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PubMed2003

Indirect imaging of ethanol via magnetization transfer at high and low magnetic fields.

Ethanol (EtOH) is believed to exert its neurochemical effects through interactions with brain cellular components, which causes a fraction of brain EtOH to have a lower molecular mobility. This facilitates magnetization transfer to other molecules similarly associated with macromolecules, such as water. It was hypothesized that this effect can be used in vivo to image EtOH indirectly via the much stronger brain tissue water resonance. EtOH-containing bovine serum albumin samples were used to demonstrate magnetic coupling between EtOH and water at 7 T and 1.5 T. Spectroscopy and imaging experiments demonstrated that EtOH signal saturation yielded greater water signal reduction than inversion and that this reduction scaled with EtOH concentration in the BSA samples. In human brain at physiologically relevant brain EtOH concentrations, water signal reductions were measurable when saturating the EtOH resonance. Strengths and limitations of indirectly imaging brain EtOH are discussed.

Brain↗
PubMedDate not supplied

Evaluation of the accuracy of 3 transfer techniques for implant-supported prostheses with multiple abutments.

PURPOSE: This study evaluated the deformation of a metallic framework connected to 15 stone casts fabricated using 3 transfer techniques to determine the most accurate impression procedure. MATERIALS AND METHODS: Five stone casts were made from polyether impressions of an epoxy resin master model for each transfer technique. Group 1 samples were created by the direct splinted technique (square transfer copings splinted with carbon steel pins and autopolymerizing acrylic resin, custom tray); group 2 samples were made by the direct nonsplinted technique (square transfer copings, custom tray); and group 3 samples were fabricated using the indirect technique (tapered transfer copings, custom tray). Sixteen strain gauges were fixed on the framework to measure the degree of framework deformation for each stone cast. Pairs of strain gauges placed opposite each other constituted 1 channel to read deformation (half Wheatstone bridge). Deformation readings were collected at the 4 segments between abutments in 4 directions (anterior, posterior, superior, and inferior). Deformation data were analyzed using analysis of variance and the Tukey test at the .05 and .01 levels of significance. RESULTS: Group 1 samples allowed the most accurate reproduction of analog position compared to the samples made using the other techniques. No significant difference was found between the direct nonsplinted (group 2) and indirect (group 3) techniques. DISCUSSION: Although some studies have evaluated transfer techniques with similar methodology, this study demonstrated the most suitable strain gauge setup to record framework deformations in all directions and simultaneously offset the effects of temperature variation. CONCLUSIONS: The direct splinted technique was the most accurate transfer method for multiple abutments compared to direct nonsplinted and indirect techniques.

Acrylic Resins↗
PubMed2005

Mechanism of HCS + O2 reaction: hydrogen- or oxygen-transfer?

In spite of the potential importance of the HCS radical in both combustion and interstellar processes, its chemical reactivity has not been tackled previously. In the present paper, the oxidation reaction of the HCS radical is theoretically investigated for the first time at the CCSD(T)/6-311++G(3df,2p)//BH&HLYP/6-311++G(d,p)+ZPVE and Gaussian-3//B3LYP/6-31G(d) levels. It is shown that the most feasible pathway is the O2 addition to the HCS radical forming the intermediate SC(H)OO which can undergo a subsequent O-extrusion leading to SC(H)O + 3O. This features an indirect O-transfer mechanism with the overall barrier of 4.4 and 3.5 kcal mol(-1), respectively, at the two levels. However, formation of the H-transfer product CS + HO2 is kinetically much less feasible, i.e., the direct mechanism has barriers of 14.3 and 8.7 kcal mol(-1), whereas the indirect mechanism has barriers of 12.6 and 10.7 kcal mol(-1), respectively. This result is in sharp contrast to the analogous HCO + O2 reaction, where the direct (with a barrier of 2.98 kcal mol(-1)) and indirect (2.26 kcal mol(-1)) H-transfer processes are highly competitive over the indirect O-transfer process (the least endothermicity is 19.9 kcal mol(-1)). The possible explanations and implications of the present results are provided.

Algorithms↗
PubMedDate not supplied

Active components of intestinal bacteria for abdominal irradiation-induced inhibition of lung metastases.

