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A simple method was developed for the rapid determination of [14C]phosphatidylcholine transfer from small unilamellar liposomes to human plasma HDL, based on the selective precipitation of liposomes by heparin and MnCl2. The assay was utilized to monitor the progress in the partial purification of a phospholipid transfer factor from human plasma. The purification procedure included ultracentrifugation at d = 1.25 g/ml, hydrophobic chromatography on phenyl-Sepharose, affinity chromatography on heparin-Sepharose and gel filtration. The partially purified protein(s) catalyzed the net transfer of phospholipid from small unilamellar phosphatidylcholine liposomes to isolated HDL. The transfer of [14C]phosphatidylcholine from liposomes consisting of phosphatidylcholine/phosphatidylserine/cholesterol (molar ratio, 4:1:5) to HDL was stimulated without affecting the permeability barrier of the liposomal membranes and is, therefore, taken to represent exchange with HDL phospholipid rather than net transfer.
The reaction enthalpies related to the individual steps of two phenolic antioxidants action mechanisms, single electron transfer-proton transfer (SET-PT) and sequential proton loss electron transfer (SPLET), for 30 meta and para-substituted phenols (ArOH) were calculated using DFT/B3LYP method. These mechanisms represent the alternative ways to the extensively studied hydrogen atom transfer (HAT) mechanism. Except the comparison of calculated reaction enthalpies with available experimental and/or theoretical values, obtained enthalpies were correlated with Hammett constants. We have found that electron-donating substituents induce the rise in the enthalpy of proton dissociation (PDE) from ArOH+* radical cation (second step in SET-PT) and in the proton affinities of phenoxide ions ArO- (reaction enthalpy of the first step in SPLET). Electron-withdrawing groups cause the increase in the reaction enthalpies of the processes where electron is abstracted, i.e., in the ionization potentials of ArOH (first step in SET-PT) and in the enthalpy of electron transfer from ArO- (second step in SPLET). Found results indicate that all dependences of reaction enthalpies on Hammett constants of the substituents are linear. The calculations of liquid-phase reaction enthalpies for several para-substituted phenols indicate that found trends hold also in water, although substituent effects are weaker. From the thermodynamic point of view, entering SPLET mechanism represents the most probable process in water.
The streptococcal plasmid pMV158 encodes a gene cassette involved in its mobilization by large conjugative plasmids. Two elements compose this region: i) the mobM gene, encoding the MobM protein that initiates transfer, and ii) the origin of transfer, oriT, which is the target of MobM. In vitro, MobM protein introduces a specific nick within the pMV158-oriT region on supercoiled pMV158 DNA. This paper reports the uncoupling of the oriT and the mobM gene, the latter being placed under the control of an inducible promoter. Upon induction, the vector containing pMV158-oriT was transferred in Escherichia coli matings at a moderate frequency whereas, in vitro, purified MobM protein efficiently cleaved the vector harbouring the pMV158-oriT. Transfer of this vector, as well as transfer of pMV158 in E. coli, required the presence of either the IncW R388 or the IncP RP4 plasmids as auxiliary plasmids. Dissection of the functions encoded by RP4 showed that the traG and traF genes were essential for pMV158 mobilization.
We report collisional energy transfer, CET, quantities for polyatomic-polyatomic collisions and use excited benzene collisions with cold benzene bath, B-B, as our sample system and compare our results with the CET of excited benzene with Ar bath. We find that the gateway mode for both systems is the out-of-plane modes and that in B-B CET, vibration to vibration, V-V, is the dominant channel. Rotations play a mechanistic role in the CET but the net rotational energy transfer is small compared to V-V. The shape of the down side of the energy transfer probability density function, P(E,E'), is convex for B-B collisions and it becomes less so as the temperature increases. In Ar-B collisions, P(E,E') is concave and it becomes less so as the temperature decreases. We report average vibrational, rotational, and translational energy transferred, , as function of temperature for various initial conditions.
