Proceedings of the Facial and Aesthetics Conference and Exhibition, Royal College of Physicians, London, 26-27 June.
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Cosmetic Techniques↗
Biomedical subjects
David Ross
Publications and source records attributed to David Ross.
Quinone reductases multitasking in the metabolic world.
The multiple functions of NAD(P)H:quinone oxidoreductase 1 (NQO1, DT-diaphorase) in the cell are reviewed. NQO1 has long been viewed as a chemoprotective enzyme involved in cellular defense against the electrophilic and oxidizing metabolites of xenobiotic quinones. It also participates in reduction of endogenous quinones, such as vitamin E quinone and ubiquinone, generating antioxidant forms of these molecules. NQO1 has recently been shown to interact with superoxide and may be involved in scavenging superoxide within the cell. In addition, the possible role of NQO1 in p53 stabilization and consequently in contributing to p53-dependent stress responses is summarized. Such protein multitasking is a good strategy in terms of cellular economy. NQO1 can also be exploited in the design of NQO1-directed antitumor agents such as the new aziridinylbenzoquinone RH1 and Hsp90 inhibitors such as 17AAG. Polymorphisms in NQO1 which have profound influence on phenotype such as the NQO1*2 polymorphism may influence the chemoprotective actions of NQO1, and should be considered when NQO1-directed antitumor quinones are used for therapy in patients.
Animals↗
High dose buprenorphine in pregnancy.
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Adult↗
Intramural radiofrequency ablation: effects of electrode temperature and length.
The aim of this study was to evaluate intramural temperature-controlled radiofrequency ablation by determining the intramural temperature profile during ablation and by correlating lesion geometry with intramural electrode size and temperature. Intramural ablation might be useful to create deeper lesions for ventricular tachycardia secondary to underlying heart disease. Intramural radiofrequency ablation was performed in 17 greyhounds at thoracotomy, from an epicardial approach, using a 21-gauge needle electrode. Sixty-eight lesions were created in 11 dogs at electrode temperatures of 70 degrees C, 80 degrees C, 90 degrees C, and 100 degrees C for 60 seconds. Intramural thermocouples at 1-, 2-, 3-, 4-, and 5-mm distances were used to identify simultaneous intramural temperature profile. An epicardial approach was used to ensure accurate positioning of the ablating and temperature monitoring needles within the myocardium with fixed interneedle distances. Ninety-nine radiofrequency ablations were performed in six greyhounds using three different intramural electrode lengths (1 mm, 2.5 mm, and 5.5 mm). Lesions were created at 70 degrees C, 80 degrees C, and 90 degrees C for 60 seconds. All lesions were measured after staining with Gomori Trichrome. Lesion dimensions increased in a highly predictable manner with increasing electrode temperature or length. There was no popping or charring, even with target electrode temperature of 100 degrees C. There was significant correlation between intramural temperature 4 mm from the ablating electrode and lesion width (P < 0.001, R2= 0.45) and depth (P = 0.02, R2= 0.08). Feedback control of electrode temperature enables reliable intramural radiofrequency ablation without impedance rise even with target electrode temperature of 100 degrees C. Increasing the length of the intramural ablating electrode to > or = 5.5 mm and increasing temperatures to 90 degrees C-100 degrees C creates the largest lesions.
Animals↗
NAD(P)H:quinone oxidoreductase 1: role as a superoxide scavenger.
Experiments using purified recombinant human NAD(P)H:quinone oxidoreductase 1 (NQO1) revealed that the auto-oxidation of fully reduced protein resulted in a 1:1 stoichiometry of oxygen consumption to NADH oxidation with the production of hydrogen peroxide. The rate of auto-oxidation of fully reduced NQO1 was markedly accelerated in the presence of superoxide (O(2)(*)(-)), whereas the addition of superoxide dismutase greatly inhibited the rate of auto-oxidation. The ability of reduced NQO1 to react with O(2)(*)(-) suggested a role for NQO1 in scavenging O(2)(*)(-), and this hypothesis was tested using established methods for O(2)(*)(-) production and detection. The addition of NQO1 in combination with NAD(P)H resulted in inhibition of dihydroethidium oxidation, pyrogallol auto-oxidation, and elimination of a potassium superoxide-generated ethoxycarbonyl-2-methyl-3,4-dihydro-2H-pyrrole-1-oxide:O(2)(*)(-) adduct signal (electron spin resonance). Kinetic parameters for the reduction of O(2)(*)(-) by NQO1 were estimated using xanthine/xanthine oxidase as the source of O(2)(*)(-) and after NQO1-dependent NADH oxidation at 340 nm. The ability of NQO1 to scavenge O(2)(*)(-) was also examined using cell sonicates prepared from isogenic cell lines containing no NQO1 activity (NQO1(-)) or very high levels of NQO1 activity (NQO1(+)). We demonstrated that addition of NAD(P)H and cell sonicate from NQO1(+) but not NQO1(-) cells resulted in an increased level of O(2)(*)(-) scavenging could be inhibited by 5-methoxy-1,2-dimethyl-3-[(4-nitrophenoxy)methyl]indole-4,7-dione (ES936), a mechanism-based inhibitor of NQO1. NQO1 can generate hydroquinones that are redox active, and the O(2)(*)(-) scavenging activity of NQO1 may allow protection against O(2)(*)(-) at the site of hydroquinone generation. In addition, the O(2)(*)(-) scavenging activity of NQO1 may provide an additional level of protection against O(2)(*)(-) induced toxicity.
