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Influenza virus hemagglutinin HA-2 N-terminal fusogenic peptides augment gene transfer by transferrin-polylysine-DNA complexes: toward a synthetic virus-like gene-transfer vehicle.

Complexes containing plasmid DNA, transferrin-polylysine conjugates, and polylysine-conjugated peptides derived from the N-terminal sequence of the influenza virus hemagglutinin subunit HA-2 have been used for the transfer of luciferase or beta-galactosidase marker genes to K562 cells, HeLa cells, and BNL CL.2 hepatocytes. These DNA complexes mimic the entry of viruses into cells, as they contain functions for (i) the packaging of the nucleic acid with polylysine, (ii) the attachment to the cell and receptor-mediated endocytosis with transferrin as a ligand, and (iii) the release from endosomes by using membrane-disrupting influenza peptides. The presence of these influenza peptide conjugates in the DNA complexes renders the complexes active in membrane disruption in a liposome leakage assay and results in a substantial augmentation of the transferrin-polylysine-mediated gene transfer.

Amino Acid Sequence↗

Transfer of specific unresponsiveness to organ allografts by thymocytes. Specific unresponsiveness by thymocyte transfer.

Prolonged survival of vascularized organ allografts has been produced in unmodified inbred rats by transfer of thymocytes from enhanced, engrafted, syngeneic animals. For these thymocytes to increase significantly the survival of test allografts they must be harvested 6-9 d after transplantation. Thymectomy of the enhanced, engrafted animals during the same critical period causes acute rejection of othewise long surviving grafts. For optimal effect, the enhanced thymocyte donor must be actively and passively immunized and receive a cardiac allograft. The necessity for erythrocytes in the initial active immunization regimen is noted. Additionally, the antigenic specificity of the suppressor effect has been established with two histoincompatible donor rat strains. Cellular and humoral host responses mounted by test graft recipients after thymocyte transfer from enhanced, engrafted donors are different from those mounted either by unmodifed animals acutely rejecting their grafts or by enhanced rats bearing well-functioning grafts. Numbers of T lymphocytes are reduced in the grafted hearts and in the spleens of test graft recipients, a finding paralleled by the complete absence of specific direct lymphocyte-mediated cytotoxicity. In contrast, cytotoxic antibody production, although delayed, is increased in magnitude, peaking around the time of graft rejection. These studies provide evidence that different biological manipulations can modify separate pathways in the complex cellular and humoral responses towards organ allografts. They demonstrate that cellular immunity is critically involved in immunological enhancement of vascularized organ allografts, a phenomenon hitherto considered primarily humoral. It seems clear that cells with suppressor activity are present within the thymus during the early phases of immunological enhancement.

Animals↗

Passive transfer of the idiotypically suppressed state by serum from suppressed mice and transfer of suppression from mothers to offspring.

Mice that are suppressed with respect to an idiotype (CRIA) present in A/J anti-p-azophenylarsonate antibodies, hyperimmunized, and allowed to rest were previously found to possess high concentrations of suppressor T cells with anti-idiotypic receptors. We have now observed that the sera of such mice contain soluble factors that can selectively suppress the CRIA component of a humoral response when passively transferred to adult or neonatal recipients. When T cells from suppressed, hyperimmunized mice were transferred into female mice before mating, their offspring, upon immunization, produced anti-Ar antibodies that lacked CRIA. A state of idiotypic suppression was also produced in offspring when the mother was inoculated with serum from suppressed mice a few days before parturition. The results indicate that the suppressor factor is not an immunoglobulin.

Animals↗

Recent horizontal transfer of mellifera subfamily mariner transposons into insect lineages representing four different orders shows that selection acts only during horizontal transfer.

We report the isolation and sequencing of genomic copies of mariner transposons involved in recent horizontal transfers into the genomes of the European earwig, Forficula auricularia; the European honey bee, Apis mellifera; the Mediterranean fruit fly, Ceratitis capitata; and a blister beetle, Epicauta funebris, insects from four different orders. These elements are in the mellifera subfamily and are the second documented example of full-length mariner elements involved in this kind of phenomenon. We applied maximum likelihood methods to the coding sequences and determined that the copies in each genome were evolving neutrally, whereas reconstructed ancestral coding sequences appeared to be under selection, which strengthens our previous hypothesis that the primary selective constraint on mariner sequence evolution is the act of horizontal transfer between genomes.

Amino Acid Sequence↗

Recombination between directly repeated origins of conjugative transfer cloned in M13 bacteriophage DNA models ligation of the transferred plasmid strand.