This retrospective study is based on 23 males and one female, of an average age of 36.2 years that presented to us between 1982 and 2000 with an average follow up of 61 months, with fully established paralysis of wrist and fingers extension. Fourteen patients had isolated radial nerve palsy, while ten patients had brachial plexus lesions. 1) The tendon transfer for radial nerve palsy was: PT to ECRB, FCU to ED + EPL and PL to APL + EPB; 2) for brachial plexus injury, the tendon transfer was: PT (n = 4) or FDS III or IV (n = 5) to ECRB, FCU (n = 8) or FDS IV (n = 1) to ED + EPL, PL to APL + EPB and wrist arthrodesis with transfer of FDS IV to ED + EPL and PL to APL + EPB. The results were evaluated according to the degree of wrist movement, MP extension of long fingers, opening of first commissure, thumb opposition, grip power and the subjective evaluation of results. Concerning the radial nerve palsy: results are excellent in nine cases and good in one case. An active extension of the wrist of 38 degrees was obtained as well as MP extension of 0 degree with the wrist straightened. Thumb oppositioned was conserved (Kapandji = 8.2), opening of the first commissure 40 degrees and grip power was 20 kg. Concerning the brachial plexus lesions: results are excellent in five cases and good in the other five. An active wrist extension of 32 degrees was obtained, as well as MP extension deficit of 16 degrees with wrist straightened. Opposition was concerned (Kapandji = 7.2), opening of first commissure of 38 degrees and grip power of 13 kg. The functional results are satisfactory, but the analytic study shows some effect of tenodesis of MP extension.
The thiamin diphosphate (ThDP)- and flavin adenine dinucleotide (FAD)-dependent pyruvate oxidase from Lactobacillus plantarum catalyses the conversion of pyruvate, inorganic phosphate, and oxygen to acetyl-phosphate, carbon dioxide, and hydrogen peroxide. Central to the catalytic sequence, two reducing equivalents are transferred from the resonant carbanion/enamine forms of alpha-hydroxyethyl-ThDP to the adjacent flavin cofactor over a distance of approximately 7 A, followed by the phosphorolysis of the thereby formed acetyl-ThDP. Pre-steady-state and steady-state kinetics using time-resolved spectroscopy and a 1H NMR-based intermediate analysis indicate that both processes are kinetically coupled. In the presence of phosphate, intercofactor electron-transfer (ET) proceeds with an apparent first-order rate constant of 78 s(-1) and is kinetically gated by the preceding formation of the tetrahedral substrate-ThDP adduct 2-lactyl-ThDP and its decarboxylation. No transient flavin radicals are detectable in the reductive half-reaction. In contrast, when phosphate is absent, ET occurs in two discrete steps with apparent rate constants of 81 and 3 s(-1) and transient formation of a flavin semiquinone/hydroxyethyl-ThDP radical pair. Temperature dependence analysis according to the Marcus theory identifies the second step, the slow radical decay to be a true ET reaction. The redox potentials of the FAD(ox)/FAD(sq) (E1 = -37 mV) and FAD(sq)/FAD(red) (E2 = -87 mV) redox couples in the absence and presence of phosphate are identical. Both the Marcus analysis and fluorescence resonance energy-transfer studies using the fluorescent N3'-pyridyl-ThDP indicate the same cofactor distance in the presence or absence of phosphate. We deduce that the exclusive 10(2)-10(3)-fold rate enhancement of the second ET step is rather due to the nucleophilic attack of phosphate on the kinetically stabilized hydroxyethyl-ThDP radical resulting in a low-potential anion radical adduct than phosphate in a docking site being part of a through-bonded ET pathway in a stepwise mechanism of ET and phosphorolysis. Thus, LpPOX would constitute the first example of a radical-based phosphorolysis mechanism in biochemistry.
A total of 110 consecutive women was studied prospectively at the time of transcervical embryo transfer following conventional in-vitro fertilization and intracytoplasmic sperm injection procedures. Microbiological cultures were performed on endocervical swabs and embryo transfer catheter tips. Positive microbial growths were observed from endocervical swabs in 78 (70.9%) women and from catheter tips in 54 (49.1%) women. The clinical pregnancy rates were 57.1% in the group of patients without growth and 29.6% in the group with positive microbial growth from catheter tips. As microbial contamination at embryo transfer may influence implantation rates, prospective studies are justified to determine whether eradication of endocervical micro-organisms is possible and whether their eradication will improve implantation rates.
