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PubMedDate not supplied

[Echography applied in dermatologic diseases. A comparative preliminary study of the use of high-frequency transducers].

Ultrasonography (US) was first used in dermatology in 1979. During the last few years, US has evolved thanks to technological progress and its use has been on the increase, in dermatology, to study both localized and diffuse skin diseases. In this paper, the authors review and compare the results obtained with a 20 MHz transducer with those obtained with 7.5 MHz and 10 MHz transducers in the study of skin alterations. Three parameters were considered: lesion identification, spatial evaluation and US structure. Fifty-eight cases of localized and 48 of diffuse skin diseases were examined. The localized forms were benign in nature in 18 patients (3 nevi, 2 seborrheic keratoses, 3 ulcus cruris cases, 6 sebaceous and 2 liquid cysts). The other 40 patients had malignant neoplastic lesions (16 epitheliomas, 13 primitive melanomas, 1 metastatic melanoma, 8 lymphomas and 2 Kaposi's sarcomas). The 48 cases of diffuse conditions included 27 systemic sclerosis, 18 psoriasis and 3 lichen planus cases. The comparative analysis of the results obtained using the different types of transducers in the study of localized dermatoses showed no differences in lesion identification and in the depiction of their US features. On the other hand, the 20 MHz transducer was much more accurate than the others as regards the spatial evaluation of the lesions measurable in millimeters, whereas it poorly demonstrated the lesions > 1.5 cm. However, only the 20 MHz transducer could demonstrate the US features of diffuse conditions.

Evaluation Studies as Topic↗
PubMed1996

In-situ calibration of the implantable force transducer.

Recently, an implantable force transducer (IFT) has been introduced [Xu et al. (1992, J. Biomech. Engng 114, 170-177)] which can be used in tight spaces where force recordings with established transducers, such as the buckle-type transducers, are not possible because of impingement artifacts. The IFT is easily implanted in chronic animal preparations; however, calibration of the IFT in terminal experiments has produced unreliable results. The problems of IFT calibration are that minute movements of the transducer within the tendon, slight misalignments of the tendon, or slight errors in the line of pull cause dramatic changes in the IFT voltage output for a given applied calibration load. Here, we propose a method that eliminates the above calibration problems primarily because the target tendon is left in situ, the calibration loads are applied by the muscles which insert into the target tendon, and the transducer is implanted into the target tendon about two weeks prior to calibration. The theoretical and experimental approaches are demonstrated for the cat patellar tendon, but in principle can be performed with any tendon. The results are repeatable, lie within expected values, and reproduce some of the basic properties which have been observed in prior IFT testing.

Algorithms↗
PubMed1996

Rat primary T cells expressing HTLV-I tax gene transduced by a retroviral vector: in vitro and in vivo characterization.

We prepared a recombinant retroviral vector expressing the human T-lymphotropic virus type-I tax gene. Infection of WKA/H rat splenocytes yielded T-cell lines which proliferated continuously in media supplemented with exogenous interleukin-2 (IL-2) after the control cells ceased to grow. The phenotype of these cells closely resembled that of typical adult T-cell leukemia cells and tax-immortalized human T cells; i.e., positive for CD3, CD4 and IL-2 receptor alpha-chain. Chromosomal analysis revealed that about 10% of the tax-transduced T cells had several chromosomal abnormalities. We also performed in vivo characterization of tax-transduced splenocytes by injecting them into newborn syngeneic rats soon after in vitro infection. Maintenance of the injected tax-transduced cell population and in vivo expression of the tax gene was confirmed in the splenocytes of the injected rats by polymerase chain reaction. However, development of obvious disease was not observed in these rats for up to 18 months after inoculation. These results indicate that tax is capable of immortalizing rat mature CD4+ T cells in vitro but may be insufficient for full transformation of these cells in vivo. Our in vivo system using retrovirally tax-transduced rat T cells could facilitate investigation of the additional genetic events that cooperatively transform T cells transduced with tax gene.