Animals↗
Assessment of the cardiac patient for fitness to fly: flying subgroup executive summary.
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Aircraft↗
Differential expression of the antioxidant response element within the hNQO1 promoter in NSCLC versus SCLC.
To determine whether the human (h) NQO1 (NAD(P)H: quinone oxidoreductase 1) gene contains DNA sequences that directly mediate its high and low expression in non-small cell lung carcinoma (NSCLC) and small cell lung carcinoma (SCLC), respectively, a series of deletion constructs spanning up to -4kb of the 5(') flanking region of hNQO1 was used in transient transfection assays. The antioxidant response element (ARE) was found to be critical to the elevated expression of NQO1 in NSCLC. However, the ability of both heterologous and deletion promoter constructs to confer ARE responsiveness demonstrated that SCLC contains the necessary program/menu of transcription factors responsible to drive hNQO1 expression via the ARE. By examining the regulatory region of the hNQO1 gene, we identified a proximal repressor region between -259 and -131. These results provide the first evidence of a proximal repressor region exerting a negative role on the regulation of the hNQO1 promoter in SCLC.
Antioxidants↗
Association between NAD(P)H: quinone oxidoreductase 1 (NQ01) inactivating C609T polymorphism and adenocarcinoma of the upper gastrointestinal tract.
NQO1 is an antioxidant enzyme, important in the detoxification of environmental carcinogens. A single nucleotide polymorphism (C-->T) at position 609 of the NQO1 cDNA has been associated with susceptibility to tumours induced by chemical carcinogens. In our case-control study, we determined the prevalence of the C609T NQO1 polymorphism by PCR-RFLP analysis in Caucasian patients with oesophageal adenocarcinoma (OAC; n=61), cardiac adenocarcinoma (CAC; n=120) or gastric adenocarcinoma (GAC; n=203) vs. a control group that consisted of 252 healthy blood donors. Additionally, NQO1 mRNA expression and NQO1 protein expression were determined by RT-PCR and immunohistochemistry in a subset of cases. The NQO1 C609T genotype distribution was significantly different among controls (C/C, 73.4%; C/T, 25.0%; T/T, 1.6%) as compared to OAC patients (C/C, 49.2%; C/T, 47.5%; T/T, 3.3%; p=0.0004), CAC patients (C/C, 55.8%; C/T, 40.0%; T/T, 4.2%; p=0.0005) and with GAC patients (C/C, 65.5%; C/T; 30.6%, T/T; 3.9%; p=0.0377). The 609T allele overall frequency was 0.141 in controls, 0.270 in OAC patients, 0.241 in CAC patients and 0.192 in GAC patients. Individuals carrying 1 or 2 609T alleles had a 2.85-fold higher risk (95% CI: 1.61-5.07; p=0.0003) for the development of OAC and a 2.18-fold higher risk (95% CI: 1.38-3.44; p=0.0007) for the development of CAC than wild-type gene homozygotes. Immunohistochemical analysis showed NQO1 protein expression in 133 carcinomas, whereas 17 carcinomas were negative. Negativity for NQO1 protein expression correlated strongly with the NQO1 genotype being present in 3.9% of cases with C/C, 13.9% of cases with C/T and 62.5% of cases with T/T genotype (p<0.001). In contrast, NQO1 mRNA expression was detectable irrespective of underlying genotype. In conclusion, determination of the NQO1 genotype may gain importance as a stratification marker in future prevention trials for adenocarcinoma of upper gastrointestinal tract.
Adenocarcinoma↗
The role of non-viraemic transmission on the persistence and dynamics of a tick borne virus--Louping ill in red grouse ( Lagopus lagopus scoticus) and mountain hares ( Lepus timidus).
There exist many tick borne infections that are of either economic or public health interest. Mathematical models have previously been used to describe the dynamics of these infections. However it has recently come to light that there is an alternative mechanism for the transmission of these diseases that has not been considered in a modelling framework. This is transmission through ticks co-feeding on non-viraemic hosts. This paper extends a simple mathematical model to include this alternative transmission mechanism. The model is used to describe the dynamics of Louping ill virus in red grouse (the viraemic host) and hares (the non-viraemic host). However, these results are applicable to many other systems. The model is analysed using joint threshold density curves. It is found that the presence of a non-viraemic host allows the virus to persist more readily than it would in the presence of a host that simply amplified the tick population. More importantly, if the level of non-viraemic transmission is high enough the virus can persist in the absence of the viraemic host. This result has important implications for the control of tick borne diseases.