We have previously reported that abdominal irradiation of mice inhibited lung metastases of a weakly immunogenic fibrosarcoma, and that transmigration after the irradiation of Enterobacter cloacae into mesenteric lymph nodes coincided with this phenomenon. In this paper, we show that Escherichia coli as well as E. cloacae reduce the number of metastatic lung colonies when these bacteria were intravenously injected into mice prior to the tumour cell challenge. The inhibition was caused not only by the administration of living bacteria but also by that of killed bacteria. Bacterial lipopolysaccharide (LPS), a component of membrane, replaced at least in part the effect of whole bacteria. Transfer of spleen cells from LPS-treated mice into intact recipients prominently inhibited metastatic development in the recipient mice. 'Cross transfer' between LPS high responders and LPS low responders suggested an indirect activity of transferred spleen cells. The antimetastatic activity of LPS depended on the tumour cell type; metastasis of fibrosarcomas was extensively inhibited by LPS irrespective of tumour immunogenicity while that of adenocarcinomas was only slightly inhibited. These results suggest that non-immunological mechanisms are involved in the antimetastatic activity of LPS.

Abdomen↗
PubMed2000

1999 George Lyman Duff memorial lecture: lipid transfer proteins, HDL metabolism, and atherogenesis.

Plasma high density lipoprotein (HDL) levels show an inverse relationship to atherogenesis, in part reflecting the role of HDL in mediating reverse cholesterol transport. The transfer of HDL cholesterol to the liver involves 3 catabolic pathways: the indirect, cholesteryl ester transfer protein (CETP)-mediated pathway, the selective uptake (scavenger receptor BI) pathway, and a particulate HDL uptake pathway. The functions of the lipid transfer proteins (CETP and phospholipid transfer protein) in HDL metabolism have been elucidated by genetic approaches in humans and mice. Human CETP deficiency is associated with increased HDL levels but appears to increase coronary artery disease risk. Phospholipid transfer protein deficiency, produced by gene knockout in mice, results in decreased HDL levels, reflecting decreased transfer of phospholipids from triglyceride-rich lipoproteins into HDL. Obese (ob/ob) mice have markedly increased HDL levels and represent an interesting model of defective HDL catabolism in the liver. In hepatocytes of wild-type mice, there is extensive uptake and resecretion of HDL and selective uptake of cholesteryl ester from HDL during recycling. In ob/ob mice, these processes are defective, suggesting that HDL recycling plays an important role in holo-HDL catabolism, selective uptake, and the determination of plasma HDL levels.

Animals↗
PubMed1988

Potentiation of leukotriene production following sequestration of neutrophils in isolated lungs: indirect evidence for intercellular leukotriene A4 transfer.

Granulocyte (polymorphonuclear leukocyte, PMN) sequestration in the microvascular bed with release of different mediators has been implicated in the pathogenesis of inflammatory and allergic disorders in many organs including the lung. In the present study, we investigated the profile and quantity of leukotriene (LT) generation in isolated blood-free perfused rabbit lungs, in isolated PMNs in vitro and in rabbit lungs, following administration of PMNs, mimicking pulmonary leukostasis. Following stimulation with increasing concentrations of the calcium ionophore A 23137 (0.1 to 2 mumol/L), LTs were detected in the buffer fluid by their chromatographic mobility in different high-performance liquid chromatography (HPLC) systems, by on-line peak spectrum analysis, and by post-HPLC radioimmunoassay (RIA). In isolated lungs, a dose-dependent generation of cysteinyl LTs greater than LTB4, in the complete absence of omega-oxidation products of LTB4 as well as nonenzymatic hydrolysis products of LTA4, was evoked. PMNs in vitro showed a typical profile of LT liberation (LTB4, 20-OH-, and COOH-LTB4, nonenzymatic LTA4 metabolites). In the model of pulmonary leukostasis, the presence of omega-oxidation products of LTB4 indicated metabolic integrity of the trapped PMNs. Nonenzymatic hydrolysis products of LTA4 were, however, not detected in the combined system, whereas the cysteinyl LTs increased markedly. This profile suggests intercellular transfer of PMN-derived LTA4 to lung cells in the microenvironment. In addition, at 2 mumol/L A23187, the sum of all LTA4-derived products surpassed the arithmetic sum of the isolated preparations more than threefold. This potentiation of an LT generation under conditions of pulmonary leukostasis may be of biologic significance for amplification of inflammatory events.

Animals↗
PubMed1990

Xylem-to-Phloem Transfer of Organic Nitrogen in Young Soybean Plants.

