probe viscoelasticity of polymer surfaces.

Atomic force microscopy was used to study the embedding of gold nanoparticles into the surface of polystyrene films. The rate of embedding was determined at temperatures near the bulk glass transition temperature T(g) by measuring the apparent nanosphere height as a function of annealing time. In particular, relative height measurements of the adhered particles were made at temperatures below the bulk T(g) value. In the absence of enhanced surface dynamics or yield processes no embedding is expected to occur for T<T(g). Measurements on 10 and 20 nm particles both indicated that the particles did embed 3-4 nm into the polymer for T<T(g). Both the extent and time frame for engulfment appear to be independent of the particle diameter. The results suggest a more mobile surface region on the order of 3-4 nm thick, with a lower glass transition temperature than the bulk.

Journal Article↗
PubMed1988

Temperature-dependent osmotic permeability in glycoprotein containing liposomes.

The osmotic water outflow of large multilamellar liposomes containing alpha 1-acid glycoprotein was measured at a temperature near the lipid's phase transition temperature. The liposomes were formed from a mixture of DPPC, cholesterol and glycoprotein in molar ratios 100:20:1, by continuous sucrose density gradient centrifugation. These liposomes captured 35% of the radiolabeled glycoprotein. The temperature-dependent experiments showed that near phase transition temperature the initial rate of water outflow increased drastically in comparison with glycoprotein free liposomes incubated in buffer containing glycoprotein. We suggested that eventual a channel mechanism may be involved due to spontaneous incorporation of glycoprotein into the bilayer.

In Vitro Techniques↗
PubMed2005

Application of thermally responsive polypeptides directed against c-Myc transcriptional function for cancer therapy.

Elastin-like polypeptides are biopolymers composed of the pentapeptide repeat Val-Pro-Gly-Xaa-Gly. Elastin-like polypeptides are soluble in aqueous solution below their transition temperature, but they hydrophobically collapse and aggregate when the temperature is raised above the transition temperature. Previous studies have suggested that the aggregation of these polypeptides in response to externally applied hyperthermia may be exploited in the use of elastin-like polypeptide for thermally targeted drug delivery. This work shows the application of elastin-like polypeptide as a delivery vehicle for a short peptide that can inhibit the transcriptional function of a specific oncogene. The coding sequence for elastin-like polypeptide was modified by the addition of the membrane translocating sequence penetratin and a peptide derived from helix 1 of the helix-loop-helix region of c-Myc (H1-S6A,F8A), known to inhibit c-Myc transcriptional function. The designed polypeptide (Pen-ELP-H1) was then expressed and purified from Escherichia coli. Cellular uptake of Pen-ELP-H1 is enhanced by both the penetratin sequence and by the hyperthermia-induced phase transition as shown by flow cytometry studies. Using immunofluorescence and reverse transcription-PCR, we show that Pen-ELP-H1 is able to disrupt the nuclear localization of c-Myc and inhibit transcriptional activation by c-Myc. Cell proliferation studies showed that Pen-ELP-H1 inhibits growth of MCF-7 cells. Furthermore, the use of hyperthermia increased the antiproliferative effect of a thermally responsive Pen-ELP-H1 approximately 2-fold compared with a nonthermally responsive control polypeptide. These studies show that genetically engineered elastin-like polypeptide carriers may provide a new way to thermally target specific oncogene inhibitors to solid tumors.

Amino Acid Sequence↗
PubMed2006

Sterilization of gentamicin containing collagen/PLGA microparticle composites.

In order to achieve implants which provide sustained release of gentamicin, microparticles based on a 50/50 Resomer 503/Resomer 502H blend were combined with collagen in order to achieve their fixation and to utilize the favorable effect of collagen on wound healing. Ethylene oxide treatment as well as beta- and gamma-irradiation were tested for sterilization of the collagen/PLGA-microparticle composite. All methods resulted in a decrease of molecular weight and glass transition temperature of polymer raw material and microparticles. In addition, ethylene oxide treatment yielded aggregation of microparticles leading to a substantial increase in the initially liberated gentamicin dose. Furthermore, chemical changes of gentamicin after ethylene oxide sterilization could be identified using NMR spectroscopy. Despite a decrease in the molecular weight and glass transition temperature after irradiation, neither morphological changes of the composites nor changes regarding the gentamicin release profile from beta- and gamma-sterilized material were observed. Free radicals, which could only be detected in gentamicin drug substance and at marginal level in gentamicin-loaded MPs, disappeared within 4 weeks. Additional microbiological testing verified the microbiological activity of gentamicin liberated from beta-sterilized composites. Storage of beta-sterilized composite at 4 degrees C/35% r.h. for 3 months did not influence morphology, molecular weight, glass transition temperature, and release profiles of microparticles and composites. However, at 25 degrees C/60% r.h. and 40 degrees C/75% r.h. a marked decrease in molecular weight and glass transition temperature resulted. This effect was due to a higher humidity, water uptake into polymers, and subsequent hydrolysis of polymers and microparticles, which was more pronounced for RG 502H because of its hydrophilicity. Upon storage at 25 degrees C/60% r.h. and 40 degrees C/75% r.h. particles collapsed resulting in an increased gentamicin liberation. Thus, all sterilization techniques have their pros and cons, but based on drug release profile and chemical changes of gentamicin irradiation treatment appears to be more suitable for collagen/gentamicin-loaded PLGA microparticle composites.

