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[Culture media for in vitro fertilization].

The potential of a medium used for IVF-ET must be defined not only by the percentages of successful fertilization and cleavage, but above all by the percentage of term pregnancies. However, they can only be expressed in a well-defined environmental context. The medium has to remain in equilibrium with its gas phase, which can modify the biophysical parameters (pH, pO2, redox potential), and can even be deleterious (presence of NO and CO). Several factors can also interfere with the results obtained in theoretically identical media. In powder media, a certain lack, of homogeneity and an inferior quality of the water added can be major sources of variability. The definitions of the actual compositions of the media can vary continuously and insidiously if the original datum is not consistently taken as reference. These two points are developed in the paper. A great number of facts have, nevertheless, been established: the optimal biophysical conditions, the mineral composition, the amino acid composition, and the necessity for certain substrates such as pyruvate. Numerous lacunae remain: vitamin requirements, the supply of lipids and macromolecular compounds. The addition of serum to the culture media has not helped to remove these uncertainties. We have recently shown that serum is not indispensable in the IVF-ET medium. This opens the way to an understanding of the mechanisms of in vitro fertilization and early embryonic development.

Amino Acids↗

Glucocorticoid receptors and glucocorticoid sensitivity of human leukemic cells.

We have established optimal conditions for the measurement of glucocorticoid receptors (GR) in human white cells using a whole-cell binding assay with [3H]dexamethasone as the ligand, and the subsequent determination of the GR content in normal human lymphocytes and in leukemic cells of patients with various forms of acute and chronic leukemia. A number of leukemia cell lines in continuous culture were also subjected to the GR assay, and the results were correlated with the sensitivity of these cell lines to glucocorticoid steroids in vitro. The GR content of normal human lymphocytes amounted to 4,850 +/- 1,340 (mean +/- SD) receptors/cell. The mean equilibrium dissociation constant (KD) of the interaction of [3H]dexamethasone with the GR was 1.2 x 10(-8) M. Steroidal compounds with a known glucocorticoid potency effectively competed for the binding, whereas steroids devoid of glucocorticoid activity (e.g. estradiol-17 beta and testosterone) were ineffective. The GR content of the blast cells obtained from eight patients suffering from acute leukemia and four patients with a blast crisis of chronic myelocytic leukemia was found to be highly variable (3,230-29,900 receptors/cell), while the lymphocytes of six patients with chronic lymphatic leukemia contained a rather stable GR content (2,930-5,120 receptors/cell), which was comparable with that of normal lymphocytes. GR was identified in all the 12 malignant continuous white cell lines studied. Large cells contained more GR than the smaller ones. There was no apparent correlation between the GR concentration and the sensitivity of the cells in vitro to glucocorticoids as judged by [3H]thymidine incorporation studies. Distribution of the surface markers in the leukemic cell lines did not relate to the GR concentration. We conclude that the presence of GR is probably a universal feature of the leukemic cells, and, from a clinical standpoint, probably does not alone imply steroid responsiveness.

Adolescent↗

A continuous fluorometric assay for phospholipases using polymerized mixed liposomes.

A versatile continuous fluorometric assay for phospholipases A2, C, and D has been developed utilizing polymerized mixed liposomes made of pyrene-containing phospholipids (5 mol%) uniformly inserted in the polymerized liposomes of 1,2-bis[12-(lipoyloxy)dodecanoyl]-sn-glycero-3-phosphoglycerol (BLPG) and its derivatives. 1-Hexadecanoyl-2-(1-pyrenedecanoyl)-sn-glycero-3-phosphocholine was used for phospholipase A2 and N-(1-pyrenesulfonyl)-egg phosphatidyl ethanolamine for phospholipases C and D. Fluorescence emission of pyrene moieties in polymerized mixed liposomes was strongly quenched by BLPG molecules and, thus, the hydrolysis of pyrene-containing phospholipids and the subsequent displacement of pyrene moieties from the liposomes resulted in a large increase in fluorescence intensity. All the phospholipases tested selectively and rapidly hydrolyzed the inserted pyrene-containing phospholipids, which were readily monitored by measuring an increase in fluorescence emission at 380 nm. Assay conditions for individual phospholipases were optimized by altering interfacial properties of polymerized liposomes, such as surface charge, and subsequently by changing the chemical structure of hydrolyzable phospholipids. Phospholipase activities were linearly proportional to enzyme concentrations in the range from 0.1 to 50 ng. Specific activity determined for phospholipases from a wide variety of sources ranged from 0.5 to 100 mumol/min/mg. Polymerized mixed liposomes are exceptionally stable against chemical and physical degradation and the assay requires only a small amount of pyrene-containing phospholipids. In addition, the polymerized matrix of BLPG (and its derivatives), due to its inertness to the phospholipase hydrolysis, allows the direct measurement of the equilibrium dissociation constant for a protein-liposome complex.

