Transgenic T cells persist in an adoptive transfer model of mouse liver transplantation tolerance.
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Animals↗
Biomedical subjects
K Lee
Publications and source records attributed to K Lee.
Gabapentin inhibits high-threshold calcium channel currents in cultured rat dorsal root ganglion neurones.
1. This study examined the action of gabapentin (gabapentin,1-(aminomethyl) cyclohexane acetic acid (Neurontin) on voltage-gated calcium (Ca(2+)) channel influx recorded in cultured rat dorsal root ganglion (DRG) neurones. 2. Voltage-gated Ca(2+) influx was monitored using both fura-2 based fluorescence Ca(2+) imaging and the whole-cell patch clamp technique. 3. Imaging of intracellular Ca(2+) transients revealed that gabapentin inhibited KCl (30 mM)-evoked voltage-dependent Ca(2+) influx. Both the duration for 50% of the maximum response (W50) and total Ca(2+) influx were significantly reduced by approximately 25-30% in the presence of gabapentin (25 microM). 4. Gabapentin potently inhibited the peak whole-cell Ca(2+) channel current (I(Ba)) in a dose-dependent manner with an estimated IC(50) value of 167 nM. Block was incomplete and saturated at a maximal concentration of 25 microM. 5. Inhibition was significantly decreased in the presence of the neutral amino acid L-isoleucine (25 microM) but unaffected by application of the GABA(B) antagonist, saclofen (200 microM), suggesting a direct action on the alpha(2)delta subunit of the Ca(2+) channel. 6. Gabapentin inhibition was voltage-dependent, producing an approximately 7 mV hyperpolarizing shift in current voltage properties and reducing a non-inactivating component of whole-cell current activated at relatively depolarized potentials. 7. The use of specific Ca(2+) channel antagonists revealed a mixed pharmacology of the gabapentin-sensitive current (N-, L- and P/Q-type), which is dominated by N-type current. 8. The present study is the first to demonstrate that gabapentin directly mediates inhibition of voltage-gated Ca(2+) influx in DRG neurones, providing a potential means for gabapentin to effectively mediate spinal anti-nociception.
Acetates↗
Complete genome sequence and comparative analysis of the metabolically versatile Pseudomonas putida KT2440.
Pseudomonas putida is a metabolically versatile saprophytic soil bacterium that has been certified as a biosafety host for the cloning of foreign genes. The bacterium also has considerable potential for biotechnological applications. Sequence analysis of the 6.18 Mb genome of strain KT2440 reveals diverse transport and metabolic systems. Although there is a high level of genome conservation with the pathogenic Pseudomonad Pseudomonas aeruginosa (85% of the predicted coding regions are shared), key virulence factors including exotoxin A and type III secretion systems are absent. Analysis of the genome gives insight into the non-pathogenic nature of P. putida and points to potential new applications in agriculture, biocatalysis, bioremediation and bioplastic production.
Bacterial Proteins↗
Investigating the psychosocial impact of anti-HIV combination therapies.
Developments in anti-HIV medication have meant that people with HIV/AIDS are now living longer, with some authors arguing that HIV should now be defined as a manageable rather than terminal illness. However uncertainty about the long-term efficacy of such treatments remains. This research aimed to examine the psychosocial impact of the new treatments and to explore whether, and in what ways, they affect psychological wellbeing. Clients were also asked about their use of services and whether they thought services should be changing in response to new pharmacological treatments. A semi-structured interview schedule was developed to elicit the views of six service users. From multiple readings of the qualitative data generated, prominent themes were identified suggesting that participants had ambivalent feelings about taking antiretroviral combination therapy and that being well with HIV raised a number of difficult issues. Four main themes were revealed: (1) disruptions in daily living: antiretrovirals as intrusions to life, (2) the tablets as a visible marker for HIV infection, (3) tempered optimism and increasing horizons, (4) an uncertain and fragile future: life without benefits and services. The results were discussed in terms of limitations of the current study and suggestions for further research.