A series of platinum(II) terpyridyl alkynyl complexes, [Pt{4'-(4-R1-C6H4)terpy}(C[triple chemical bond]C-C6H4-R(2)-4)]ClO4 (terpy=2,2':6',2''-terpyridyl; R1=R2=N(CH3)2 (1); R1=N(CH3)2, R2=N-[15]monoazacrown-5 (2); R1=CH3, R2=N(CH3)2 (3); R1=N(CH3)2, R2=H (4); R1=CH3, R2=H (5)), has been synthesized and the photophysical properties of the complexes have been examined through measurement of their UV/Vis absorption spectra, photoluminescence spectra, and transient absorptions. Complex 3 shows a lowest-energy absorption corresponding to a ligand-to-ligand charge-transfer (LLCT) transition from the acetylide to the terpyridyl ligand, whereas 4 shows an intraligand charge-transfer (ILCT) transition from the pi orbital of the 4'-phenyl group to the pi* orbital of the terpyridyl. Upon protonation of the amino groups in 3 and 4, their lowest-energy excited states are switched to dpi(Pt)-->pi*(terpy) metal-to-ligand charge-transfer (MLCT) states. The lowest-energy absorption for 1 and 2 may be attributed to an LLCT transition from the acetylide to the terpyridyl. Upon addition of an acid to a solution of 1 or 2, the amino group on the acetylide is protonated first, followed by the amino group on the terpyridyl. Thus, the lowest excited state of 1 and 2 can be successively switched from the LLCT state to the ILCT state and then to the MLCT state by controlling the amount of the acid added. Such switches in the excited state are fully reversible upon subsequent addition of a base to the solution. Sequential addition of alkali metal or alkaline earth metal ions and then an acid to a solution of 2 also leads to switching of its lowest excited state from the LLCT state, first to the ILCT state and then to the MLCT state. All of the complexes exhibit a transient absorption of the terpyridyl anion radical, which is present in all of the LLCT, ILCT, and MLCT states. However, the shape of the transient absorption spectrum depends on both the substitution pattern on the terpyridyl moiety and the nature of the excited state.
The oxo-Mo(IV) complexes LMoO(S(2)PR(2)-S,S') [L = hydrotris(3,5-dimethylpyrazol-1-yl)borate; R = Me, Et, Pr(i)(), Ph] were prepared by reacting MoO(S(2)PR(2))(2) and KL in refluxing toluene. The dioxo-Mo(VI) complexes cis-LMoO(2)(S(2)PR(2)-S) (R = Pr(i)(), Ph) were prepared by oxidation of the oxo-Mo(IV) complexes or by reaction of LMoO(2)Cl with NaS(2)PR(2). Oxygen atom transfers from Me(2)SO to LMoO(S(2)PR(2)) were first-order with respect to Me(2)SO and complex; the overall second-order rate constants at 40 degrees C range from 9.0(1) x 10(-)(5) M(-)(1).s(-)(1) for LMoO(S(2)PMe(2)) to 2.08(5) x 10(-)(4) M(-)(1).s(-)(1) for LMoO(S(2)PPr(2)); activation parameters were in the ranges DeltaH() = 63(1) to 73(1) kJ.mol(-)(1), DeltaS() = -88(1) to -111(1) J.K(-)(1).mol(-)(1), and DeltaG() = 100(2) kJ.mol(-)(1) for LMoO(S(2)PMe(2)) to 98(2) kJ.mol(-)(1) for LMoO(S(2)PPr(2)). Oxygen atom transfer from pyridine N-oxide to LMoO(S(2)PPr(2)) was also second-order with a rate constant of 1.54(5) x 10(-)(3) M(-)(1).s(-)(1) at 40 degrees C, DeltaH() = 62(1) kJ.mol(-)(1), DeltaS() = -90(1) J.K(-)(1).mol(-)(1), and DeltaG() = 90(1) kJ.mol(-)(1). The second-order rate laws and large negative entropies of activation are consistent with associative mechanisms for the above reactions. Oxygen atom transfer from LMoO(2)(S(2)PPr(2)) to PPh(3) was first-order with respect to reactants, with an overall second-order rate constant of 2.5(3) x 10(-)(4) M(-)(1).s(-)(1) at 30 degrees C. In toluene at 40 degrees C, all the above complexes catalyzed the oxidation of PPh(3) by Me(2)SO, with turnover rates of ca. 0.9 mol of PPh(3)/(mol of catalyst/h). Reduction of LMoO(2)(S(2)PR(2)) by SH(-) led to the generation of the dioxo-Mo(V) anions [LMoO(2)(S(2)PR(2)-S)](-), which were slowly converted