When two, directly-repeated copies of the origin of transfer (oriT) of the conjugatively mobilizable, broad host-range plasmid R1162 are cloned into bacteriophage M13mp9 DNA, they undergo recombination in the presence of one of the R1162-encoded proteins required for mobilization [Meyer, R. (1989) J. Bacteriol., 171, 799-806]. Mutations in the outer arm of the inverted repeat within oriT inhibit this recombination. These mutations also affect a late step in transfer. We propose that recombination on the phage DNA models the processing of single-stranded DNA after entry into a recipient cell. The two, directly-repeated oriTs are not equivalent during the recombination reaction, because they are differently affected by the outer-arm mutations. A mutation was also isolated that reduces the specificity of the cleavage site in one of the two oriTs. Together, the results with the mutations suggest that phage recombinants can form only when the first cleavage occurs at one of the two oriTs. This is followed by the resulting free 3' end joining to the 5' end at the cleavage site of the other oriT.

Bacteriophages↗

Variation in transfer factors for stochastic models: soil-to-plant transfer.

The scientific literature is a good source of information about soil-to-plant transfer, usually documented as mean concentration ratios. It does not provide, however, good information about variation in concentration ratio values, because most studies are done with limited numbers of crops and soils. Fortunately, a suitably large and diverse database is available from the International Union of Radioecologists that can be used to investigate variation. It has a total of over 7,000 concentration ratio values for over 22 elements. This paper reports on analysis of the geometric standard deviations obtained from the database. We assert that geometric standard deviation values are comparable and transferable among elements, so that geometric standard deviations calculated from large databases can be applied to other elements when there is a paucity of data. For a fully generic situation, where neither crops nor soil types are known, the geometric standard deviations is typically about 6. This implies a 1,300-fold range to encompass 95% of the data. For concentration ratios obtained for crops of interest on a specific site, the geometric standard deviation values are much lower, about 1.5. This still implies a fivefold range to encompass 95% of the data. Other values of geometric standard deviations for intermediate levels of information are provided. These estimates of geometric standard deviation are important to and appropriate for stochastic simulations of the impacts of soil contamination on crops and subsequent food chains. They are also important to the inclusivity of concentration ratios value distributions for unusual but perhaps important crops and settings.

Agriculture↗

A comparison of temporalis transfer and free latissimus dorsi transfer in lower facial reanimation following unilateral longstanding facial palsy.

The ultimate goal in the treatment of facial palsy is the restoration of voluntary and spontaneous movement to the paralyzed side of the face, symmetrical to the normal side. We report our experience treating 40 patients with established facial palsy over a 4-year period. All patients underwent either temporalis transfer or free latissimus dorsi transfer as a single stage to improve lower facial symmetry. We believe that both techniques reliably achieve an increase of movement in facial reanimation after oncological, traumatic, or congenital facial palsy.

Adolescent↗

Transfer of Kingella indologenes (Snell and Lapage 1976) to the genus Suttonella gen. nov. as Suttonella indologenes comb. nov.; transfer of Bacteroides nodosus (Beveridge 1941) to the genus Dichelobacter gen. nov. as Dichelobacter nodosus comb. nov.; and assignment of the genera Cardiobacterium, Dichelobacter, and Suttonella to Cardiobacteriaceae fam. nov. in the gamma division of Proteobacteria on the basis of 16S rRNA sequence comparisons.

The 16S rRNA sequences of Kingella indologenes, Cardiobacterium hominis, and Bacteroides nodosus were determined by direct RNA sequencing, using a modified Sanger method. Sequence comparisons indicated that these three species represent a novel family in the gamma division of Proteobacteria. On the basis of these data, K. indologenes and B. nodosus cannot retain their current generic status as they are not closely related to other members of their assigned genera. Therefore, we propose transfer of K. indologenes to the new genus Suttonella as Suttonella indologenes and transfer of B. nodosus to the new genus Dichelobacter as Dichelobacter nodosus and assign the genera Cardiobacterium, Suttonella, and Dichelobacter to a new family, Cardiobacteriaceae, in the gamma division of Proteobacteria.

Bacteroides↗

Proton-transfer and non-transfer in compounds of quinoline and quinaldic acid with L-tartaric acid.

The structures of two compounds of L-tartaric acid with quinoline, viz. the proton-transfer compound quinolinium hydrogen (2R,3R)-tartrate monohydrate, C9H8N+.C4H5O6-.H2O, (I), and the anhydrous non-proton-transfer adduct with quinaldic acid, bis(quinolinium-2-carboxylate) (2R,3R)-tartaric acid, 2C10H7NO2.C4H6O6, (II), have been determined at 130 K. Compound (I) has a three-dimensional honeycomb substructure formed from head-to-tail hydrogen-bonded hydrogen tartrate anions and water molecules. The stacks of pi-bonded quinolinium cations are accommodated within the channels and are hydrogen bonded to it peripherally. Compound (II) has a two-dimensional network structure based on pseudo-centrosymmetric head-to-tail hydrogen-bonded cyclic dimers comprising zwitterionic quinaldic acid species which are interlinked by tartaric acid molecules.