Activities of Daily Living↗
Conversion from the SPEAK to the ACE strategy in children using the nucleus 24 cochlear implant system: speech perception and speech production outcomes.
OBJECTIVE: The main objective of this study was to assess whether speech perception and speech production in children using the Nucleus 24 cochlear implant system improved with a change in speech processing strategy from the SPEAK to the Advanced Combination Encoder (ACE) strategy. The major difference between the two strategies is that ACE uses a higher stimulation rate (in this study the stimulation rate was 900 Hz per channel) compared with the SPEAK strategy, where the stimulation rate is 250 Hz per channel. Information also was obtained regarding the adjustment period after conversion to the ACE strategy. DESIGN: An ABA experimental design was used where scores were initially obtained using the SPEAK strategy' (in the initial A time interval), and subsequently performance was assessed using the ACE strategy (B time interval) and then again with the SPEAK strategy (second A time interval). The duration of the B interval was 10 wk, and the duration for the second A interval was 4 wk. Seven children aged between 9 and 16 yr who had at least 6 mo experience with the SPEAK strategy participated. Open-set monosyllabic CNC word perception in quiet and Speech Intelligibility Test sentence perception in noise was evaluated at the end of each of the time intervals. Word perception was also monitored at fortnightly intervals during the B time interval. Speech production was assessed at the end of the initial A time interval and at the end of the B time interval. RESULTS: Mean word and phoneme scores for open-set words in quiet for the group of seven children were significantly higher with the ACE strategy as compared with the SPEAK strategy scores obtained in both of the A time intervals. For sentences in noise, mean scores using the ACE strategy as well as the SPEAK strategy at the second A evaluation point were significantly higher than the scores using the SPEAK strategy measured at the first A time interval. This suggests that learning effects may have influenced outcomes. For some subjects, an initial decrease in scores was found during the initial 2-wk period after fitting the ACE strategy; however, scores subsequently were found to be similar to or higher than those when using the initial SPEAK strategy. Analysis of speech production assessments showed an improvement in the medial consonant scores after using the ACE strategy. CONCLUSIONS: This study demonstrated that some children were able to benefit from the additional information provided by the ACE strategy as compared with the SPEAK strategy. However, the differences in overall performance between the two strategies appear to be relatively small.
Acoustic Stimulation↗
Globalisation and public health.
At the dawn of the 21st century, globalisation is a word that has become a part of everyday communication in all corners of the world. It is a concept that for some holds the promise of a new and brighter future, while for others it represents a threat that needs to be confronted and counteracted. In the area of public health, a wide range of claims have been made about the various impacts, both positive and negative, that can be attributed to globalisation. In the ever expanding literature on globalisation and health, it has become apparent that considerable confusion is emerging in both the ways that terminology is applied and concepts are defined. The determinants of health are increasingly multisectoral, and in tackling these challenges it is necessary to take a multidisciplinary approach that includes policy analyses in such areas as trade, environment, defence/security, foreign policy, and international law. In assembling the terms for this glossary, we have attempted to demonstrate the richness of the globalisation and public health debate, and in so doing have selected some of the core terms that require definition. We hope that this glossary will help to clarify this interesting and challenging area, and will also serve as a useful entry point to this new debate in public health.
Global Health↗
Not modern-day body-snatching: the response of the public.
At recent presentations on the history of anatomy in the West of Scotland, our group has been asked whether we would regard the revelations of 1999-2001 about organ retention as a modern form of body-snatching. We have compared newspaper reports of the Glasgow Herald from 1823 to 1832, the decade prior to the Anatomy Act of 1832, and the Herald, Sunday Herald and Evening Times from 1999 to 2001. Clearly body-snatchers appropriated whole corpses while the recent troubles concerned individual organs. Body-snatching was illegal while the crisis over organ retention arose from differing expectations between the medical profession and the public. Both practices caused huge public concern and distress to relatives. There are, however, interesting differences between the two sets of reports. The public had been aware of body-snatching for many years prior to the Anatomy Act, which regulated the supply of cadavers, whereas revelations about organ retention came as a shock. In the organ retention crisis, the parents of the children were more organised in supporting each other and in campaigning for change than were the public in the days of the resurrectionists.