to the analogous oxothio-Mo(V) complexes [LMoOS(S(2)PR(2)-S)](-). Dioxygen reacted with [LMoOS(S(2)PPr(2))](-) to produce the oxothio-Mo(VI) complex LMoOS(S(2)PPr(2)-S). The (hydroxo)oxo-Mo(V) complexes LMoO(OH)(S(2)PR(2)-S) were formed upon reduction of LMoO(2)(S(2)PR(2)) with PPh(3) in wet (3-5 M H(2)O) tetrahydrofuran or upon ferrocenium oxidation of LMoO(S(2)PR(2)) in wet tetrahydrofuran. In dry solvents, LMoO(S(2)PR(2)) were oxidized to the corresponding cations, [LMoO(S(2)PR(2)-S,S')](+), which reacted with water to form LMoO(OH)(S(2)PR(2)). The Mo(V) complexes have been characterized by EPR spectroscopy.
The birth of the first baby following in-vitro fertilisation and embryo transfer (IVF/ET) in 1978 and the introduction of gamete intrafallopian transfer (GIFT) in 1984 have increased the treatment modalities in operative reproductive medicine. In tubal pathology, there are besides micro-surgery now so-called additive methods available for treating infertility. With regard to the indications, there has been severe confusion, and it is therefore imperative to define special indications for IVF/ET, microsurgery and GIFT. We do not consider these treatment modalities as alternative methods. Reproductive centres should offer all these treatments to guarantee an appropriate individual programme for each couple. Therefore, knowledge of pregnancy rates is a prerequisite for the doctor. The aim of this paper is, to define the optimal therapy while taking into account the individual problems of each couple.
The full-length cDNA of a phospholipid transfer protein (PLTP) was isolated from Aspergillus oryzae by a RACE-PCR procedure using degenerated primer pool selected from the N-terminal sequence of the purified phosphatidylinositol/phosphatidylglycerol transfer protein (PG/PI-TP). The cDNA encodes a 173 amino acid protein of 18823 Da. The deduced amino acid sequence from position 38 to 67 is 100% identical to the N-terminal sequence (first 30 amino acids) of the purified PG/PI-TP. This amino acid sequence is preceded by a leader peptide of 37 amino acids which is predicted to be composed of a signal peptide of 21 amino acids followed by an extra-sequence of 16 amino acids, or a membrane anchor protein signal (amino acid 5-29). This strongly suggests that the PG/PI-TP is a targeted protein. The deduced mature protein is 138 amino acids long with a predicted molecular mass of 14933 Da. Comparison of the deduced PG/PI-TP sequence with other polypeptide sequences available in databases revealed a homology with a protein deduced from an open reading frame coding for an unknown protein in Saccharomyces cerevisiae (36% identity and 57% similarity). Apart from this homology, the PG/PI-TP is unique and specific to the filamentous fungi on the basis of comparison of PLTP protein sequences. Northern blot analysis of RNA isolated from A. oryzae cultures grown on glucose or glucose supplemented with phospholipids suggests that the PG/PI-TP is transcribed by only one RNA species and allows us to show that expression of the protein is regulated at the messenger RNA level.
Spontaneous revertants of a temperature-sensitive Escherichia coli strain harboring a thermolabile valyl-transfer ribonucleic acid (tRNA) synthetase were selected for growth at 40 degrees C. Of these, a large number still contain the thermolabile valyl-tRNA synthetase. Three of these revertants contained an increased level of the thermolabile enzyme. The genetic locus, valX, responsible for the enzyme overproduction, is adjacent to the structural gene, valS, of valyl-tRNA synthetase. Determination (by radioimmunoassay) of the turnover rates of valyl-tRNA synthetase showed that the increased level of valyl-tRNA synthetase is due to new enzyme synthesis rather than decreased rates of protein degradation.