Beta Particles↗
PubMed2005

Interaction of tetraphenyl-porphyrin derivatives with DPPC-liposomes: an EPR study.

The effect of the symmetry and polarity of the porphyrin molecules on their membrane localization and interaction with membrane lipids were investigated by electron paramagnetic resonance (EPR). For this purpose, two glycoconjugated tetraphenyl porphyrin derivatives were selected, respectively, symmetrically and asymmetrically substituted. Small unilamellar liposomes composed of dipalmitoylphosphatidylcholine (DPPC) and spin labeled stearic acids were prepared. The spin probe was located at the 5th or 7th or 12th or 16th position of the hydrocarbon chain in order to monitor various regions of the lipid bilayer. EPR spectra of porphyrin-free and porphyrin-bound liposomes were recorded at various temperatures below and above the phase transition temperature of DPPC. The effect on membrane fluidity proved to be stronger with the asymmetrical porphyrin derivative than with the symmetrical one. The rigidity increased when the spin label was near lipid head groups. The difference observed between control and porphyrin-treated samples when measured below the main lipid transition temperature disappeared at higher temperature. When the spin label was near the end of the hydrophobic tails, the symmetrical porphyrin derivative caused increase in fluidity, while the asymmetrical one slightly decreased it. To explain this phenomenon we propose that the asymmetrical derivative exerts a stronger ordering effect caused by its fluorophenyl group located at the level of the lipid heads, which is attenuated to the hydrophobic tails. The perturbing effect of the symmetric derivative could not lead to similar extent of ordering at the head groups and looses the hydrocarbon chains deeper in the membrane.

1,2-Dipalmitoylphosphatidylcholine↗
PubMed2006

Observation of stripe mobility in a dipolar frustrated ferromagnet.

We have discovered two novel aspects of the stripe-domain to paramagnetic transition in perpendicularly magnetized Fe films on Cu(100). First, the width of the stripes carrying oppositely oriented spins decreases, close to the transition temperature, with a power law. Second, in a small temperature interval close to the transition temperature, the stripes--which form stationary patterns at low temperatures--become mobile. Various theoretical works have predicted stripe mobility in similar frustrated systems but no direct proof of this phenomenon has been reported so far.

Journal Article↗
PubMed1993

Lipid concentration affects the kinetic stability of dielaidoylphosphatidylethanolamine bilayers.

The bilayer to hexagonal phase transition temperature (Th) of dielaidoylphosphatidylethanolamine is 65.5 degrees C as measured by DSC heating scans at lipid concentrations below 100 mg/ml and at scan rates ranging from 1.7 to 45 degrees C/h. However, at lipid concentrations above 100 mg/ml and at scan rates of 1-3 degrees C/h the measured Th decreases below 65.5 degrees C. At a lipid concentration of 500 mg/ml and a heating scan of 1.2 degrees C/h the transition to the hexagonal phase occurs at 62.7 degrees C. However, this same sample scanned at a rate of 34 degrees C/h has a transition temperature of 64.6 degrees C. Thus a combination of high lipid concentration and slow scan rate is required to significantly lower the hexagonal phase transition temperature below 65 degrees C. These results demonstrate that the rate of conversion of the bilayer to the hexagonal phase is dependent on the concentration of the lipid suspension even under conditions of full hydration. Furthermore, a 100 mg/ml suspension of this lipid which has a Th of 64.3 degrees C at a scan rate of 3.2 degrees C/h has a lower hexagonal phase transition temperature of 62.8 degrees C after pelleting the lipid with low-speed centrifugation but retaining the same amount of solvent in the supernatant above the pellet. Pelleting of the lipid also has a marked effect on the isothermal rate of conversion of the bilayer to hexagonal phase as observed by 31P NMR. The conversion is highly temperature-dependent and is orders of magnitude more rapid for the pelleted sample than for the suspension.(ABSTRACT TRUNCATED AT 250 WORDS)

Calorimetry, Differential Scanning↗
PubMed1979

The influence of charge on phosphatidic acid bilayer membranes.