Animals↗
PubMed1997

Optical isometric force transducer for measurement of rat skeletal muscle contraction in the NMR spectrometer.

In order to measure force of contraction of the rat gastrocnemius muscle in the NMR spectrometer with a simultaneous observation of 31P NMR spectra, an optical force transducer was constructed using gratings and optical fibers. Muscle contraction was accurately recorded in the spectrometer without interfering with NMR measurements. The half height width of the phosphocreatine (PCr) peak in the presence or absence of the transducer was 5 Hz. Decrease in PCr and increase in inorganic phosphate (Pi) in the muscle were successfully followed during monitoring of the force developed during exercise. The change in the PCr:(PCr + Pi) ratio correlated well with the time-force integral from the optical force transducer. In general, there was no interference with the 31P NMR spectrum when the transducer was attached to rat leg, so this optical force transducer should prove valuable for studying muscle physiology in the NMR spectrometer.

Animals↗
PubMed1993

The methyl-accepting transducer protein HtrI is functionally associated with the photoreceptor sensory rhodopsin I in the archaeon Halobacterium salinarium.

We have investigated the functional relationship between two proteins involved in the photosensory system of the archaeon Halobacterium salinarium: the photoreceptor sensory rhodopsin I (SRI) and the halobacterial transducer rhodopsin I (HtrI), which has been proposed to be the putative signal transducer of SRI, by genomic DNA analysis of two independent SRI negative mutants, Pho81 and D1. Southern and PCR analyses revealed that both strains bear alterations in the 5' flanking region of the gene encoding SRI, sopI. DNA sequence analysis confirmed the occurrence in this region of htrI, the gene encoding the putative transducer protein. PCR and Northern analyses have shown further that sopI and htrI are expressed as a single transcriptional unit, thus explaining the lack of SRI in mutants with a defective htrI. Expression of the cloned sopI under the control of a heterologous promoter did not restore the SRI-dependent photoresponse in the strain Pho81. Moreover, the photocycling rate of the expressed pigment was clearly lower than in wild type. HtrI is therefore essential for SRI function and most likely modulates the photochemical properties of the photoreceptor via direct physical interaction. Finally, reintroduction of both sopI and htrI into Pho81 and D1 restored the SRI photochemistry and its physiological function. Our results provide the first experimental evidence for the functional coupling between SRI and HtrI and corroborate the proposed model in which HtrI acts as the signal transducer of this archaeal seven-helix photoreceptor in a way analogous to the bacterial chemotaxis transducers.

Bacterial Proteins↗
PubMed1993

Transduced endothelial cells expressing high levels of tissue plasminogen activator have an unaltered phenotype in vitro.

Elevated cellular plasminogen activator activity has been associated with significant alterations in the in vitro phenotype of both malignant cell lines and nonmalignant endothelial cells. To examine the role of elevated cellular plasminogen activator activity in the production of altered endothelial cell behavior, bovine coronary artery endothelial cells were transduced with a retroviral vector expressing large amounts of tissue plasminogen activator. Cells transduced with the tissue plasminogen activator vector were compared with both untransduced cells and cells transduced with a control vector in a series of in vitro assays of cellular attachment, proliferation, migration, and invasion. The morphology of the 2 transduced populations was unchanged. There was a small decrease (5-15%) in the horizontal migration rate of both transduced cell populations, as compared with that of untransduced cells. No significant differences were detected among the three cell populations in any of the other assays. We conclude that expression of high levels of tissue plasminogen activator does not specifically affect endothelial cell phenotype in vitro. These data lend support to the hypothesis that elevated plasminogen activator activity is necessary but not sufficient to produce alterations in endothelial cell behavior.

Animals↗
PubMed2003

Inhibition of HIV-1 replication in primary human T cells transduced with an intracellular anti-HIV-1 p17 antibody gene.