Biomedical Research↗
Single cell expression analysis--pharmacogenomic potential.
A fundamental challenge in biology is to correlate physiology with gene expression in specific cell types. This can only be achieved by understanding gene expression at the level of the single cell because, in many systems, each cell has the capacity to express a unique set of genes. Therefore, each cell can be considered to be functionally distinct. A clearer understanding of gene expression differences at such a discrete level provides an opportunity to develop drugs with more targeted pharmacologies or with decreased side effects.
Animals↗
Treatment of severe fetomaternal alloimmune thrombocytopenia with compatible frozen-thawed platelet concentrates.
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Antigens, Human Platelet↗
Complementary signaling pathways regulate the unfolded protein response and are required for C. elegans development.
The unfolded protein response (UPR) is a transcriptional and translational intracellular signaling pathway activated by the accumulation of unfolded proteins in the lumen of the endoplasmic reticulum (ER). We have used C. elegans as a genetic model system to dissect UPR signaling in a multicellular organism. C. elegans requires ire-1-mediated splicing of xbp-1 mRNA for UPR gene transcription and survival upon ER stress. In addition, ire-1/xbp-1 acts with pek-1, a protein kinase that mediates translation attenuation, in complementary pathways that are essential for worm development and survival. We propose that UPR transcriptional activation by ire-1 as well as translational attenuation by pek-1 maintain ER homeostasis. The results demonstrate that the UPR and ER homeostasis are essential for metazoan development.
Animals↗
The sodium channel beta-subunit SCN3b modulates the kinetics of SCN5a and is expressed heterogeneously in sheep heart.
1. Cardiac sodium channels are composed of a pore-forming alpha-subunit, SCN5a, and one or more auxiliary beta-subunits. The aim of this study was to investigate the role of the recently discovered member of the beta-subunit family, SCN3b, in the heart. 2. Northern blot and Western blot studies show that SCN3b is highly expressed in the ventricles and Purkinje fibres but not in atrial tissue. This is in contrast to the uniform expression of SCN1b throughout the heart. 3. Co-expression of SCN3b with the cardiac-specific alpha-subunit SCN5a in Xenopus oocytes resulted in a threefold increase in the level of functional sodium channel expression, similar to that observed when SCN1b was co-expressed with SCN5a. These results suggest that both SCN1b and SCN3b improve the efficiency with which the mature channel is targeted to the plasma membrane. 4. When measured in cell-attached oocyte macropatches, SCN3b caused a significant depolarising shift in the half-voltage of steady-state inactivation compared to SCN5a alone or SCN5a + SCN1b. The half-voltage of steady-state activation was not significantly different between SCN5a alone and SCN5a + SCN3b or SCN5a + SCN1b. 5. The rates of inactivation for SCN5a co-expressed with either subunit were not significantly different from that for SCN5a alone. However, recovery from inactivation at -90 mV was significantly faster for SCN5a + SCN1b compared to SCN5a + SCN3b, and both were significantly faster than SCN5a alone. 6. Thus, SCN1b and SCN3b have distinctive effects on the kinetics of activation and inactivation, which, in combination with the different patterns of expression of SCN3b and SCN1b, could have important consequences for the integrated electrical activity of the intact heart.
Animals↗
The catalytic Sakurai reaction.
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Catalysis↗
CYP1A1, CYP2E1 and GSTM1 genetic polymorphisms. The effect of single and combined genotypes on lung cancer susceptibility in Chilean people.