A complete titration of phosphatidic acid bilayer membranes was possible for the first time by the introduction of a new anaologue, 1,2-dihexadecyl-sn-glycerol-3-phosphoric acid, which has the advantage of a high chemical stability at extreme pH values. The synthesis of the phosphatidic acid is described and the phase transition behaviour in aqueous dispersions is compared with that of three ester phosphatidic acids; 1,2-dimyristoyl-sn-glycerol-3-phosphoric acid, 1,3-dimyristoylglycerol-2-phosphoric acid and 1,2-dipalmitoyl-sn-glycerol-3-phosphoric acid. The phase transition temperatures (Tt) of aqueous phosphatidic acid dispersions at different degrees of dissociation were measured using fluorescence spectroscopy and 90 degrees light scattering. The Tt values are comparable to the melting points of the solid phosphatidic acids in the fully protonated states, but large differences exist for the charged states. The Tt vs. pH diagrams of the four phosphatidic acids are quite similar and of a characteristic shape. Increasing ionisation results in a maximum value for the transition temperatures at pH 3.5 (pK1). The regions between the first and the second pK of the phosphatidic acids are characterised by only small variations in the transition temperatures (extended plateau) in spite of the large changes occurring in the surface charge of the membranes. The slope of the plateau is very shallow with increasing ionisation. A further decrease in the H+ concentration results in an abrupt change of the transition temperature. The slope of the Tt vs. pH diagram beyond pK2 becomes very steep. This is the result of reduced hydrocarbon interaction energy, which was demonstrated by differential scanning calorimetry (Blume, A. and Eibl, H., unpublished data).

Fluorescence Polarization↗
PubMed1992

Lateral conductance parallel to membrane surfaces: effects of anesthetics and electrolytes at pre-transition.

The effects of dilute salts and anesthetics were studied on the impedance dispersion in the dipalmitoylphosphatidylcholine (DPPC) liposomes. Below the pre-transition temperature, the apparent activation energy for conductance in DPPC-H2O without salts was equivalent to pure water, 18.2 kJ mol-1. This suggests that the mobile ions (H3O+ and OH-) interact negligibly with the lipid surface below the pre-transition temperature. At pre-transition temperature, the apparent activation energy of the conductance decreased by the increase in the DPPC concentrations. The effects of various salts (LiCl, NaCl, KCl, KBr, and KI) on the apparent activation energy of the conductance were studied. Changes in anions, but not in cations, affected the activation energy. The order of the effect was Cl- less than Br- less than I-. Cations appear to be highly immobilized by hydrogen bonding to the phosphate moiety of DPPC. The smaller the ionic radius, the more ions are fixed on the surface at the expense of the free-moving species. The apparent activation energy of the transfer of ions at the vesicle surface was estimated from the temperature-dependence of the dielectric constant, and was 61.0 kJ mol-1 in the absence of electrolytes. In the presence of electrolytes, the order of the activation energy was F- greater than Cl- greater than Br- greater than I-. When the ionic radius is smaller, these anions interact with the hydration layer at the vesicle surface and the ionic transfer may become sluggish. In the absence of electrolytes, the apparent activation energy of the dielectric constant decreased by the increase in halothane concentrations. In the presence of electrolytes, however, the addition of halothane increased the apparent activation energy. We propose that the adsorption of halothane on the vesicle surface produces two effects: (1) destruction of the hydration shell, and (2) increase in the binding of electrolytes to the vesicle surface. In the absence of electrolytes, the first effect predominates and the apparent activation energy is decreased. In the presence of electrolytes, the latter effect predominates and the apparent activation energy is increased.

1,2-Dipalmitoylphosphatidylcholine↗
PubMed1977

The intermediate monoclinic phase of phosphatidylcholines.