BACKGROUND: Previously we reported that human CD4(+) T cell lines stably expressing anti-HIV-1 gag p17 scFv/Ckappa in the cytosol or nucleus were resistant to HIV-1 challenge. Inhibition of HIV-1 by anti-HIV-1 gag p17 scFv/Ckappa occurred at both the pre- and post-integration steps of the viral cycle. To simulate more closely the in vivo infection process, in this study we tested anti-HIV-1 activity of anti-HIV-1 gag p17 scFv/Ckappa in primary human T cells. METHODS: Anti-HIV-1 gag p17 scFv/Ckappa gene that is targeted into cytoplasm was inserted into a MMLV vector and transfected into packaging cell line PT67. The recombinant virus was used to transduce primary human T cells and human CD4(+) T cell line Jurkat. Following transduction, transduction efficiency, transgene expression, and cell phenotypes were studied. Transduced cells were then challenged with 100 TCID(50) of HIV-1 IIIB and primary isolate 5AO12. Following challenge, HIV-1 replication was monitored by p24 production. RESULTS: Both transduced Jurkat and primary human T cells expressed the transgene. The expression of the transgene did not alter cell growth and CD4 or CD8 expression. However, HIV-1 replication in scFv/Ckappa-transduced Jurkat cells was inhibited by nearly 90% as compared with vector controls. More importantly, HIV-1 replication in primary human T cells from multiple donors transduced with the anti-HIV-1 gag p17 scFv/Ckappa gene was inhibited by as much as 99% as compared with primary T cells transduced with the vector control. The inhibition of replication was not due to interference in viral entry or reverse transcription. The less that HIV-1 replicated in different donor cells, the higher the degree of protection. CONCLUSIONS: The expression of the anti-HIV-1 gag p17 scFv/Ckappa gene construct in primary human T cells renders these cells resistant to HIV-1 and points to the potential clinical usefulness of this gene construct for anti-HIV-1 gene therapy.

Animals↗
PubMed2003

Lentivirally transduced dendritic cells as a tool for cancer immunotherapy.

BACKGROUND: Dendritic cells (DC) are the professional antigen-presenting cells of the immune system, fully equipped to prime naive T cells and thus essential components for cancer immunotherapy. METHODS: We tested the influence of several elements (cPPT, trip, WPRE, SIN) on the transduction efficiency of human DC. Human and murine DC were transduced with tNGFR-encoding lentiviruses to assess the effect of transduction on phenotype and function. Human DC were transduced with lentiviruses encoding huIi80MAGE-A3 and murine DC with huIi80tOVA to test antigen presentation. RESULTS: A self-inactivating (SIN) lentiviral vector containing the trip element was most efficient in transducing human DC. The transduction of DC with trip/SIN tNGFR encoding lentiviral vectors at MOI 15 resulted in stable gene expression in up to 94.6% (murine) and 88.2% (human) of the mature DC, without perturbing viability, phenotype and function. Human huIi80MAGE-A3-transduced DC were able to stimulate MAGE-A3-specific CD4(+) and CD8(+) T cell clones and could prime both MAGE-A3-specific CD4(+) and CD8(+) T cells in vitro. Murine huIi80tOVA-transduced DC were able to present OVA peptides in the context of MHC class I and class II in vitro and induced a strong OVA-specific cytotoxic T lymphocyte response in vivo, that was protective against subsequent challenge with OVA-expressing tumor cells. CONCLUSIONS: We show that, using lentiviral vectors, efficient gene transfer in human and murine DC can be obtained and that these DC can elicit antigen-specific immune responses in vitro and in vivo. The composition of the transfer vector has a major impact on the transduction efficiency.

Animals↗
PubMed2002

Ex vivo and in vitro studies of transgene expression in rat astrocytes transduced with lentiviral vectors.