Xylem-to-phloem transfer in young vegetative soybean (Glycine max [L.] Merr.) plants (V4 stage) was identified as the difference in the distribution of [(14)C]inulin, a xylem marker, and [(14)C]aminoisobutyric acid (AIB), a synthetic amino acid, fed via the transpiration stream. Since [(14)C]AIB was retained in the stem to some extent, whereas [(14)C]inulin was not, the distribution of these marker compounds in each leaf was expressed as a percentage of the total [(14)C] radioactivity recovered in the foliage. The developing third trifoliolate was a consistent and reliable indicator of xylem-to-phloem transfer. The phloem stream provided to the developing trifoliolate up to fourfold the relative proportion of solute received from the xylem stream; this was markedly reduced by increased light intensity and consequently water flow through the xylem. Evidence from heat girdling experiments is discussed with respect to the vascular anatomy of the soybean plant, and interpreted to suggest that direct xylem-to-phloem transfer in the stem, in the region of the second node, accounted for about one-half of the AIB supplied to the developing trifoliolate, with the remainder being provided from the second trifoliolate. Since AIB is not metabolized it seems likely that rapid transfer within the second trifoliolate occurred as direct veinal transfer rather than indirect cycling through the mesophyll. This study confirmed that xylem-to-phloem transfer plays a major role in the partitioning of nitrogen for early leaf development.

Journal Article↗
PubMed2005

Characterizing residue transfer efficiencies using a fluorescent imaging technique.

To reduce the uncertainty associated with current estimates of children's exposure to pesticides by dermal contact and indirect ingestion, residue transfer data are required. Prior to conducting exhaustive studies, a screening study to identify the important parameters for characterizing these transfers was designed. A fluorescence imaging system was developed (Ivancic et al., in press) to facilitate collection of surface residue transfer data for repeated contacts. Next, parameters that affect residue transfer from surface-to-skin, skin-to-other objects, and skin-to-mouth were evaluated using the imaging system and the fluorescent tracer riboflavin as a surrogate for pesticide residues. Riboflavin was applied as a residue to surfaces of interest. Controlled transfer experiments were conducted by varying contact parameters with each trial. The mass of a tracer transferred was measured and the contact surface area estimated using video imaging techniques. Parameters evaluated included: surface type, surface loading, contact motion, pressure, duration, and skin condition. Transfers both onto, and off of, the hand were measured. To efficiently identify parameter changes resulting in significant effects, the Youden ruggedness test was used to select the combination of parameters varied in each contact trial. In this way, more than one parameter could be varied at a time and the number of trials required was minimized. Results of this study showed that surface loading and skin condition (significant at alpha=0.05) are among the important parameters for characterizing residue transfers of riboflavin. Duration of contact within the time range investigated does not have a significant effect on transfer of this tracer. Results of this study demonstrate the potential for collecting dermal transfer data using the Ivancic et al. fluorescence imaging system and provide preliminary data to reduce uncertainty associated with estimating dermal exposures resulting from contact with residue-contaminated surfaces. These data will also aid in determining what additional residue transfer data should be collected and what type of microactivity data are needed to estimate dermal and indirect ingestion exposure to residues on household surfaces.

Administration, Cutaneous↗
PubMed2002

Orientational effect on the photophysical properties of quaterthiophene-C60 dyads.

Two quaterthiophene-[60]fullerene dyads in which C60 is singly (4TsC) or doubly (4TdC) connected to the inner beta-position of the terminal thiophene rings have been synthesized. The electronic properties of these donor-acceptor compounds were analyzed by UV/Vis spectroscopy and cyclic voltammetry, and their photophysical properties in solution and in the solid state by (time-resolved) photoluminescence (PL) and photoinduced absorption (PIA) spectroscopy. Both the flexible and geometrically constrained 4TsC and 4TdC dyads exhibit photoinduced charge transfer from the quaterthiophene to the fullerene in toluene and o-dichlorobenzene (ODCB). In toluene, charge transfer occurs in both dyads by an indirect mechanism, the first step of which is a singlet-energy transfer from the 4T(S1) state to the C60(S1) state. In the more polar ODCB, direct electron transfer from 4T(S1) competes with energy transfer, and both direct and indirect charge transfers are observed. The geometrical fixation of the donor and acceptor chromophores in 4TdC results in rate constants for energy and electron transfer that are more than an order of magnitude larger than those of the flexible 4TsC system. For both dyads, charge recombination is extremely fast, as inferred from picosecond-resolved temporal evolution of the excited state absorption of the 4T.+ radical cation both in toluene and ODCB.

Journal Article↗
PubMed2005

Myocardial gene transfer and long-term expression following intracoronary delivery of adeno-associated virus.