Plasma cholesteryl ester transfer protein (CETP) concentrations were measured in Japanese subjects by an ELISA with two different monoclonal antibodies that were raised against rabbit CETP and cross-reacted against human CETP. Among 63 patients who consecutively underwent coronary angiography, the plasma CETP of 37 patients with luminal stenosis > or = 50% in their coronary arteries was not significantly different from that of the 26 patients with luminal stenosis < 50%. No other lipoprotein-related measurement except HDL-cholesterol differentiated the two groups. Among 40 hypercholesterolemic patients, no lipoprotein-related measurement other than LDL-cholesterol was found to positive correlate with the CETP. Before and after the treatment of 23 patients with simvastatin 5 mg a day for 4 weeks, plasma CETP markedly decreased in those whose pretreatment CETP was > or = 3 mg/L; no change was observed for those with lower pretreatment CETP. In the former group, negative correlation between CETP and HDL-cholesterol was demonstrated only in the posttreatment plasma.
Two lipid transfer proteins, designated lipid transfer protein-I (Mr 69 000) and lipid transfer protein-II (Mr 55 000), each of which facilitates the transfer of radiolabelled cholesteryl ester, triacylglycerol and phosphatidylcholine between plasma lipoproteins, were purified from human plasma. Immunoglobulin G was prepared from goat antiserum to human lipid transfer protein-I (i.e., anti-human LTP-I IgG). The progressive addition of anti-human LTP-I IgG to buffered solutions containing either a highly purified mixture of human lipid transfer protein-I and lipid transfer protein-II, or highly purified rabbit lipid transfer protein (Abbey, M., Calvert, G.D. and Barter, P.J. (1984) Biochim. Biophys. Acta 793, 471-480) resulted in specific immunoprecipitation and the removal of increasing amounts, up to 100%, of cholesteryl ester, triacylglycerol and phosphatidylcholine transfer activities. However, similar precipitation studies on human and rabbit lipoprotein-free plasma resulted in the progressive removal of all cholesteryl ester and triacylglycerol transfer activities but only 30% (human) or 20% (rabbit) of phosphatidylcholine transfer activity. In all cases more anti-human LTP-I IgG was required to precipitate rabbit lipid transfer activity than human lipid transfer activity. These results suggest that lipid transfer protein-I and lipid transfer protein-II have antigenic sites in common, allowing precipitation of both proteins by specific antibody to lipid transfer protein-I. Most plasma phosphatidylcholine transfer activity is mediated by a protein (or proteins) other than lipid transfer protein-I and lipid transfer protein-II. In lipoprotein-free plasma all cholesteryl ester and triacylglycerol transfer activity, and some phosphatidylcholine transfer activity, is mediated by lipid transfer protein-I (or lipid transfer protein-I and an antigenically similar protein, lipid transfer protein-II.