Implantation of cells genetically modified to express therapeutic genes into the brain has been proposed as a potential treatment for neurodegenerative diseases. In the current study embryonic rat-derived astrocytes were cultured and transduced with a lentiviral vector expressing the reporter gene green fluorescent protein (GFP) and subsequently grafted into the adult rat brain. The proportion of GFP expressing cells was stable, albeit small (1%), at all survival times, up to 6 weeks, the longest time point studied. In parallel in vitro studies, the astrocytes were lentivirally transduced to express either one of the two isoforms of glutamate decarboxylase (GAD(65) or GAD(67)) or glial cell line-derived neurotrophic factor (GDNF). When transducing 293T cells with the two GAD vectors, released GABA could be measured using high-performance liquid chromatography. Further studies of rat astrocytes transduced with the same vectors resulted in a level of GAD activity about 10 times higher than the activity of an intact rat striatum. One hundred thousand astrocytes transduced with LV-GDNF released approximately 27 ng of GDNF per hour. Thus, taken together, our observations provide support for the use of rat astrocytes in ex vivo gene transfer of these proteins in animal models of CNS disorders, e.g., Parkinson's disease or epilepsy.

Animals↗
PubMed1994

Transplantation of lac-Z-transduced microvascular endothelial cells into the skeletal muscle capillary bed of the rat hindlimb occurs independent of the duration of femoral artery occlusion after injection of cells.

The skeletal muscle capillary bed may be an ideal recipient site for transplantation of genetically modified autologous endothelial cells and thus provide a basis for a technique of somatic gene therapy that would be applicable to a variety of acquired and inherited human diseases. The purpose of this study was to test the hypothesis that adhesion of lac-Z-transduced microvascular endothelial cells (MVEC) in the skeletal muscle capillary bed in vivo is dependent on the duration of arterial occlusion after injection of the transduced MVEC. MVEC derived from the abdominal fat pad of syngeneic rats (Wistar F-455) were transfected with the BAG vector, a replication-incompetent retroviral vector containing the lac-Z gene for beta-galactosidase and the Tn5 gene for selection of the transduced cells by the neomycin analogue, G418. lac-Z-transduced MVEC were radiolabeled with 125I-PKH-95, and, after the femoral artery was occluded for 10 min, these cells (1 to 2 x 10(6)) were injected intraarterially into the rat hindlimb. In the experimental groups the femoral artery clamp was removed at 0, 60, or 120 min after injection. A control group without pre- or postinjection femoral arterial occlusion was also studied. Adhesion of MVEC in the skeletal muscle capillary bed (mean percentage of injected 125I activity) was determined in groups of 4 rats at 1 day, 1 week, and 1 month after injection. Adhesion of the transduced MVEC did not increase as the duration of femoral artery occlusion after injection was increased.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed2000

Characterization of anti-CCR5 ribozyme-transduced CD34+ hematopoietic progenitor cells in vitro and in a SCID-hu mouse model in vivo.

The cellular entry of HIV is mediated by the specific interaction of viral envelope glycoproteins with the cell-surface marker CD4 and a chemokine receptor (CCR5 or CXCR4). Individuals with a 32-base-pair (bp) deletion in the CCR5 coding region, which results in a truncated peptide, show resistance to HIV-1 infection. This suggests that the downregulation of CCR5 expression on target cells may prevent HIV infection. Therefore, ribozymes that inhibit the CCR5 expression offer a novel approach for anti-HIV gene therapy. To assess the effect of an anti-CCR5 ribozyme (R5Rbz) on macrophage differentiation, CD34+ hematopoietic progenitor cells were transduced with a retroviral vector carrying RSRbz and allowed to differentiate in the presence of appropriate cytokines. R5Rbz-transduced CD34+ cells differentiated normally into mature macrophages that carried CD14 and CD4 surface markers, expressed the anti-CCR5 ribozyme, and showed significant resistance to viral infection upon challenge with the HIV-1 BaL strain. Using an in vivo thymopoiesis model, the effect of RSRbz on stem cell differentiation into thymocytes was evaluated by reconstituting SCID-hu mice thymic grafts with ribozyme-transduced CD34+ cells. FACS analysis of cell biopsies at 4 and 6 weeks postengraftment for HLA, CD4, and CD8 markers showed comparable levels of reconstitution and similar percentages of subpopulations of thymocytes between grafts receiving R5Rbz-transduced and control CD34+ cells. RT-PCR assays demonstrated the expression of the anti-CCR5 ribozyme in CD4+, CD8+, and CD4+/CD8+ thymocyte subsets derived from RSRbz-transduced CD34+ cells. These results indicate that anti-CCR5 ribozyme can be introduced into hematopoietic stem cells without adverse effects on their subsequent lineage-specific differentiation and maturation. The expression of anti-CCR5 ribozymes in HIV-1 target cells offers a novel gene therapy strategy to control HIV infection.