Fluorometry↗

Comparison of the membrane-related effects of cytarabine and other agents on model membranes.

UNLABELLED: The membrane-associated effects of a series of chemotherapeutic and other drugs were examined via differential scanning calorimetry and by their modulation of the action of porcine phospholipase A2 (PLA2) on bilayer substrates. The drugs examined included: cytarabine, amino-glycoside antibiotics, adriamycin, dibucaine, butacaine, and VP-16. The bilayers employed were phase-separated ternary lipid mixtures containing dimyristoylphosphatidylcholine: palmitoyllysolecithin: and either hexadecanoic acid (fatty acid ternary mixture) or hexadecanol (alcohol ternary mixture). Effects of the more hydrophilic drugs (cytarabine and aminoglycoside antibiotics) on the calorimetric profiles of the negatively charged (fatty acid-containing) and the neutral (hexadecanol-containing) ternary lipid mixtures indicate that the interaction of these drugs with biomembranes is likely to be dominated by electrostatic interactions. All of the drugs investigated, including the more hydrophobic adriamycin, dibucaine, butacaine, and VP-16, affected the phase equilibrium in the membrane and exhibited apparent noncompetitive inhibition of the action of PLA2 on bilayers composed of ternary lipid substrates. In addition, cytarabine inhibited fusion of fatty acid-containing ternary mixtures. CONCLUSIONS: These drug:membrane interactions leading to a shift in the phase equilibria were apparently regiospecific. Hydrophilic drug:membrane interactions included an important electrostatic component. The effects of all of the drugs employed in this study on the action of PLA2 on a bilayer substrate (fatty acid-containing ternary lipid mixture) are hypothesized to be a result of the drug-mediated shift in phase equilibria away from the optimally active phase distribution. As a result, PLA2 binds with normal affinity to the membrane, but its membrane substrate is not catalytically turned over. It is evident that these drugs can directly affect cellular homeostasis in a manner that can show a dependence on the nature of the membrane surface.

Anti-Bacterial Agents↗

Single-step formation of a biorecognition layer for assaying histidine-tagged proteins.

The purpose of this work was to develop a simple procedure for the creation of a specific biorecognition layer for histidine-tagged (His-tagged) molecules. Such a layer was prepared by the spontaneous fusion of vesicles containing readily available plain (DOPC) and iminodiacetic acid (DOGS-NTA) phospholipids on a silica surface resulting in the formation of an NTA-containing supported lipid bilayer. The frequency surface acoustic waveguide device which supports Love waves was used to follow the real-time formation of the biorecognition layer. The mole percent of the DOGS-NTA phospholipids in the supported bilayer was optimized by following the kinetics of the fusion for the different NTA-containing lipids. Fluorescently labeled lipids were used with observations of the fluorescence recovery after photobleaching to confirm the presence of lipid bilayers. After saturating all NTA-molecules with Ni(2+), the binding of a His-tagged protein fragment within the concentration range of 0.04 and 0.4 mM to a 5 mol % DOGS-NTA/DOPC was detected; binding curves were used to calculate the apparent association constant k(on) = 2.56 x 10(4) M(-)(1) s(-)(1), dissociation constant k(off) = 1.3 x 10(-)(3) s(-)(1), and equilibrium constant k(eq) = 1.97 x 10(7) M(-)(1). The described method could find significant applications as a generic technique for preparing biorecognition layers for His-tagged proteins. In addition, the acoustic waveguide device, which provides high sensitivity together with flexibility in terms of the substrate material used, is shown to be an attractive alternative to direct optical biosensors.

Choline↗

Prion peptide fragment PrP[106-126] forms distinct cation channel types.

