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PubMed1988

Direct and indirect electron transfer between electrodes and redox proteins.

The direct electrochemistry of redox proteins has been achieved at a variety of electrodes, including modified gold, pyrolytic graphite and metal oxides. Careful design of electrode surfaces and electrolyte conditions are required for the attainment of rapid and reversible protein-electrode interaction. The electron transfer reactions of more complex systems, such as redox enzymes, are now being examined. The 'well-behaved' electrochemistry of redox proteins can be usefully exploited by coupling the electrode reaction to enzymes for which the redox proteins act as cofactors. In systems where direct electron transfer is very slow, small electron carriers, or mediators, may be employed to enhance the rate of electron exchange with the electrode. The organometallic compound ferrocene and its derivatives have proved particularly effective in this role. A new generation of electrochemical biosensors employs ferrocene derivatives as mediators.

Electrodes↗

PubMed1985

Kinetic studies of thymidine phosphorylase from mouse liver.

Initial velocity and product inhibition studies of thymidine phosphorylase from mouse liver revealed that the basic reaction mechanism of this enzyme is a rapid equilibrium random bi-bi mechanism with an enzyme-phosphate-thymine dead-end complex. Thymine displayed both substrate inhibition and nonlinear product inhibition, i.e., slope and intercept replots vs. 1/[thymine] were nonlinear, indicating that there is more than one binding site on the enzyme for thymine and that when thymine is bound to one of these sites, the enzyme is inhibited. Furthermore, both thymidine and phosphate showed "cooperative effects" in the presence of thymine at concentrations above 60 microM, suggesting that the enzyme may have multiple interacting allosteric and/or catalytic sites. The deoxyribosyl transferase reaction catalyzed by this enzyme is phosphate-dependent, requires nonstoichiometric amounts of phosphate, and can proceed by an "enzyme-bound" 2-deoxyribose 1-phosphate intermediate. These findings are in accord with the rapid equilibrium random bi-bi mechanism and demonstrate that deoxyribosyl transfer by this enzyme involves an indirect-transfer mechanism. These results strongly suggest that phosphorolysis and deoxyribosyl transfer are catalyzed by the same site on thymidine phosphorylase.

Animals↗

PubMed2001

Stopped-flow kinetic studies of flavin reduction in human cytochrome P450 reductase and its component domains.

The reduction by NADPH of the FAD and FMN redox centers in human cytochrome P450 reductase and its component domains has been studied by rapid-mixing, stopped-flow spectroscopy. Reduction of the isolated FAD-domain occurs in three kinetically resolvable steps. The first represents the rapid formation (>500 s(-)(1)) of a charge-transfer species between oxidized FAD and NADPH. This is followed by an isomerization ( approximately 200 s(-)(1)) to a second charge-transfer species, characterized by a more intense absorption in the long-wavelength region. The third step represents hydride transfer from NADPH to FAD and is accompanied by a change in the tryptophan fluorescence of the FAD-domain. Flavin reduction is reversible, and the observed rate of hydride transfer displays a complex dependence on NADPH concentration. Two-electron-reduced FAD-domain is active in electron transfer reactions with the isolated FMN domain through the formation of a weakly associating electron transfer complex. Reduction of the CPR by NADPH occurs without direct spectral evidence for the formation of charge-transfer species, although the presence of such species is inferred indirectly. Transfer of the first hydride ion leads to the accumulation of a blue di-semiquinoid species of the reductase, indicating rapid transfer of one electron to the FMN domain. The di-semiquinoid species decays on transfer of the second hydride ion. A third phase is seen following prolonged incubation with NADPH and is assigned to a series of equilibration reactions between different redox species of the enzyme as the system relaxes to its thermodynamically most stable state. As with the isolated FAD-domain, the first hydride transfer in the reductase shows a complex dependence on NADPH concentration. At high NADPH concentration, the observed rate of hydride transfer is slow (approximately 20 s(-1)), and this attenuated rate is attributed to the reversible formation of an less active complex resulting from the binding of a second molecule of NADPH. The kinetic data are discussed with reference to the potentiometric studies on the enzyme and its component domains presented in the preceding paper in this issue [Munro, A., Noble, M., Robledo, L., Daff, S., and Chapman, S. (2001) Biochemistry 40, 1956-1963].

Animals↗

PubMed1990

Distribution and metabolism of xylem-borne ureido and amino compounds in developing soybean shoots.