Animals↗
PubMed2001

BCR-ABL-expressing cells transduced with the HSV-tk gene die by apoptosis upon treatment with ganciclovir.

The potential efficacy of prodrug activation of a transduced suicide gene in a cancer cell may be impaired or enhanced by oncoproteins produced by that cell. In the context of a gene therapy protocol for chronic myeloid leukemia (CML) we examined whether the Bcr-Abl fusion protein would have either of these effects. Thus, the mechanism of cell killing by transfer of herpes simplex virus thymidine kinase (HSV-tk) and subsequent ganciclovir (GCV) treatment was examined in pre-B (TonB210.1) cells and myeloid cells (32D) and in their BCR-ABL-expressing counterparts. HSV-tk-transduced cell lines, either in the presence or in the absence of BCR-ABL expression, became susceptible to GCV at concentrations which were nontoxic to the nontransduced cells. This susceptibility was represented by apoptotic cell death in all cases. Apoptosis was observed after 24 h of treatment with GCV in the tk-transduced parental cells and in the BCR-ABL-expressing TonB210.1 cells but only after a delay of more than 24 h in the 32Dp210 cells compared to 32D. Cell death in the BCR-ABL-expressing clones was preceded by S- and G2/M-phase cell cycle arrest. Activation of FAS/APO-1 and caspase-8 was observed in all the tk-transduced cell lines after GCV treatment. However, the caspase-8 inhibitor Z-IETD-FMK only partially abrogated tk/GCV-induced apoptosis. A possible role for inhibition of Bcl-2 or Bcl-x(L) expression in the apoptosis induced by GCV was observed in the tk-transduced TonB210.1 cells but not in the 32D or 32Dp210 cells. The data demonstrate that expression of the Bcr-Abl oncoprotein does not block the apoptosis induced by the HSV-tk/GCV system, suggesting that this suicide gene therapy strategy could be considered for the treatment of CML in blast crisis.

Animals↗
PubMed1984

The transducer and encoder of frog muscle spindles are essentially nonlinear. Physiological conclusions from a white-noise analysis.

Nonlinear second order white-noise analysis has been applied to the isolated frog muscle spindle. Power (delta 2) of the Gaussian white noise (GWN) and the average prestretch level L were varied and the response of both the isolated receptor potential (transducer) and the action potential (encoder) level were analysed. The standard white-noise method is briefly presented. Particular emphasis, however, is put on the limitations in the range of validity of the method and, consequently, on the use and interpretation of the kernels as a Wiener model. Conclusions in the present paper are within this frame and are mainly of qualitative nature. The analysis reveals that the nonlinear contributions of the model are essential for approximating physiological results, thus ruling out purely linear modelling for this receptor organ. The dependence of the transducer kernels on delta are compatible with the behaviour of a rectifier. Rectification is represented by the lack of hyperpolarization within the isolated receptor potential and is enhanced by the substantial memory in the linear and nonlinear kernels as demonstrated by their extent in time. This is equivalent to low power in high frequencies of the response. Obviously, the hyperpolarizing potentials following each spike counteract the long transducer memory. At the encoder level the memory of the system is strongly reduced. This is achieved by using predominantly high frequency components of the receptor potential for triggering the process of impulse generation, and by the precise coupling and high frequency content of the impulses. This coupling precision is possible because of the sensitivity of the spike-generating mechanism to steep rising transients of the receptor potential and also owing to the reduction in transducer memory by the hyperpolarizing afterpotentials. The preference given to the high frequency components is also read from the structure of the second order transducer kernel and from both the linear and the second order encoder kernels, which allows the most effective input waveform for triggering action potentials to be determined. When the operating point is changed to higher prestretch values, kernel heights increase strongly implying higher response strength of the muscle spindle.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMedDate not supplied