Using the lipid bilayer technique we have optimized the recording conditions and confirmed that PrP[106-126] (KTNMKHMAGAAAAGAVVGGLG) forms single ion channels. Based on the conductance and kinetic parameters of the single channel currents recorded in 250/50 mM KCl cis/trans we have found that the PrP[106-126]-formed heterogeneous cation channels that differ in their conductance and kinetic properties. The most frequently observed PrP[106-126]-formed single cation channels were those of: (a) a GSSH- and TEA-sensitive channel with fast kinetics (n = 47). The current-voltage (I-V) relationship of this channel, that has a reversal potential E(rev) of -33 mV close to the equilibrium potential for K(+) (E(K) -35 mV), exhibited inward and outward rectification. The values of the maximal slope conductance (g(max)) were 138 and 141 pS at positive and negative potentials, respectively. The values of the permeability ratios were 1.0:0.87:0.72:0.49:0.41 for K(+) > Rb(+) > Na(+) > Cs(+) > Li(+) respectively. The probability of the channel being open (P(o)) and the frequency (F(o)) had bell-shaped curves with a peak at membrane potential (V(m)) between -10 and -5 mV whereas the mean open and closed times (T(o) and T(c)) had inverted bell-shaped curves; (b) a 4'-4'-dithiodipyridin (DTT)-sensitive channel with slow kinetics (n = 32). The I-V relationship of this channel that had an E(rev) of -35 mV and a g(max) of 41 pS at positive V(m) was non-linear. The parameter P(o) increased at positive V(m) to 0.6-0.7 at +80 mV. F(o) had an asymmetrical bell-shaped curve with a peak of 314 events/sec at -80 mV. The values of T(o) and T(c) were 312 and 164 msec at +120 mV, respectively; (c) a large channel (n = 24 channels) that had five equally spaced subconductances showed time-dependent fast and slow transitions at positive and negative V(m), respectively. The inactivation ratio I(ss)/I(i) was V(m) dependent and described by a bell-shape. The I-V relationship of this channel that had a E(rev) of -22 mV was non linear. The value of g(max) was 900 and 1444 pS at positive and negative V(m)s, respectively. The value of P(o) was 0.6 at negative V(m)s between -160 and -80 mV and 0.23 at +140 mV. F(o) increased from 22 events/sec at -160 mV to 80-100 events/sec at between +80 and +100 mV. T(o) decreased from 375 msec between -160 and -80 mV to 1-2 msec at V(m)s between 0 and +160 mV. In contrast, T(c) decreased from 160-240 msec at membrane voltages (V(m)s) between -160 and -80 mV. The biophysical properties of these channels indicate that they are capable of modifying cellular functions via modification of V(m) and electrolyte homeostasis of the cell.

Cations, Monovalent↗

The influence of binding to albumin and alpha 1-acid glycoprotein on the clearance of drugs by the liver.

The liver is a major site for synthesis and catabolism of plasma proteins. Albumin has various binding sites for anionic drugs, alpha 1-acid glycoprotein possesses a single binding site for cationic drugs. In spite of extensive protein binding, the liver can efficiently remove drugs from the circulation. Intrahepatic dissociation of the drug-protein complex may involve dissociation-limited debinding under non-equilibrium conditions or surface interaction-facilitated dissociation phenomena. During liver or renal disease and acute-phase conditions plasma protein binding of drugs may be affected. Changes in the unbound drug fraction do not always result in proportional changes in clearance or distribution volume. Potential changes in the unbound concentration in steady-state as well as the fluctuations in total plasma levels depend on the extent of protein binding of a drug, the relative change in the unbound drug fraction type of clearance, the size of the distribution volume, route of administration as well as concomitant changes in intrinsic (cellular) clearance function. Optimization of dosage regimens for certain drugs and interpretation of liver function tests with diagnostic dyes may largely benefit from determination of the unbound rather than the total concentration of the drugs involved.

Humans↗

A DFT investigation of conformational geometries and interconversion equilibria of phenylthiosemicarbazone and its complexation with zinc.