Pulse-chase feeding (30-120 minutes) of (14)C-labeled nitrogenous compounds to cut transpiring shoots was used to investigate the early fate of the major xylem-borne solutes in N(2)-fixing soybean (Glycine max) plants at the V(4) growth stage. By comparison with the foliar distribution of [(14)C]inulin (a xylem marker), it was determined that the phloem supply of allantoin, allantoic acid, asparagine, glutamine, aspartate, and arginine, respectively, provided about 20, 10, three, two, five, and 20 times the (14)C delivered to the developing trifoliolate in the xylem stream. Recovery of unmetabolized asparagine, aspartate, and arginine in this indicator trifoliolate, and significant declines in the percentage of (14)C from allantoic acid and allantoin recovered in the first trifoliolate, provided some support for the direct xylem-to-phloem transfer of these compounds, but did not preclude the involvement of indirect transfer. Data on stem retention and foliar distribution, expressed as a function of the relative xylem sap composition, indicated that ureides provide the major sources of nitrogen to all plant parts. There was no consistent distinction in distribution patterns between pairs of similar anionic and neutral compounds. The extent of xylem-to-phloem transfer among the ureido or the amino compounds was inversely related to its prominence in xylem sap.

Journal Article↗

PubMed2002

[The expanded distant skin flap and its application].

OBJECTIVE: To summarize and present the experiences and results of the expanded distant skin flaps used in plastic and reconstructive operations. METHODS: 102 patients who had undergone expanded distant skin flap transfer were reviewed and followed up. RESULTS: Good results were obtained. Especially when there was no sufficient normal skin close to the lesion or defect, the distant skin flaps were employed. The expanded distant skin flap can be directly transferred as a pedicle skin flap or an island skin flap. It can be indirectly transferred with an intermediary carrier or as a free skin flap. CONCLUSION: The expanded distant skin flaps have many advantages, indications, worthy of recommendation.

Humans↗

PubMed2002

Transfers from older parents to their adult children in Taiwan and the Philippines.

This study examines the extent to which older parents provide material resources and services to their children in the Philippines and Taiwan, and the influence of coresidence on reported transfers between parents and their adult children. The data used in the analyses are from two nationally representative household surveys of persons aged 60 and older in the Philippines and Taiwan. Results show that almost half of older parents in the Philippines provide resources to non-coresident children and only 4 percent of older Taiwanese parents currently make such transfers. When transfers with coresident children are included, older parents show much greater involvement in family support: 69 and 14 percent of older parents in the Philippines and Taiwan, respectively, say they make transfers to children. The difference between the two measures diminishes with age, however, as overall levels of giving decline. Results from multivariate models show that while both measures are associated in similar ways with key factors such as age, health, gender, and economic resources, the association between living arrangements and intergenerational transfers varies across measures. These results suggest that older adults' in these countries tend to exhibit a preference for transfers to the children with whom they live, and that coresidence with grandchildren may be an indirect transfer of services to non-coresident adult children.

Journal Article↗

PubMed1996

A ferric reductase activity is found in brush border membrane vesicles isolated from Caco-2 cells.

Brush border membrane vesicles isolated from Caco-2 cells were used to examine whether there is an apical membrane-associated ferric reductase activity in small intestinal enterocytes. A ferric reductase activity which was dependent on NADH or NADPH as reductants was shown. Reduction of Fe(III) was quantified by the formation of a stable Fe(II)/ferrozine complex. The ferric reductase revealed saturation kinetics with a K(m) of 4.12 +/- 0.65 micromol/L and a Vmax of 3.11 +/- 0.043 nmol/(min.mg protein) for NADH. About 25% of the electrons for the NADH-dependent ferric iron reduction were transferred indirectly from the superoxide anion as verified by the superoxide dismutase inhibitable ferric iron reduction rate. However, the main part of Fe(III) reduction occurs directly by catalyzed electron transfer from NADH to ferric iron through (an) enzyme(s) located in the brush border membrane. The ferric reductase activity was inhibited by Pt(II) and especially p-chloromercuribenzoate. Ferricyanide, which is also reduced by the enzyme, is a competitive inhibitor of the Fe(III)/nitrilotriacetate (NTA) complex with a Ki of 43 micromol/L. These results suggest that brush border membranes of enterocytes possess a ferric reductase that reduces ferric to ferrous iron before the iron is transported through the microvillous membrane.