[Recording of ventricular pressure by conventional catheter manometer systems. Efficiency of several combinations of conventional catheters, modern transducers and catheter-flush systems (author's transl)].

The experimentally in vitro determined dynamic response characteristics of 38 catheter manometer systems were uniform in the worst case to 5 c.p.s. and optimally to 26 c.p.s. Accordingly, some systems are only satisfactory for ordinary pressure recording in cardiac rest, while better systems record dp/dt correct up to moderate inotropic stimulation of the heart. In the frequency range of uniform response (amplitude error less +/- 5%) the phase distortion is also negligible. In clinical application the investigator is often restricted to special type of cardiac catheter. In this case a low compliant transducer yields superior results. In all examined systems the combination with MSD 10 transducers is best, whereas the combination with P 23 Db transducers leads to minimal results. An inadequate system for recording ventricular pressure pulses leads in most cases to overestimations of dp/dtmax. The use of low frequency pass filters to attenuate higher frequency artefacts is, under clinical conditions, not suitable for extending the range of uniform frequency response. The dynamic response of 14 catheter manometer systems with two types of continuous self flush units was determined. The use of the P 37 flush unit in combination with small internal diameter catheters leads to serious error in ordinary pressure recording, due to amplitude distortion of the lower harmonics. The frequency response characteristics of the combination of an Intraflow flush system and MSD 10 transducer was similar to the non-flushing P 23 Db transducer feature.

Blood Pressure Determination↗
PubMed2005

Marrow mesenchymal stem cells transduced with TPO/FL genes as support for ex vivo expansion of hematopoietic stem/progenitor cells.

A new marrow-derived mesenchymal stem cell (hMSC) line that could support expansion of hematopoietic stem/progenitor cells (HSPCs) was developed. Primary hMSCs were infected with retrovirus containing Flt-3 ligand and thrombopoietin genes. CD34+ cells from cord blood were expanded with primary hMSCs or transduced hMSCs. The expansion of total nucleated cells, CD34+ cells and mixed colonies containing erythroid and myeloid cells and megakaryocytes for 2 weeks coculture with transduced hMSCs was remarkably increased. The outputs of long-term culture-initiating cells for 2 and 4 weeks coculture with transduced hMSCs were also largely increased. The expansion rates of HSPCs with transduced hMSCs were unchanged for 6 weeks. In contrast, the expansion rates of HSPCs with primary hMSCs declined drastically through 6 weeks. SCID-repopulating cell expansion with transduced hMSCs for 4 weeks was significantly higher than that of uncultured CD34,+ cells and HSPCs expanded with primary hMSCs.

Adult↗
PubMed1988

Doppler ultrasound evaluation of valvar pulmonary stenosis from multiple transducer positions in children requiring pulmonary valvuloplasty.