The geometrical structures of phenylthiosemicarbazone (HAPhTSC) conformers have been obtained by geometry optimizations using density functional theory (DFT) calculations at the B3LYP/6-31G(d) and B3LYP/6-311G(d,p) levels of theory. Six thioamino and 24 thioimino tautomers of HAPhTSC have been found. Six tautomerization reactions between thioamino and thioimino tautomers occurring via transition states and their corresponding activation energies have been obtained. Conformational pathways for tautomerizations and interconversions of HAPhTSC conformers have been presented. Tautomerization between the most stable species of thioamino (Atttcc) and its thioimino (Itttcct) tautomer is an endothermic reaction, Delta H(0)=18.17 kcal mol(-1) and its log K=-13.74, at 298.15 K. Thermodynamic quantities of tautomerizations, interconversions of HAPhTSC conformers and their equilibrium constants are reported. The geometry of the zinc complex with HAPhTSC, found as a Zn(HAPhTSC)2Cl2 structure, has been obtained using B3LYP/6-31G(d) calculations. Binding of the Zn(HAPhTSC)2Cl2 complex is an exothermic and spontaneous reaction. [figure]. Conformational notation defined as a name consisting of a letter "A" for A thioamino tautomer followed by "c" for cis or "t" for trans isomerism of five dihedral angles of chi(C4-C3-C2-N3), phi(C3-C2-N3-N2), psi(C2-N3-N2-C1), theta;(N3-N2-C1-N1) and omega(N2-C1-N1-H2), serially, or a letter "I" for B thioimino tautomer followed by "c" for cis or "t" for trans isomerism of six dihedral angles of chi(C4-C3-C2-N3), phi(C3-C2-N3-N2), psi(C2-N3-N2-C1), theta(N3-N2-C1-N1), omega(N2-C1-N1-H2) and delta (N2-C1-S-H1), serially.

Computational Biology↗

Verification of tangential breast treatment dose calculations in a commercial 3D treatment planning system.

The accuracy of the photon convolution/superposition dose algorithm employed in a commercial radiation treatment planning system was evaluated for conditions simulating tangential breast treatment. A breast phantom was fabricated from machineable wax and placed on the chest wall of an anthropomorphic phantom. Radiographic film was used to measure the dose distribution at the axial midplane of the breast phantom. Subsequently, thermoluminescent dosimeters (TLDs) were used to measure the dose at four points within the midplane to validate the accuracy of the film dosimetry. Film measurements were compared with calculations performed using the treatment planning system for four types of treatment: optimized wedged beams at 6 and 18 MV and two-dimensional compensated beams at 6 and 18 MV. Both the film- and TLD-measured doses had a precision of approximately 0.6%. The film-measured doses were approximately 1.5% lower than the TLD-measured doses, ranging from 0-3% at 6 MV and 0.5-1% at 18 MV. Such results placed a high level of confidence in the accuracy and precision of the film data. The measured and calculated doses agreed to within +/-3% for both the film and TLD measurements throughout the midplane exclusive of areas not having charged particle equilibrium. Good agreement was not expected within these regions due to the limitations in both film dosimetry and the dose-calculation algorithm. These results indicated that the treatment planning system calculates doses at the midplane with clinically acceptable accuracy in conditions simulating tangential breast treatment.

Algorithms↗

Quantification of D2-like dopamine receptors in the human brain with 18F-desmethoxyfallypride.