CYP1A1, CYP2E1 and GSTM1 polymorphisms were evaluated in Chilean healthy controls and lung cancer patients. In the Chilean healthy group, frequencies of CYP1A1 variant alleles for MspI (m2 or CYP1A1*2A) and ile/val (val or CYP1A1*2B) polymorphisms were 0.25 and 0.33, respectively. Frequencies of variant alleles C (CYP2E1*6) and c2 (CYP2E1*5B) for CYP2E1 were 0.21 and 0.16, respectively and frequency for GSTM1(-) was 0.24. The presence of variant alleles for GSTM1, MspI and Ile/val polymorphisms was more frequent in cases than in controls. However, frequencies for the c2 and C alleles were not significantly different in controls and in cases. The estimated relative risk for lung cancer associated to a single mutated allele in CYP1A1, CYP2E1 or GSTM1 was 2.41 for m2, 1.69 for val, 1.16 for C, 0.71 for c2 and 2.46 for GSTM1(-). The estimated relative risk was higher for individuals carrying combined CYP1A1 and GSTM1 mutated alleles (m2/val, OR=6.28; m2/GSTM1(-), OR=3.56) and lower in individuals carrying CYP1A1 and CYP2E1 mutated alleles (m2/C, OR=1.39; m2/c2, OR=2.00; val/C, OR=1.45; val/c2, OR=0.48; not significant). The OR values considering smoking were 4.37 for m2, 4.05 for val, 3.47 for GSTM1(-), 7.38 for m2/val and 3.68 for m2/GSTM1(-), higher values than those observed without any stratification by smoking. Taken together, these findings suggest that Chilean people carrying single or combined GSTM1 and CYP1A1 polymorphisms could be more susceptible to lung cancer induced by environmental pollutants such as polycyclic aromatic hydrocarbons.
Chi-Square Distribution↗
Effects of cadmium and vanadium ions on antigen-induced signaling in CD4(+) T cells.
Heavy metal environmental pollutants modulate antigen-directed responses by T lymphocytes, but the molecular mechanisms by which certain metal ions exert their effects are only poorly understood. We tested the hypothesis that cadmium and vanadium ions alter antigen-induced T cell signal transduction pathways in CD4(+) T helper cells. We used CD4(+) primary T lymphocytes and splenic T cells from DO.11.10 T cell receptor transgenic mice. We determined the effects of cadmium chloride and sodium orthovanadate at concentrations that did not induce apoptotic cell death, but affected cytokine or proliferation responses to antigenic stimulation. We used electrophoretic mobility shift assays to measure effects of cadmium and vanadium ions on antigen-induced activation of the nuclear transcriptional regulator proteins, nuclear factor-kappaB, cyclic AMP response element binding protein, nuclear factor of activated T cells, and activator protein-1. Different signaling pathways lead to activation of these transcription factors. Our results suggest that the two heavy metal ions differentially affect signaling pathways. This knowledge will help in the development of molecular epidemiological assays.
Animals↗
Effects of heavy metal ions on resting and antigen-activated CD4(+) T cells.
Heavy metal environmental pollutants increase susceptibility of affected individuals to bacterial and viral infections, but the mechanisms responsible for this effect are not known. We established cellular in vitro systems to identify molecular targets for the action of heavy metal ions. We used two model systems to determine the effects of heavy metal ions on antigen-induced T lymphocyte responses. The first system was representative of primary antigen responses and utilized CD4(+) primary T lymphocytes derived from DO.11.10 T cell receptor transgenic mice. The second system represented a memory T cell phenotype and utilized the CD4(+) T helper 1 clone, pGL2. We measured the effects of the four heavy metals cadmium, lead, mercury, and vanadium on cytokine and proliferation responses by purified CD4(+) T cell to antigenic stimulation. Cytokine responses were differentially affected by lead and vanadium at concentrations that did not affect T cell proliferation in response to antigen. We also determined whether the metal ions induced apoptotic cell death. Mercury induced apoptosis at concentrations as low as 0.5 microM, whereas cadmium required a concentration of 100 microM. Lead (maximal concentration tested was 200 microM) and vanadium (100 microM) did not induce apoptosis. The results suggested that the different heavy metal ions differentially affected antigen-stimulated responses in T helper cells. These in vitro systems can now be applied to test whether heavy metal ions alter antigen-induced T cell signal transduction pathways in CD4(+) T helper cells.
Air Pollutants↗
Streptozocin-induced neuropathy is associated with altered expression of voltage-gated calcium channel subunit mRNAs in rat dorsal root ganglion neurones.