The modified Bernoulli equation has been used to determine the peak pressure difference across stenotic pulmonary valves. The direction of the poststenotic jet may be eccentric in relation to the axis of the pulmonary artery. Consequently, the maximal velocity obtained from the parasternal transducer position may inaccurately estimate peak pressure difference. Thus, Doppler-derived estimates of pressure difference from the parasternal, subcostal, apical, and suprasternal notch transducer positions were compared with peak-to-peak pulmonary artery to right ventricle catheter withdrawal pressure differences in 24 patients admitted for pulmonary valvuloplasty. Suprasternal, subcostal or apical transducer positions produced higher maximal velocities than the parasternal transducer position in 12 of 24 patients when the studies were performed before cardiac catheterization and in 8 of 12 patients when performed during cardiac catheterization. The Doppler-derived estimates using the highest maximal velocity predicted catheterization pressure difference accurately when the measurements were not performed simultaneously (y = 1.05x - 3.3, r = 0.86, standard error of the estimate +/- 18.7 mm Hg) and the correlation was closer when the measurements were performed simultaneously (y = 1.09x - 2.7, r = 0.97, standard error of the estimate +/- 9.4 mm Hg). The transducer position that yielded the highest maximal velocity in an individual patient was the same before and after valvuloplasty. In all groups, the correlation with pressure at cardiac catheterization was improved by using the highest maximal velocity rather than the parasternal maximal velocity.

Cardiac Catheterization↗
PubMed1975

Hypothesis on the role of liganded states of proteins in energy transducing systems.

In energy transducing systems the direction of energy transfer is proposed to be maintained by the synchronized turnovers of the conformational change of one protein coupling up to affect another. Catalysis by those systems implies, therefore, that under new space restrictions the groups of the transducing enzyme increase and decrease reactivity between themselves, with activatory and/or inhibitory ligands (H+, H2O, metals, etc.) and with the electron shells of the reactant molecules. The exergonic reaction-dependent turnover of the forms of the enzyme within the transition complexes would be maintained, therefore, under asymmetric phase angles of conformational-dependent reactivity that would effectively restrict the microscopic reversibility of transducing systems. Some well known reactions, such as hemoglobins Bohr effect, can be used to illustrate that microscopic (molecular) interactions subject to thermodynamic equilibria laws may similarly paricipate as driving forces in energy transducing sytems. This would allow the thermodynamic description of the role of proton translocation as that of a modificatory force of the structural parameters of proteins. Similarly, the relationship between the liganded states of hemoglobin and its change in conformation has been used to develop an illustrative model relating changes in oxido-reduction of electron carriers to induced-fit effects leading to a sequence of ATPase forms in transition complexes which become stabilized as high energy intermediates under the constraints imposed by the membrane of energy transducing organelles.

Adenine Nucleotides↗
PubMed2006

Release of GABA from sensory neurons transduced with a GAD67-expressing vector occurs by non-vesicular mechanisms.

We have demonstrated that dorsal root ganglion neurons transduced with a recombinant replication-defective herpes simplex virus vector coding for glutamic acid decarboxylase (QHGAD67) release GABA to produce an analgesic effect in rodent models of pain. In this study, we examined the mechanism of transgene-mediated GABA release from dorsal root ganglion neurons in vitro and in vivo. Release of GABA from dorsal root ganglion neurons transduced with QHGAD67 was not increased by membrane depolarization induced by 60 mM extracellular K+ nor reduced by the removal of Ca2+ from the medium. Release of GABA from transduced dorsal root ganglion neurons was, however, blocked in a dose-dependent manner by NO-711, a selective inhibitor of the GABA transporter-1. The amount of GABA released from a spinal cord slice preparation, prepared from animals transduced by subcutaneous inoculation of QHGAD67 in the hind paws, was substantially increased compared to animals transduced with control vector Q0ZHG or normal animals, but the amount of GABA released was not changed by stimulation of the dorsal roots at either low (0.1 mA, 0.5-ms duration) or high (10 mA, 0.5-ms duration) intensity. We conclude that QHGAD67-mediated GABA release from dorsal root ganglion neurons is non-vesicular, independent of electrical depolarization, and that this efflux is mediated through reversal of the GABA transporter.

Analysis of Variance↗