Adeno-associated viral vectors (AAV) can direct long-term gene expression in post-mitotic cells. Previous studies have established that long-term cardiac gene transfer results from intramuscular injection into the heart. Cardiac gene transfer after direct intracoronary delivery of AAV in vivo, however, has been minimal in degree, and indirect intracoronary delivery, an approach used in an increasing number of studies, appears to be receiving more attention. To determine the utility of indirect intracoronary gene transfer of AAV, we used aortic and pulmonary artery cross clamping followed by proximal aortic injection of AAV encoding enhanced green fluorescent protein (AAV.EGFP) at 10(11) DNase resistant particles (drp; high-performance liquid chromatography (HPLC)-purified) per rat. Gene expression was quantified by fluorescent microscopy at four time points up to 1 year after vector delivery, revealing 20-32% transmural gene expression in the left ventricle at each time point. Histological analysis revealed little or no inflammatory response and levels of transgene expression were low in liver and undetectable in lung. In subsequent studies in pigs, direct intracoronary delivery into the left circumflex coronary artery of AAV.EGFP (2.64-5.28 x 10(13) drp; HPLC-purified) resulted in gene expression in 3 of 4 pigs 8 weeks following injection with no inflammatory response in the heart. PCR analysis confirmed AAV vector presence in the left circumflex perfusion bed. These data indicate that intracoronary delivery of AAV vector is associated with transgene expression in the heart, providing a means to obtain long-term expression of therapeutic genes.

Animals↗
PubMed2006

Photophysical processes involved within the bichromophoric system 9-benzotriazole-1-ylmethyl-9H-carbazole and its role as an artificial photosynthetic device.

From both steady state and fluorescence lifetime measurements it reveals that due to photoexcitation of benzotriazole (BZ) part of the bichromophore, 9(1-H-benzotriazole-lylmethyl)-9H-carbazole (BHC), singlet-singlet energy transfer takes place to populate the lowest excited singlet of carbazole (CZ). CZ, thus being excited indirectly via energy transfer process, undergoes strong charge transfer (CT) reaction with the surrounding polar medium acetonitrile (ACN). On the other hand, very weak CT band was apparent when CZ part, within BHC, was directly excited. In less polar tetrahydrofuran (THF) and polar benzonitrile (BN) environment, lack of formation of CT band strongly suggests in favor of the electron-accepting behavior of ACN. Moreover, by measuring the emission spectra of BHC in microcrystals and of 30 bilayers mixed LB film at high mole fraction of BHC molecules, the possibility of excimer formation or aggregation has been ruled out. Thus, BHC, when dissolved in ACN, acts as a triad system of BZ-CZ-ACN where BZ acts as an antenna molecule and CZ as a reaction center. The possible role of the bichromophoric system BHC as an artificial photosynthetic or solar energy conversion device has been hinted.

Acetonitriles↗
PubMed1992

Penicillin and cephalosporin biosynthetic genes: structure, organization, regulation, and evolution.

Penicillins and cephalosporins are produced by a wide variety of microorganisms, including some filamentous fungi, many gram-positive streptomycetes, and a few gram-negative unicellular bacteria. All produce these beta-lactam antibiotics by essentially the same biosynthetic pathway. Recently, most of the penicillin and cephalosporin biosynthetic genes have been cloned, sequenced, and expressed. The biosynthetic genes code for enzymes that possess multifunctional peptide synthetase, cyclase, epimerase, expandase, hydroxylase, lysine aminotransferase, and acetyltransferase activities and are organized in chromosomal gene clusters and coordinately expressed. DNA hybridization screens of streptomycetes demonstrate that beta-lactam biosynthetic genes may be more widespread in nature than is indicated by conventional antibiotic screens. They offer the possibility of expanding the search for organisms with potential to make new beta-lactam antibiotics. Attempts to improve current yields of beta-lactams in production strains by introducing into them additional copies of biosynthetic genes have been partially successful. Comparative sequence analysis of bacterial and fungal beta-lactam biosynthetic genes show they share very high sequence identity. A model that explains the similarity of biosynthetic genes from an evolutionary standpoint assumes horizontal gene-transfer between the two groups of organisms. Indirect evidence suggests the transfer occurred from the bacteria to the fungi.

Amino Acid Sequence↗
PubMed1993

Transferable resistance to imipenem in hospital isolates of Pseudomonas aeruginosa.

We monitored systematically, for more than five years, the eventual transferability of resistance to imipenem in strains of Pseudomonas aeruginosa isolated from patients in Frankfurt University Clinics. Quite recently, four strains have been found which transfer resistance to imipenem to recipient strains of P. aeruginosa. Although in three strains imipenem was the only antibiotic where resistance was transferred directly, the indirect selection analysis showed that, in each instance, determinants of resistance to carbenicillin and kanamycin were co-transferred. The situation in the fourth strain was more complicated. It was resistant to at least ten antipseudomonad antibiotics, and transferred directly not only determinants of resistance to imipenem, but also to carbenicillin and kanamycin, as did the other strains, plus determinants of resistance to ceftazidime and cefotaxime. The origin and mode of spread of resistance determinants in studied strains is briefly discussed.