Voltage-gated calcium channels (VGCCs) within sensory neurones are believed to perform an important role in neuropathic pain. In the present study we examine the changes in VGCC mRNA which occur following streptozocin- (STZ) induced diabetic neuropathy using in situ hybridization. STZ caused a significant increase in alpha(2)delta(1), alpha(2)delta(2), and alpha(2)delta(3) mRNA levels in all neuronal cell types. Similarly, mRNA levels of alpha(1F), alpha(1I), and alpha(1S) were increased in all cell types studied whilst alpha(1A) and alpha(1G) mRNAs were specifically upregulated in medium and large diameter neurones. In conclusion, we demonstrate that the induction of diabetic neuropathy is associated with dramatic changes in the expression of VGCCs.
Animals↗
Adenosine exerts multiple effects in dorsal horn neurones of the adult rat spinal cord.
In the present study, we have examined the effects of adenosine and its analogues on the electrophysiological properties of dorsal horn neurones in the rat adult spinal cord. Adenosine and the A1 receptor agonist R-phenylisopropyl adenosine (RPIA) reversibly hyperpolarised these neurones via the generation of an outward current at -60 mV that was inhibited by pre-application of barium or Rp-adenosine 3', 5'-cyclic monophosphothioate triethylamine. In contrast, the A2a receptor agonist 2-[p-(2-carboxyethyl)phenylethylamino]-5'-N-ethylcarboxamidoadenosine (CGS21680) had no effect on the resting membrane properties of these neurones. Stimulation of the dorsal root evoked non-NMDA receptor-mediated excitatory postsynaptic currents (EPSCs) at -60 mV that were completely abolished by 2,3-dihydroxy-6-nitro-7-sulophamoyl-benzo(F)quinoxalone (NBQX). Bath application of adenosine or RPIA reversibly inhibited these EPSCs in a concentration-dependent manner via a presynaptic action. In contrast, CGS21680 increased the amplitude of the EPSC in 20% of neurones tested and decreased the EPSC amplitude in 30% of neurones tested. It is concluded that adenosine exerts multiple effects upon the electrophysiological properties of dorsal horn neurones in the adult spinal cord via interaction with multiple receptors. These findings have important implications in the understanding of adenosine action in preclinical models of pain.
Adenosine↗
Neoceptor concept based on molecular complementarity in GPCRs: a mutant adenosine A(3) receptor with selectively enhanced affinity for amine-modified nucleosides.
Adenosine A(3) receptors are of interest in the treatment of cardiac ischemia, inflammation, and neurodegenerative diseases. In an effort to create a unique receptor mutant that would be activated by tailor-made synthetic ligands, we mutated the human A(3) receptor at the site of a critical His residue in TM7, previously proposed to be involved in ligand recognition through interaction with the ribose moiety. The H272E mutant receptor displayed reduced affinity for most of the uncharged A(3) receptor agonists and antagonists examined. For example, the nonselective agonist 1a was 19-fold less potent at the mutant receptor than at the wild-type receptor. The introduction of an amino group on the ribose moiety of adenosine resulted in either equipotency or enhanced binding affinity at the H272E mutant relative to wild-type A(3) receptors, depending on the position of the amino group. 3'-Amino-3'-deoxyadenosine proved to be 7-fold more potent at the H272E mutant receptor than at the wild-type receptor, while the corresponding 2'- and 5'-amino analogues did not display significantly enhanced affinities. An 3'-amino-N(6)-iodobenzyl analogue showed only a small enhancement at the mutant (K(i) = 320 nM) vs wild-type receptors. The 3'-amino group was intended for a direct electrostatic interaction with the negatively charged ribose-binding region of the mutant receptor, yet molecular modeling did not support this notion. This design approach is an example of engineering the structure of mutant receptors to recognize synthetic ligands for which they are selectively matched on the basis of molecular complementarity between the mutant receptor and the ligand. We have termed such engineered receptors "neoceptors", since the ligand recognition profile of such mutant receptors need not correspond to the profile of the parent, native receptor.
Adenosine↗