Caco-2 Cells↗

PubMed1998

Indirect glutamate neurotoxicity.

Indirect glutamate toxicity can be demonstrated by exposing dissociated rat hippocampal cultures to the media of the same culture transiently exposed (1 min) to glutamate (0.5 mM). The toxicity was maximum when the media was collected 5 min after the glutamate exposure. While the primary glutamate toxicity was attenuated by ionotropic glutamate receptor antagonists, the transferred, indirect toxicity was unaffected by the same antagonists. The changes in nuclear morphology indicated chromatin condensation and nuclear fragmentation in both primary and transferred toxicity. The stain for DNA damage by TUNEL method also revealed cells staining positive in both primary and transferred glutamate toxicity. These observations demonstrate that glutamate-induced neurotoxicity can be propagated to the uninjured cells by an unknown toxin released into the extracellular space. This neurotoxin induced both apoptosis and necrosis in cultured rat hippocampal cells.

6-Cyano-7-nitroquinoxaline-2,3-dione↗

PubMed1976

Transfer of substrates across the chloroplast envelope.

The chloroplast represents a relatively autonomous metabolic compartment within the plant cell. It is surrounded by an envelope consisting of two membranes of which the inner membrane is the functional barrier. Utilizing the energy of light the chloroplast is able to synthesize dihydroxyacetonephosphate from carbon dioxide and water. To provide the cell with this substrate, inorganic phosphate is required. In the case of phosphate deficiency the product of CO2 fixation may be temporarily stored within the chloroplast as starch. Specific transport processes across the inner envelope membrane permit the transfer of metabolites between the chloroplast and the cytosol. The phosphate translocator facilitates the export of dihydroxyacetone phosphate in exchange for inorganic phosphate. It also catalyzes a shuttle for inorganic phosphate with 3-phosphoglycerate, permitting the indirect transfer of reducing equivalents and of ATP from the chloroplast to the cytosol. The dicarboxylate carrier transporting various dicarboxylates may be suited for the transfer of reducing equivalents from the cytosol into the chloroplast. The ATP translocator, catalyzing a transport of ATP into the chloroplast in exchange for ADP, appears to be important for providing the chloroplast with ATP during the night phase, as required for the mobilization of starch.

Biological Transport↗

PubMed2002

Risk assessment in regulatory policy making for human and veterinary public health.

Risk assessment is the method of systematically identifying and assessing factors that influence the probability and consequences of a negative event occurring. One responsibility of veterinary medicine is to protect animal and human health. Food animal production uses antibiotics to enhance production. Regulators evaluate new production technology to ensure animal safety and safe, edible products and to make public policy decisions by assessing risks/benefits. The U.S. Food and Drug Administration, Center for Veterinary Medicine's (CVM's) first risk assessment addressed the potential human health impact of campylobacter effects associated with the use of fluoroquinolines in food-producing animals. CVM used the Monte Carlo method to estimate risk byprobability distributions that reflect the uncertainty and variability in the data used for the assessment. Enterococci faecium is a species more likely to be resistant to antibiotics of last resort. Effective control of multidrug-resistant enterococci will requirea better understanding of the transfer of E. faeciumfrom animals to humans and the interaction between E. faecium, the hospital environment, and humans; prudent antibiotic use; better contact isolation in hospitals; and better surveillance. CVM will model these factors in a second, more complex risk assessment designed to examine the indirect transfer of resistance from animals to humans. Use of risk assessments allows researchers, the industry, regulatory authorities, and educators to make better policy decisions regarding antimicrobial use in food animals and humans and the development of resistance. Today the question of whether the use of antimicrobials for growth enhancement infood animals should or should not be terminated for the benefit of human health remains unresolved.

Animals↗

PubMed1991

A new method of the description of the information flow in the brain structures.

The paper describes the method of determining direction and frequency content of the brain activity flow. The method was formulated in the framework of the AR model. The transfer function matrix was found for multichannel EEG process. Elements of this matrix, properly normalized, appeared to be good estimators of the propagation direction and spectral properties of the investigated signals. Simulation experiments have shown that the estimator proposed by us unequivocally reveals the direction of the signal flow and is able to distinguish between direct and indirect transfer of information. The method was applied to the signals recorded in the brain structures of the experimental animals and also to the human normal and epileptic EEG. The sensitivity of the method and its usefulness in the neurological and clinical applications was demonstrated.