Journal Article↗

Thermodynamics of information transfer between subunits in oligomeric enzymes and kinetic cooperativity. 3. Information transfer between the subunits of chloroplast fructose bisphosphatase.

The theory and the methods that have been described in the two preceding papers in this journal have been used to analyze the kinetic properties of chloroplast fructose bisphosphatase. The enzyme is a tetramer made up of apparently identical subunits and displays a sigmoidal kinetics with respect to its substrate, fructose bisphosphate. The free ionic species, magnesium and fructose bisphosphate bind to the enzyme and the chelate fructose-bisphosphate-magnesium does not affect the sigmoidicity of the rate curves. The Hill coefficient with respect to free fructose bisphosphate is equal to 2.3, which is indeed incompatible with the view that the enzyme behaves as a dimer of dimers. This conclusion is confirmed by direct analysis of the rate curve. On the basis of the sum of the residuals, their sum of squares, the standard error of the kinetic parameters of the equation, the kinetic scheme associated with a dimer of dimers may be ruled out. On the basis of the same criteria, the fit of an Adair equation to the rate data cannot be retained as satisfactory. This is a direct proof that neither the Monod nor the Koshland model can correctly fit these kinetic data. In fact the model that fits these data best is a structural kinetic scheme where information transfer occurs between each subunit and its three neighbors ('tetrahedral' mode of information transfer). The fit of these models to a large number of kinetic data allows one to compute the free energy profile during the successive binding processes of the four substrate molecules to the enzyme. Whereas the first two steps are associated with an increase of free energy, all the other subsequent steps are associated with a decrease of free energy.

Chloroplasts↗

Three cases of delivery under sodium valproate--placental transfer, milk transfer and probable teratogenicity of sodium valproate.

Here is a report of three cases of delivery from women who have taken sodium valproate (VPA). One neonate was born with polydactylism, and we reviewed the congenital malformations in children of mothers who have taken VPA. In the other two cases, we also examined the levels of VPA in the cord serum and breast milk. The levels of VPA in the cord serum were 50 micrograms/ml at 12 hours after the last maternal dose and 75 micrograms/ml at 14 hours after the last maternal dose. The blood levels of VPA in the two mothers were 48-59 micrograms/ml and 55-85 micrograms/ml in their late pregnancy, respectively. The levels of VPA in the breast milk were 2.0-3.5 micrograms/ml and the ratio of VPA in the breast milk to blood was 2-3%. These results suggest the high rate of placental transfer and the low rate of milk transfer of VPA.

Abnormalities, Drug-Induced↗

Passive transfer of streptozotocin induced diabetes mellitus with spleen cells. Studies of synogeneic and allogeneic transfer to normal and athymic nude mice.

This study shows passive transfer of streptozotocin induced diabetes mellitus in mice. Transplants of spleen cells from BALB/c mice, streptozotocin treated for 5 days, induced diabetes in normal BALB/c recipients. Treatment of transplants with anti-theta and complement resulted in a significant decrease in the degree of diabetes. Athymic nude mouse recipients of BALB/c background also developed diabetes after transpant of spleen cells from both syngenic and allogenic (C-7/Bl/6) donors. It is concluded that passive transfer of chemically induced diabetes in mice is practicable, in both syngeneic and allogeneic combinations, and that thymus derived lymphocytes are significant in this process.

Animals↗

Chlorophyll photosensitized electron transfer reactions in lipid vesicles: enhancement in yield of vectorial electron transfer across the bilayer from reduced cytochrome c to oxidized ferredoxin by addition of valinomycin plus potassium ion.

Chlorophyll photosensitized electron transfer across a vesicle bilayer from reduced cytochrome c in the inner compartment to oxidized ferredoxin in the outer compartment, using propylene diquat as a mediator, has been investigated using both steady-state and laser flash photolysis methods. One of the factors limiting the quantum yield is the transmembrane potential, which is formed during sample preparation and is increased by the electron transfer process across the membrane bilayer. This limitation can be diminished by the incorporation of valinomycin into the bilayer in the presence of potassium ion. The overall quantum yield can be approximately doubled (up to a total of 22% based on the chlorophyll triplet which is quenched, and 2.8% based on the absorbed quanta) by valinomycin addition. Another quantum yield limitation arises from the accumulation of oxidized cytochrome c in the inner aqueous compartment, which is formed as a consequence of the transbilayer electron transport process and can quench triplet chlorophyll on the inner side of the vesicle. The chlorophyll cation radical generated in this way can participate in the electron exchange equilibrium between chlorophyll molecules located within the bilayer, and thus inhibit electron flow from inside to outside. This acts to limit the extent of cytochrome c oxidation to less than or equal to 50% of the original amount.