In a previous study we demonstrated that highly purified lipid-transfer protein facilitated the transfer of triglyceride, cholesteryl ester, and phosphatidylcholine between plasma lipoproteins. It remained unclear, however, whether these lipids were transferred by independent sites on the lipid-transfer protein. To address this point, we have studied the protein-mediated transfer of triglyceride, cholesteryl ester, and phosphatidylcholine as a function of the concentration and lipid composition of donor and acceptor lipoproteins. Lipoproteins labeled in vitro, reconstituted lipoproteins of defined lipid composition, and phosphatidylcholine liposomes with or without triglyceride and/or cholesteryl ester have been used to investigate the inter-relationships of lipids transferred by the lipid-transfer protein. In studies of initial (less than or equal to 10-13%) transfer, we found that, although absolute transfer rates were affected, the ratio of cholesteryl ester to triglyceride transferred was independent of donor and acceptor lipoprotein concentrations and acceptor lipoprotein lipid composition. With reconstituted lipoproteins as donor, we demonstrated that this ratio was linearly related to the ratio of cholesteryl ester to triglyceride in the donor particle; the sum of triglyceride and cholesteryl ester transferred remained constant and independent of the lipid composition of the donor. Experiments with intact lipoproteins labeled in vitro and with small unilamellar vesicles in the presence and absence of p-chloromercuriphenylsulfonate, confirmed the interdependence of triglyceride and cholesteryl ester transfer. In contrast, under all assay conditions, no correlation was found between the amount of phosphatidylcholine transferred and the transfer of triglyceride and/or cholesteryl ester. We conclude that triglyceride and cholesteryl ester compete for transfer and that the extent of transfer for each lipid is determined by its relative concentration in the donor particle, whereas phosphatidylcholine transfer is independent of triglyceride and cholesteryl ester transfer. The data also strongly support the conclusion that lipid transfer protein promotes both the exchange and net transfer of triglyceride and cholesteryl ester and that the net transfer process proceeds by a reciprocal exchange of triglyceride and cholesteryl ester without net transfer of core lipid between lipoproteins.
INTRODUCTION: This study aims to describe the organization that was implemented at the Rhône-Alpes perinatal hotline, as well as to describe in utero transfer and neonate transport from an epidemiological point of view. MATERIAL AND METHOD: A cohort study was performed between January 2003 and December 2004. Every in utero transfer and neonate transport was included. Transfers performed in 2003 were compared to transfers performed in 2004. Three endpoints were defined: the rate of in utero transfer (number of in utero transfers/number of in utero transfers + number of neonatal transfers), the rate of transfer toward level II units (number of transfers from level I to level II/number of transfers from level I to level II + number of transfers from level I to level III) as well as the rate of intra network transfer (number of intra network transfers/number of intra network transfers + number of extra network transfers). RESULTS: In 2003, 865 in utero transfers (IUT) and 1297 neonate transports (NT) were performed, in 2004 848 IUT and 1069 NT were performed. The rate of in utero transfer significantly increased from 40 to 44.2% in 2004 (865/2162 versus 848/1917, p = 0.007). The rate of transfer toward level II units increased for the mothers from 31.8% to 36.9% (177/557 versus 174/471, p = 0.09) and significantly increased for the neonates from 43.2 to 51.6% in 2004 (335/775 versus 327/633, p = 0.002). Finally the rate of intra network transfer has not significantly changed: for the IUT it decreased from 87 to 86% (755/865 versus 732/848, p = 0.59) and for the NT from 91% to 90% (1179/1297 versus 963/1069, p = 0.45). CONCLUSION: The organization that was implemented allows not only a safe 24 hour on call management of maternal transfers as well as neonate transport, but also a precise knowledge of epidemiologic indications relative to perinatal transfer.
OBJECTIVE: To determine the effect of a centralised neonatal transfer service on numbers of neonatal transfers and the time taken for teams to reach the baby. DESIGN: Prospective census of neonatal inter-hospital transfers between May and July 2004. Comparison with a previous census undertaken before introduction of the service. Analysis of requests for antenatal in-utero transfer to the regional emergency bed service. SETTING: Geographically defined area in London and southeast England. PATIENTS: Babies transferred to or from a neonatal unit. INTERVENTIONS: Introduction of a centralised neonatal transfer service. MAIN OUTCOME MEASURES: Numbers of transfers, time taken for teams to arrive to the baby (response time). RESULTS: During the census there were 835 transfers with an increase of 34% from the previous census (n = 619). Most of the increase was in urgent transfers for neonatal intensive care. There was a mean of 4.4 urgent transfers a day, with 3.9 elective and 0.8 short-term transfers. Over the same period in-utero transfers decreased. Response times improved from a median of 2 h in 2001 to 1.45 h in 2004 (p<0.05). The 90th centile fell from 6 h to 4.9 h. CONCLUSION: Following the introduction of a centralised neonatal transfer service, response times improved significantly. An increase in the numbers of transfers for medical intensive care was associated with a reduced number of in-utero transfers. To balance the improved safety and accessibility of neonatal transfer, similar developments may be needed to facilitate in-utero transfer.