Two pure phospholipids, dimyristoyl phosphatidylcholine and dipalmitoyl phosphatidylcholine, have been studied using freeze-fracture electron microscopy and the partitioning of the spin label, TEMPO. It is found that the characteristic band pattern, corresponding to monoclinic symmetry in multilamellar liposomes, is observed only in freeze-fracture electron microphotographs when samples are quenched from temperatures intermediate between the chain melting transition temperature and the pretransition temperature of the membrane. Markings are also observed on fracture faces of samples quenched from below the pretransition, but these "bands" are few in number and are widely and irregularly spaced. The lipid membranes used for freeze-fracture were prepared using detergent dialysis and are thought to consist of one, two, or some small number of concentric bilayer shells. These observations are in excellent accord with the recent, prior studies of Janiak, M.J., Small, D.M. and Shirley, G.G., ((1976) Biochemistry 15, 4575--4580), who found monoclinic symmetry (Pbeta' structure) in multilamellar liposomes of these phospholipids only when the sample temperature was intermediate between the main, chain melting transition temperature, and the pretransition temperature. The significance of these results for relating freeze-fracture electron microphotographis to phase diagrams derived from spin label or calorimetric data is discussed briefly. 2,2,6,6-Tetramethylpiperidine-1-oxyl (TEMPO) partitioning data show distinct differences between liposomal preparations of these lipids, and other preparations having fewer bilayers per vesicular structure, with respect to the position, width, and hysteresis of the pretransition.

Electron Spin Resonance Spectroscopy↗
PubMed2003

Observation of surface and bulk phase transitions in nematic liquid crystals.

The behaviour of liquid crystal (LC) molecules near a surface is of both fundamental and technological interest: it gives rise to various surface phase-transition and wetting phenomena, and surface-induced ordering of the LC molecules is integral to the operation of LC displays. Here we report the observation of a pure isotropic-nematic (IN) surface phase transition-clearly separated from the bulk IN transition-in a nematic LC on a substrate. Differences in phase behaviour between surface and bulk are expected, but have hitherto proved difficult to distinguish, owing in part to the close proximity of their transition temperatures. We have overcome these difficulties by using a mixture of nematic LCs: small, surface-induced composition variations lead to complete separation of the surface and bulk transitions, which we then study independently as a function of substrate and applied magnetic field. We find the surface IN transition to be of first order on surfaces with a weak anchoring energy and continuous on surfaces with a strong anchoring. We show that the presence of high magnetic fields does not change the surface IN transition temperature, whereas the bulk IN transition temperature increases with field. We attribute this to the interaction energy between the surface and bulk phases, which is tuned by magnetic-field-induced order in the surface-wetting layer.

Journal Article↗
PubMed2007

Modification and optimization of the united-residue (UNRES) potential energy function for canonical simulations. I. Temperature dependence of the effective energy function and tests of the optimization method with single training proteins.

We report the modification and parametrization of the united-residue (UNRES) force field for energy-based protein structure prediction and protein folding simulations. We tested the approach on three training proteins separately: 1E0L (beta), 1GAB (alpha), and 1E0G (alpha + beta). Heretofore, the UNRES force field had been designed and parametrized to locate native-like structures of proteins as global minima of their effective potential energy surfaces, which largely neglected the conformational entropy because decoys composed of only lowest-energy conformations were used to optimize the force field. Recently, we developed a mesoscopic dynamics procedure for UNRES and applied it with success to simulate protein folding pathways. However, the force field turned out to be largely biased toward -helical structures in canonical simulations because the conformational entropy had been neglected in the parametrization. We applied the hierarchical optimization method, developed in our earlier work, to optimize the force field; in this method, the conformational space of a training protein is divided into levels, each corresponding to a certain degree of native-likeness. The levels are ordered according to increasing native-likeness; level 0 corresponds to structures with no native-like elements, and the highest level corresponds to the fully native-like structures. The aim of optimization is to achieve the order of the free energies of levels, decreasing as their native-likeness increases. The procedure is iterative, and decoys of the training protein(s) generated with the energy function parameters of the preceding iteration are used to optimize the force field in a current iteration. We applied the multiplexing replica-exchange molecular dynamics (MREMD) method, recently implemented in UNRES, to generate decoys; with this modification, conformational entropy is taken into account. Moreover, we optimized the free-energy gaps between levels at temperatures corresponding to a predominance of folded or unfolded structures, as well as to structures at the putative folding-transition temperature, changing the sign of the gaps at the transition temperature. This enabled us to obtain force fields characterized by a single peak in the heat capacity at the transition temperature. Furthermore, we introduced temperature dependence to the UNRES force field; this is consistent with the fact that it is a free-energy and not a potential energy function. beta

Biophysics↗
PubMed1978

Phase Behavior of Chloroplast and Microsomal Membranes during Leaf Senescence.