Chlorophyll↗

Control of Interspecies Electron Flow during Anaerobic Digestion: Significance of Formate Transfer versus Hydrogen Transfer during Syntrophic Methanogenesis in Flocs.

Microbial formate production and consumption during syntrophic conversion of ethanol or lactate to methane was examined in purified flocs and digestor contents obtained from a whey-processing digestor. Formate production by digestor contents or purified digestor flocs was dependent on CO(2) and either ethanol or lactate but not H(2) gas as an electron donor. During syntrophic methanogenesis, flocs were the primary site for formate production via ethanol-dependent CO(2) reduction, with a formate production rate and methanogenic turnover constant of 660 muM/h and 0.044/min, respectively. Floc preparations accumulated fourfold-higher levels of formate (40 muM) than digestor contents, and the free flora was the primary site for formate cleavage to CO(2) and H(2) (90 muM formate per h). Inhibition of methanogenesis by CHCl(3) resulted in formate accumulation and suppression of syntrophic ethanol oxidation. H(2) gas was an insignificant intermediary metabolite of syntrophic ethanol conversion by flocs, and its exogenous addition neither stimulated methanogenesis nor inhibited the initial rate of ethanol oxidation. These results demonstrated that >90% of the syntrophic ethanol conversion to methane by mixed cultures containing primarily Desulfovibrio vulgaris and Methanobacterium formicicum was mediated via interspecies formate transfer and that <10% was mediated via interspecies H(2) transfer. The results are discussed in relation to biochemical thermodynamics. A model is presented which describes the dynamics of a bicarbonate-formate electron shuttle mechanism for control of carbon and electron flow during syntrophic methanogenesis and provides a novel mechanism for energy conservation by syntrophic acetogens.

Journal Article↗

Beginning a genetic analysis of conjugational transfer determined by the F factor in Escherichia coli by isolation and characterization of transfer-deficient mutants.

Eighty-four transfer-deficient mutants of Flac have been isolated; 27 of these bear amber mutations and 1 mutant is temperature-sensitive. All the mutants transfer between 10(-2) and <10(-5)% as well as wild-type Flac, all are curable by acridine orange treatment, and all are resistant to the female-specific phage phi(II). Some of the mutants are partially sensitive to female-specific phage tau. Sixty-three of the mutants are resistant to the male-specific phages f1, f2, and Qbeta; 15 are resistant only to f2; and 6 are sensitive to all three male-specific phages. Most of the mutants are still poor recipients in conjugation, but four of the mutants resistant to f1, f2, and Qbeta have become good recipients. Those mutants resistant to all three male-specific phages do not seem to make F-pili.

Acridines↗

Use of the oxygen transfer slope and estimated membrane oxygen transfer to predict PaO2.

The purpose of this investigation was threefold: (1) could the perfusionist accurately estimate oxygen transfer (VO2/minute) while on CPB; (2) could this estimate, and its position on the oxygen transfer slope (OTS), predict resultant PaO2 values within a specific range; and (3) could previously derived performance 'normals' be used during this study. Fifty-four sets of samples (both arterial and venous) from 27 oxygenators were used in this study. Each oxygenator provided one normothermic and one hypothermic set of samples. In 48 of the 54 samples (88.9%) the predicted VO2/minute was within +/- 10% of the actual VO2/minute. Thirty-nine of these 48 (81.25%) had resultant PaO2 values within our target range of 140 +/- 30 mmHg. The PaO2 for these 39 samples ranged from 110 to 168 mmHg with a mean of 133 mmHg. The percentage of predicted shunt (POPS) ranged from 59.0 to 192.4% with a mean of 109.3% (SD = 23.81%). With this degree of variability, we concluded that the perfusionist must assess VO2/minute as well as POPS in order to predict the resultant PaO2.

Arteries↗

Transference--counter-transference constellations and enactments in the psychotherapy of destructive narcissism.

Destructive narcissism, characterized by a perverse alliance between the self and an omnipotent bad internal object, constitutes a serious obstacle to psychoanalytic psychotherapy. This paper draws together various object relations theories of destructive narcissism, and presents a case study of a failed psychotherapy to illustrate the transference--counter-transference constellations and their associated enactments in treatment of this disturbance.

Countertransference↗