UNLABELLED: Substituted benzamides such as (11)C-raclopride or (123)I-iodobenzamide are selective radiotracers for PET and SPECT imaging of D(2)-like dopamine (DA) receptors. (18)F-Desmethoxyfallypride ((18)F-DMFP) is a benzamide tracer with the advantage of an (18)F label. We optimized the synthesis and evaluated (18)F-DMFP in PET studies on healthy human volunteers. METHODS: The affinity of DMFP for D(2)-like DA receptors was characterized in vitro using membrane preparations from rat striatum and the DA receptor ligand (3)H-spiperone. PET studies on 10 healthy human volunteers were performed using a whole-body PET scanner after injection of 214 +/- 54 MBq (mean +/- SD) (18)F-DMFP. Brain images were acquired dynamically over 124 min, and metabolite-corrected plasma activity was used as the input function. Data analysis was performed using several different approaches (compartmental, graphical, equilibrium methods). RESULTS: The mean inhibition constant (K(i)) of DMFP was 15 +/- 9 nmol/L. In human brain, the striatum-to-cerebellum ratio reached a maximum of about 4 between 60 and 120 min. When specific binding in the striatum was expressed as the difference between binding in the striatum and the cerebellum, it reached a maximum at approximately 60 min after injection and remained almost constant until the end of data acquisition. The ratio of specific striatal to nonspecific cerebellar binding was about 3:1 at 120 min after injection. A small, but significant specific tracer binding could also be detected in the thalamus. Treatment of a schizophrenic patient with a high dose (1,000 mg/d) of another substituted benzamide, amisulpride, resulted in a reduction of specific tracer uptake of about 90% in striatal regions. With regard to measured distribution volumes and binding potentials, there was an excellent agreement between all applied analytic methods. CONCLUSION: Our study demonstrates that (18)F-DMFP is a highly reliable tracer for PET imaging of D(2)-like DA receptors. It offers the major advantage that it can be used independently of an on-site cyclotron within a PET satellite network. Noninvasive analytic methods without blood sampling provide valid measurements of receptor quantities in human striatum. Because of the (18)F label and the favorable imaging properties, (18)F-DMFP could become an efficient substitute for (11)C-raclopride in a clinical context.

Adult↗

Characterization of a thromboxane A2/prostaglandin H2 receptor in guinea pig lung membranes using a radioiodinated thromboxane mimetic.

Thromboxane A2 (TXA2) and prostaglandin H2 (PGH2) are potent constrictors of airway smooth muscle and may mediate some of the pulmonary effects of leukotrienes. To date, the TXA2/PGH2 receptor in lung has not been well characterized. In this report, we describe the evaluation of the TXA2/PGH2 receptor in guinea pig lung membranes using the new radiolabeled TXA2 mimetic [1S(1 alpha,2 beta(5Z),3 alpha(1E,3S*),4 alpha)]-7-[3-(3-hydroxy-4-(4'- iodophenoxy)-1-butenyl)-7-oxabicyclo-[2.2.1]heptan-2-yl]-5-h eptenoic acid (IBOP). IBOP elicited a dose-dependent contraction of guinea pig lung parenchymal strips (EC50 = 3.03 +/- 0.97 nM, three experiments), which was blocked by the TXA2/PGH2 antagonists SQ29548 (pKB = 7.44 +/- 0.2, three experiments), BM13505 (pKB = 6.29 +/- 0.26, three experiments), and I-PTA-OH (pKB = 5.82 +/- 0.36, three experiments). In radioligand binding studies, the binding of [125I]IBOP to guinea pig lung membranes prepared from perfused lungs was saturable, displaceable, and dependent upon protein concentration. Binding was optimal at pH 6.5 and was enhanced by the addition of mono- and divalent cations. The standard assay buffer was 25 mM 3-(N-morpholino)propanesulfonic acid, pH 6.5, 100 mM NaCl, 5 mM MgCl2. Binding was inhibited by pretreatment with dithiothreitol, N-ethylmaleimide, or beta-mercaptoethanol. Binding was unaffected by the addition of guanine nucleotide analogs at concentrations up to 300 microM. Analysis of the time course of binding of [125]IBOP at 30 degrees yielded k-1 = 0.0447 min-1, k1 = 2.49 x 10(8) M-1 min-1, and Kd = k-1/k1 = 180 pM. Computer analysis of equilibrium binding studies using nonlinear methods (LUNDON-1) revealed a single class of noninteracting binding sites with a Kd of 86.9 +/- 11.9 pM and a Bmax of 81.8 +/- 7.7 fmol/mg of protein (three experiments). [125I]IBOP binding to guinea pig lung membranes was inhibited by a series of TXA2/PGH2 receptor agonists and antagonists, with a rank order different from that previously determined for washed guinea pig platelets (Spearman's r = 0.686, p greater than 0.05). [125I]IBOP binding to guinea pig lung membranes was also inhibited by the prostanoids prostaglandin D2, prostaglandin E2, prostaglandin F2 alpha, and 9 alpha,11 beta-prostaglandin F2, all of which have been proposed to act at the TXA2/PGH2 receptor in lung.

Animals↗

Metal bioremediation through growing cells.