Conjugation, Genetic↗
PubMed1981

Mammalian cell survival and radiation quality: analysis with allowance for delta tracks.

The RBE for mammalian cell survival is analysed in relation to the distribution of LET. Complex target theory is described, together with a procedure for applying LET distributions to experimental findings in order to estimate the modes of cellular inactivation. Track segment theory is shown to have previously unrecognised limitations. The stepwise method is adopted to calculate the LET distributions for the radiations (alpha-particles, deuterons and x-rays) used by Barendsen et al (1966) in their study of the survival of T1 cells of human origin. Satisfactory fits of theory to experiment are obtained. Three distinctive modes of cellular inactivation, with independent energy transfers (direct and indirect) to complex targets comprising 2, 4 adn 6 elements respectively, appear to be involved overall. At 'low dose', no contribution from the 2-transfer-mode can be discerned; only this mode is reparable. Saturation cross-sections (at high LET) for the 2- and 4-transfer-, but not the 6-transfer-mode, are oxygen-dependent. All three modes of inactivation might entail unrepaired double strand breaks and/or some form of mutation in DNA, at 1, 2 and 3 vulnerable sections, respectively.

Alpha Particles↗
PubMed2004

Cell surface detection of membrane protein interaction with homogeneous time-resolved fluorescence resonance energy transfer technology.

Direct or indirect interactions between membrane proteins at the cell surface play a central role in numerous cell processes, including possible synergistic effects between different types of receptors. Here we describe a method and tools to analyze membrane protein-protein interaction at the surface of living cells. This technology is based on the use of specific antibodies directed against each partner and labeled either with europium cryptate or with Alexa Fluor 647. This allows the measurement of a fluorescence resonance energy transfer (FRET) signal in a time-resolved manner if both antibodies are in close proximity. This approach is here validated using the heterodimeric gamma-aminobutyrate B receptor as a model. We show that after washing out the unbound antibodies, the time-resolved FRET signal can be measured together with the expression level of both partners via the quantification of the donor and the acceptor fluorophores bound to the cells. Thanks to the high sensitivity of this method and to the low concentration of antibodies required, we show that the signal can also be measured directly after the incubation period without washing out the unbound antibody (homogeneous time-resolved FRET). As such, this method is highly sensitive, reproducible, and compatible with the development of high-throughput screening protocols.

Antibodies↗
PubMed2003

Lateral gene transfer (LGT) between Archaea and Escherichia coli is a contributor to the emergence of novel infectious disease.

BACKGROUND: Lateral gene transfer is the major mechanism for acquisition of new virulence genes in pathogens. Recent whole genome analyses have suggested massive gene transfer between widely divergent organisms. PRESENTATION OF THE HYPOTHESIS: Archeal-like genes acting as virulence genes are present in several pathogens and genomes contain a number of archaeal-like genes of unknown function. Archaea, by virtue of their very different evolutionary history and different environment, provide a pool of potential virulence genes to bacterial pathogens. TESTING THE HYPOTHESIS: We can test this hypothesis by 1)identifying genes likely to have been transferred (directly or indirectly) to E. coli O157:H7 from archaea; 2)investigating the distribution of similar genes in pathogens and non-pathogens and 3)performing rigorous phylogenetic analyses on putative transfers. IMPLICATIONS OF THE HYPOTHESIS: Although this hypothesis focuses on archaea and E. coli, it will serve as a model having broad applicability to a number of pathogenic systems. Since no archaea are known vertebrate pathogens, archaeal-like transferred genes that are associated with virulence in bacteria represent a clear model for the emergence of virulence genes.

Archaea↗
PubMed1991

Comparison of impression techniques for a five-implant mandibular model.

Production of an accurate dental cast framework that meets the implant objective of passive fit demands an understanding of potential processing errors. Working casts fabricated from impressions using two different transfer copings, as provided by a leading implant manufacturer, were investigated for differences in accuracy. A five-implant mandibular model was used to produce seven casts by both the indirect and direct transfer coping techniques. Comparison was made by using a dental cast framework fitted to the master cast. Differences in distances measured between each group and the master cast were analyzed to establish differences between methods. For the model used, the direct technique produced more accurate working casts. A concern for potential impression distortion given the specific clinical situation underscores the method of choice.

Analysis of Variance↗