Animals↗

PubMed2005

Photoelectrochemical behavior of nanostructured TiO(2) thin-film electrodes in contact with aqueous electrolytes containing dissolved pollutants: a model for distinguishing between direct and indirect interfacial hole transfer from photocurrent measurements.

A model for describing the photoelectrochemical behavior of nanostructured TiO(2) thin-film electrodes in contact with aqueous electrolytes containing dissolved pollutant species is presented. The model correlates the steady-state photocurrent, the illumination intensity, and the specific photooxidation mechanism for dissolved pollutant species in competition with water molecules. For physical events, the model considers the existence of hole trapping at surface states and the specific hole-transfer mechanisms taking place at the semiconductor-electrolyte interface (direct, via photogenerated valence-band free holes or indirect, via photogenerated surface-bound OH radicals or both). The model has been applied to the study of the photooxidation of aqueous solutions of formic acid and methanol. In agreement with previous results concerning massive polycrystalline electrodes, methanol is found to be photooxidized via indirect hole transfer, whereas formic acid is photooxidized mainly with the participation of valence-band free holes.

Journal Article↗

PubMed1988

Lipid interactions with in vitro development of mammalian zygotes.

Rabbit zygotes were tested for their ability to sequester radiolabeled acetate, oleate, and arachidonate in intracellular lipid. Radiolabeled arachidonic acid was concentrated 170 +/- 28-fold (mean +/- SEM) and oleic acid was concentrated 105 +/- 26-fold in zygotic lipids during 6 hr of culture when compared with the initial concentrations in culture medium. Acetate was not concentrated into lipids by cultured zygotes. Both long chain fatty acids were incorporated mainly as triglyceride. Polydimethylsiloxane fluid, used to cover the microdroplets of medium during culture, demonstrated lipophilic properties. This characteristic was utilized to indirectly transfer lipids to culture medium, permitting examination only of lipoidal properties of test extracts on embryonal development. For rabbit zygotes, blood plasma extract was detrimental and whole blood extract was beneficial for embryonal cleavage rates during the first 24 hr of culture. A higher proportion of mouse zygotes developed to blastocysts when cultured in modified Ham's F-10 medium compared to BMOC medium, and this difference was negated by inclusion of a lipid extract prepared from rabbit oviductal fluid in the culture system. Comparison of fatty acid analyses of the lipid extracts with development rates of zygotes suggests that modified rates of embryo development may be associated with ratios of individual fatty acids presented to the culture medium rather than with the presence of any single fatty acid.

Acetates↗

PubMed1987

Bilirubin inhibition of enzymes involved in the mitochondrial malate-aspartate shuttle.

The effect of increasing bilirubin concentrations upon the catalytic activity of a series of dehydrogenases and aminotransferases was examined. The particular enzymes were chosen to examine the effect of bilirubin upon the activity of enzymes responsible for the indirect transfer of reducing equivalents across the inner mitochondrial membrane. Malate dehydrogenase was inhibited at very low concentrations of bilirubin and showed competitive inhibition with respect to coenzyme of 2 microM, while the cytosolic form of this enzyme exhibited a 15 microM inhibition constant. Cytosolic glycerol-3-phosphate dehydrogenase was not appreciably inhibited by bilirubin. Both the mitochondrial and cytosolic forms of aspartate aminotransferase showed moderate competitive bilirubin inhibition with respect to substrates with a Ki of 30 microM with respect to 2-oxoglutarate and a Ki of 80 microM with respect to aspartate. Preincubation studies indicated that inhibition was reversible for all enzymes examined. These results are interpreted in terms of the inhibition of the malate-aspartate shuttle by relatively low concentrations of bilirubin.

Animals↗

PubMed1976

The indirect deltopectoral flap.

A method of indirect transfer of the deltopectoral flap is described to repair craniofacial defects out of reach of the conventional direct flap.

Aged↗

PubMed2000

Acoustic energy affects human gingival fibroblast proliferation but leaves protein production unchanged.