Heavy-metal pollution represents an important environmental problem due to the toxic effects of metals, and their accumulation throughout the food chain leads to serious ecological and health problems. Metal remediation through common physico-chemical techniques is expensive and unsuitable in case of voluminous effluents containing complexing organic matter and low metal contamination. Biotechnological approaches that are designed to cover such niches have, therefore, received great deal of attention in the recent years. Biosorption studies involving low-cost and often dead/pretreated biomass have dominated the literature and, subsequently, extensive reviews focusing on equilibrium and kinetics of metal biosorption have also come up. However, the low binding capacity of biomass for certain recalcitrant metals such as Ni and failure to effectively remove metals from real industrial effluents due to presence of organic or inorganic ligands limit this approach. At times, when pure biosorptive metal removal is not feasible, application of a judicious consortium of growing metal-resistant cells can ensure better removal through a combination of bioprecipitation, biosorption and continuous metabolic uptake of metals after physical adsorption. Such approach may lead to simultaneous removal of toxic metals, organic loads and other inorganic impurities, as well as allow optimization through development of resistant species. However, sensitivity of living cells to extremes of pH or high metal concentration and need to furnish metabolic energy are some of the major constraints of employing growing cells for bioremediation. The efforts to meet such challenges via isolation of metal-resistant bacterial/fungal strains and exploitation of organic wastes as carbon substrates have began. Recent studies show that the strains (bacteria, yeast and fungi) isolated from contaminated sites possess excellent capability of metal scavenging. Some bacterial strains possess high tolerance to various metals and may be potential candidates for their simultaneous removal from wastes. Evidently, the stage has already been set for the application of metal-resistant growing microbial cells for metal harvesting. This review focuses on the applicability of growing bacterial/fungal/algal cells for metal removal and the efforts directed towards cell/process development to make this option technically/economically viable for the comprehensive treatment of metal-rich effluents.

Adsorption↗

Hydration of an amphiphilic excipient, Gelucire 44/14.

The incorporation of drugs into Gelucires has been reported to increase the dissolution rate of poorly soluble drugs, often leading to improved drug bioavailability. In pharmaceutical applications, it is important to know how the excipient interacts with the drug, and how the mixture behaves during manufacturing, storage as well as during administration. The uptake of water by an amphiphilic excipient, Gelucire 44/14, has been investigated in two ways: storage in humid air and addition of liquid water. During exposure to humid air, the uptake goes in stages that correspond to the dissolution of the components of the excipient, starting with the most hydrophilic ones: glycerol, then polyethylene glycol (PEG), PEG esters (PEG monolaurate and PEG dilaurate), and finally glycerides (trilaurin). At each stage, the remaining crystals are in equilibrium with an interstitial solution made of water and the dissolved components. In this range of hydrations, the total uptake is close to the sum of the equilibrium hydrations of the components. In the pharmaceutical formulation, the active ingredient could dissolve in the liquid phase. At larger hydrations, obtained through addition of liquid water, the state of Gelucire 44/14 differs from those of its components. Gelucire 44/14 forms a lamellar phase and this phase melts at 30 degrees C whereas the pure PEG esters form hexagonal and cubic mesophases. The cubic mesophases do not melt until the temperature exceeds 40 degrees C. At body temperature, all crystals in Gelucire 44/14 melt to an isotropic fluid as soon as the total water content exceeds 5%. Therefore the formulation of amphiphilic excipients can be optimized to avoid the formation of mesophases that impede dissolution of the excipient at body temperature.

Body Fluids↗

Iodide modulation of the EDTA-induced iodine reductase activity of horseradish peroxidase by interaction at or near the EDTA-binding site.

Horseradish peroxidase (HRP) catalyses the reduction of iodinium ion (I+) to iodide by H2O2 in the presence of EDTA. I+ reduction occurs optimally at pH 6 whereas the enzyme catalyses iodide oxidation optimally at pH 3.5. Thus the two activities reside on the same enzyme with two characteristic pH optima. Iodide modulates the expression of the reductase activity by EDTA. Higher concentrations of iodide inhibit the reductase activity by EDTA. Nitrite, an electron donor, acts similarly to iodide. Both EDTA and nitrite competitively inhibit iodide oxidation, indicating that they compete with iodide for the same binding site for electron flow to the haem iron group. However, unlike iodide, EDTA converts compound I, not into the native enzyme, but into a compound absorbing at 416 nm which reduces I+ and then returns to the native form. The apparent equilibrium dissociation constant, KD, for the formation of the EDTA-HRP complex (15 mM) is doubled in the presence of iodide, indicating interference with EDTA binding by iodide. EDTA binds away from the haem iron centre and not through intramolecular Ca2+. The pH-dependence of EDTA binding indicates that an ionizable group of the enzyme with pKa 5.8, presumably a distal histidine, controls the binding. The data suggest that iodide competes with EDTA for compound I and modulates the iodine reductase activity by limiting the formation of the 416 nm-absorbing active compound.