Wide angle x-ray diffraction of chloroplast and microsomal membranes from primary leaves of Phaseolus vulgaris has revealed that for both types of membrane, portions of the lipid become crystalline as the tissue senesces. For young leaves the transition temperature is about 23 C for microsomes and below -30 C for chloroplast membranes, indicating that at physiological temperature the lipid is entirely liquid-crystalline. Between 2 and 3 weeks after planting the transition temperature rises to 38 C for microsomes, but for chloroplasts this increase to a point above physiological temperature does not occur until between 3 and 4 weeks. Thereafter the transition temperature continues to rise for both types of membrane with advancing senescence, although the rate of increase is greater for chloroplasts than for microsomes. The appearance at physiological temperature of gel phase lipid in the microsomes coincides temporally with the initiation of a decline in total protein in the tissue, and the incidence of crystallinity in chloroplasts coincides with loss of chlorophyll. This change in phase behavior cannot be attributed to an alteration in fatty acid composition, but for both membrane systems it correlates with an increase of about 4-fold in the sterol to phospholipid ratio.

Journal Article↗
PubMed2002

[Lanthanide ion luminescence probe: low temperature phase transition of EuS4N complex].

Fluorescence spectrum of EuS4N complex at low temperature is very different from that at room temperature. When temperature changes, shapes of both excitation and emission spectra change dramatically at about 160 K, which is taken as an indicator of the structural change of EuS4N. Peak splitting of 5D0-->7F0 also indicates that there are two different types of coordination structure for Eu3+ ions at low temperature, while only one at room temperature. Thermal effect for the structure transition is also deduced from the relation between fluorescence intensity and temperature changing manner. These results were in good accordance with directly measurement on sample temperature. The reason for the structure change is also discussed from the structure of the EuS4N complex. This study proved that lanthanide luminescent probe, as a complementarity to X-ray diffraction technique, should be an effective tool in low temperature crystal structure study.

Chelating Agents↗
PubMed1977

Lipid crystallization in senescent membranes from cotyledons.

Lipid transition temperatures for rough and smooth microsomal membranes isolated from bean (Phaseolus vulgaris) cotyledon tissue at various stages of germination were determined by wide angle x-ray diffraction. The transition temperatures were established by recording diffraction patterns through a temperature series until a sharp x-ray reflection centered at a Bragg spacing of 4.15 A and denoting the presence of crystalline lipid was discernible. For rough and smooth microsomes from 2-day-old tissue, the transitions occurred at 0 C and 3 C, respectively, indicating that at this early stage in the germination sequence the membrane lipid is entirely liquid-crystalline at physiological temperature. By the 4th day of germination, the transition temperatures had increased to 32 C for smooth microsomes and 35 C for rough microsomes, indicating that at 29 C, which was the growth temperature, portions of the membrane lipid were crystalline. During the later stages of germination, the transition temperature for smooth microsomes continued to rise through 44 C at day 7 to 56 C at day 9, by which time the cotyledons were extensively senescent and beginning to abscise. There was also a dramatic increase in the proportion of membrane lipid in the crystalline phase at 29 C. By contrast, the rough microsomes showed little change in transition temperature and only a slight increase in the proportion of crystalline lipid during this late period in germination. The data indicate that substantial amounts of the lipid is senescing membranes are crystalline even at physiological temperature. Moreover, there is a temporal correlation between the appearance of this crystallinity and loss of membrane function, suggesting that the two may be causally related.

Journal Article↗
PubMed1995

Temperature-controlled high-performance liquid chromatography using a uniformly sized temperature-responsive polymer-based packing material.

Uniformly sized polymer-based packing materials modified with a temperature-responsive polymeric selector, poly(N-isopropylacrylamide), on either the internal or the external surface were prepared by an in situ surface-selective modification method. These packing materials were utilized as stationary phases in reversed-phase high-performance liquid chromatography to evaluate their ability to thermally control chromatographic separation selectivity. A packing material having a modified internal surface showed temperature-dependent separation selectivity in drug separation. Drugs that could not be separated below the phase transition temperature of the poly(N-isopropylacrylamide) selector were separated faster and with much better resolution above the phase transition temperature, while the unmodified base packing material afforded worse resolution of the drugs above the phase transition temperature due to the peak overlapping resulting from faster elution of all the solutes employed. In addition, a packing material modified on the external surface showed a temperature-responsive shift of the elution region for a polypeptide such as BSA, which was thought to be useful in direct injection of a serum sample.

Acrylic Resins↗