BACKGROUND, AIMS: Sonic toothbrushes are well-established in oral home care for plaque removal; however, the effects of low frequency acoustic (sonic) energy released from sonic toothbrushes to the cells of the periodontium have not been investigated. The purpose of this study was to evaluate the effects of sonic energy on human gingival fibroblast proliferation and protein production in cell culture. METHODS: Direct and indirect transfer calibration studies found the fundamental frequency of the Sonicare sonic toothbrush to be 261 hertz (Hz) with amplitudes ranging from 70 to 104 decibels (dB) in the human periodontium. Using an in vitro delivery system, which coupled a signal-wave generator with a bone transducer to mimic the energy delivered by the Sonicare toothbrush, the effects of signal, amplitude and duration were evaluated longitudinally using a gingival fibroblast cell culture model. 8 strains of fibroblasts isolated from healthy human gingiva were seeded at 30,000 cells/35 mm culture dish in minimum essential medium supplemented with 10% fetal bovine serum. To ascertain the relationship of the amplitude and the duration of sonic stimulation to cellular proliferation, gingival fibroblasts were subjected 2x daily to 261 Hz sound at various amplitudes (67-97 dB) for 0, 15, 30, 60, and 120 s on days 1, 3, 5, 7, and 10. RESULTS: It was found that either 30 or 120 s of sound exposure for 10 days of treatment had significant effects on cell proliferation in comparison to control cultures. Specifically, at day 10, 87 dB at 261 Hz for 30 s 2x daily resulted in a 25.5% increase in cell number (p<0.001), whereas 87 dB at 261 Hz for 120 s twice daily caused a 30.9% decrease in cell number (p<0.001) when compared to control cultures. When cells are stimulated under optimum acoustic conditions for 10 days, there was no difference between the treatment and control groups for collagen (p=0.897) or noncollagen (p=0.697) protein production. CONCLUSIONS: Sonic energy has been shown to both increase and decrease cellular proliferation depending on exposure time; however, during optimum sound-induced conditions for cellular proliferation, sonic energy had no effect on fibroblast protein production. These data suggest that sonic energy can affect the behavior of cells in culture. Further research into the mechanisms of these changes will provide important information for manipulating cellular behavior.

Analysis of Variance↗

PubMed2002

Synergistic effects of physical and chemical guidance cues on neurite alignment and outgrowth on biodegradable polymer substrates.

This article demonstrates that directional outgrowth of neurites is promoted by applying a combination of physical and chemical cues to biodegradable polymer substrates. Films of poly-D,L-lactic acid and poly(lactide-co-glycolide) were micropatterned to form grooves on substrate surfaces, using novel indirect transfer techniques developed specifically for biodegradable polymers that cannot be micropatterned directly. Laminin was selectively adsorbed in the grooves. Whole and dissociated dorsal root ganglia were seeded on the substrates and neurite outgrowth and alignment along the microgrooves were measured. The microgrooves provide physical guidance, whereas laminin provides chemical cues to the neurons. The groove depth and spacing were found to significantly influence neurite alignment. The presence of laminin was found to promote neurite adhesion and outgrowth along the grooves. Using a combination of optimized physical and chemical cues, excellent spatial control of directional neurite outgrowth, with up to 95% alignment of neurites, was obtained. The synergistic effect of physical and chemical guidance cues was found to be more effective than individual cues in promoting directional outgrowth of neurites.

Animals↗

PubMed2006

Direct transport within an organized state trauma system reduces mortality in patients with severe traumatic brain injury.

BACKGROUND: Prehospital management of traumatic brain injury (TBI) and trauma system development and organization are aspects of TBI care that have the potential to significantly impact patient outcome. This multi-center study was conducted to explore the effect of prehospital management decisions on early mortality after severe TBI. METHODS: This report is based on 1449 patients with severe TBI (GCS<9) treated at 22 trauma centers enrolled in a New York State quality improvement (QI) program between 2000 and 2004. The prehospital data collected on these patients include time of injury, time of arrival to the trauma center, mode of transport, type of EMS provider, direct or indirect transport, blood pressure and pulse oximetry values, GCS score, pupillary assessment, and airway management procedures. RESULTS: After exclusion criteria were applied, a total of 1,123 patients were eligible for analysis. The majority of patients were male (75%) with a mean age of 36 years. After controlling for arterial hypotension, age, pupillary status, and initial GCS score, direct transport was found to result in significantly lower mortality than indirect transport. Transport mode, time to admission, and prehospital intubation were not found to be related to 2-week mortality. CONCLUSIONS: The present study provides class II evidence that demonstrates a 50% increase in mortality associated with indirect transfer of TBI patients. Patients with severe TBI should be transported directly to a Level I or Level II trauma center with capabilities as delineated in the Guidelines for the Prehospital Management of Traumatic Brain Injury, even if this center may not be the closest hospital.

Adult↗