Anions↗

Specific external forcing of spatiotemporal dynamics in reaction-diffusion systems.

Self-organization behavior and in particular pattern forming spatiotemporal dynamics play an important role in far from equilibrium chemical and biochemical systems. Specific external forcing and control of self-organizing processes might be of great benefit in various applications ranging from technical systems to modern biomedical research. We demonstrate that in a cellular chemotaxis system modeled by one-dimensional reaction-diffusion equations particular forms of spatiotemporal dynamics can be induced and stabilized by controlling spatially distributed influx patterns of a chemical species as a function of time. In our model study we show that a propagating wave with certain shape and velocity and static symmetrical and asymmetrical patterns can be forced and manipulated by numerically computing open-loop optimal influx controls.

Biophysics↗

Non-equilibrium thermodynamic sensitivity of oxidative phosphorylation.

A new method was developed to analyse the dynamic properties of oxidative phosphorylation, in particular the sensitivity of the phosphate potential with respect to fluctuating cellular ATP utilization. This treatment is based on the eigenvalue sensitivity analysis of an experimentally supported non-equilibrium thermodynamic model of oxidative phosphorylation. Such an analysis allows direct access to the kinetic information, while circumventing the awkward conventional numerical integration of a set of nonlinear differential equations. This procedure revealed, for the parameters characteristic for liver of starved rats in vivo, that the sensitivity of oxidative phosphorylation to a fluctuating ATP utilization is minimal at a degree of coupling q = 0.95. This means that the phosphate potential is highly buffered with respect to fluctuating energy demands at the degree of coupling. This value of q agrees well with the degree of coupling qeef, at which net ATP production of oxidative phosphorylation--at optimal efficiency--occurs in the most economic way. This simultaneous maximization of kinetic stability and economic thermodynamic efficiency at the same degree of coupling appears to be a coincidence.

Adenosine Triphosphate↗

[Assessment of potential blood substitute solutions with oxygen-carrying properties and their possible uses].

Beside the necessary volume effect (isoosmotic and isooncotic solution) and the physiological bicarbonate concentration (prevention of dilution acidosis) a possible blood substitute with O2 carrier characteristics should enable an optimal O2 transport: Viscosity as low as possible and shear rate independent (Newtonian behavior), O2 concentration at the given O2 partial pressure at least 6 ml/dl, and mean capillary O2 partial pressure as high as possible. For the judgement of such a blood substitute, the so-called O2 supply index is recommended, i.e., O2 concentration times mean capillary O2 partial pressure divided by viscosity.

Acid-Base Equilibrium↗

Purification of herpesvirus saimiri and properties of the viral DNA.

Conditions for growth, concentration, and purification of Herpesvirus saimiri were determined. Optimal yields of infectious Herpesvirus saimiri (HVS) were obtained from infected owl monkey kidney (OMK) cells grown at 32.5 degrees C in medium containing 10 per cent fetal calf serum. Forth-five percent of the initial infectious HVS was recovered after an 18-fold concentration using 8 per cent polyethylene glycol 6000 in the presence of 0.5 M NaCl. Polyethylene glycol concentrated HVS was purified in an isopycnic-linear Renografin gradient (1.0-1.3 g/cm3. Ninety-six percent of the infectivity was recovered in a single 1.16 g/cm3 density region. DNA extracted from purified HVS was resolved into two distinct density classes by CsCl equilibrium centrifugation (1.727 and 1.709 g/cm3). DNase treated HVA virions yield four DNA species with densities of 1.727, 1.718, 1.712, and 1.706 g/cm3 in CsCl centrifugation.

Cell Line↗