Algorithms↗
Kinetics of NAD(P)H:quinone oxidoreductase I (NQO1) inhibition by mitomycin C in vitro and in vivo.
The bioreductive activation of the antitumor quinone mitomycin C (MMC) by NAD(P)H: quinone oxidoreductase 1 (NQO1) is complicated by the ability of MMC to also act as a mechanism-based inhibitor of NQO1 in a pH dependent manner. Inhibition of NQO1 by MMC has been studied in purified enzyme preparations and in cultured cells but has not determined in vivo. In the studies presented here, NQO1 activity was measured in mouse tissues following treatment with MMC or the potent mechanism-based human NQO1 inhibitor 5-methoxy-1,2-dimethyl-[(4-nitrophenoxy)methyl]indole-4,7-dione (ES936). NQO1 activity was significantly decreased at 1, 2, and 4 h following MMC (10 or 20 mg/kg) treatment in kidney and lung but was unchanged in brain, heart, liver, and bladder. ES936 (1 mg/kg) treatment led to a significant and much more potent inhibition of NQO1 in all murine tissues analyzed except for bladder. To extrapolate these in vivo results from mice to humans, the species-specific kinetics of NQO1 inactivation by MMC was determined in vitro using mouse, rat, and human recombinant NQO1 proteins. Results showed the inactivation kinetics of mouse and human proteins by MMC were similar. Treatment of human and murine endothelial cells with MMC or ES936 showed similar inhibition of NQO1 activity. The aforementioned results clearly demonstrate that MMC can serve as a substrate for NQO1 in vivo; however, the metabolism resulting in enzyme inactivation is possibly tissue-specific. Furthermore, the kinetic similarities for inactivation between murine and human forms of NQO1 show these results are apropos to clinical use of MMC.
Animals↗
Effect of caged fluorescent dye on the electroosmotic mobility in microchannels.
We report on measurements of electroosmotic mobility in polymer microchannels and silica capillaries with and without the addition of a caged fluorescein dye to the buffer. For PMMA microchannels, the mobility was found to increase from (2.6 +/- 0.1) x 10(-4) cm2 V(-1) s(-1) to (4.6 +/- 0.1) x 10(-4) cm2 V(-1) s(-1) upon addition of 1.2 mmol/L of caged dye. For PC microchannels, the mobility increased from (4.3 +/- 0.2) x 10(-4) cm2 V(-1) s(-1) to (5.4 +/- 0.1) x 10(-4) cm2 V(-1) s(-1) upon addition of caged dye. For PDMS microchannels, the mobility increased from (4.3 +/- 0.2) x 10(-4) cm2 V(-1) s(-1) to (6.4 +/- 0.5) x 10(-4) cm2 V(-1) s(-1) upon addition of caged dye. For fused-silica capillaries, the mobility ((5.5 +/- 0.2) x 10(-4) cm2 V(-1) s(-1)) was unaffected by the addition of the caged dye.
Journal Article↗
Interaction of human NAD(P)H:quinone oxidoreductase 1 (NQO1) with the tumor suppressor protein p53 in cells and cell-free systems.
NAD(P)H:quinone oxidoreductase 1 (NQO1) has been proposed to stabilize p53 via a redox mechanism involving oxidation of NAD(P)H as a consequence of the catalytic activity of NQO1. We report that treatment of HCT-116 human colon carcinoma cells with the NQO1 inhibitor ES936 had no effect on the levels of p53 protein. ES936 is a mechanism-based inhibitor of NQO1 that irreversibly blocks the catalytic function of the enzyme. This suggests that a redox mechanism involving NQO1-mediated NAD(P)H oxidation is not responsible for the stabilization of p53. We also examined the ability of the NQO1 protein to associate with p53 using co-immunoprecipitation experiments. Results from these experiments demonstrated co-immunoprecipitation of NQO1 with p53 and vice versa. The association between p53 and NQO1 was not affected by treatment of HCT-116 cells with ES936, demonstrating that the association was not dependent on the catalytic activity of NQO1. A comparison of isogenic HCT-116 p53+/+ and HCT-116 p53-/- cells demonstrated an interaction of NQO1 and p53 only in the p53+/+ cells. Experiments performed in an in vitro transcription/translation system utilizing rabbit reticulocyte lysates confirmed the interaction of NQO1 and p53. In these experiments a full-length p53 coding region was used to express p53 in the presence of recombinant NQO1 protein. An association of p53 and NQO1 was also observed in primary human keratinocytes and mammary epithelial cells. In studies where mdm-2 co-immunoprecipitated with p53, no association of mdm-2 with NQO1 was observed. These data demonstrate an association between p53 and NQO1 that may represent an alternate mechanism of p53 stabilization by NQO1 in a wide variety of human cell types.
Cell-Free System↗
Modified reperfusion and ischemia-reperfusion injury in human lung transplantation.
OBJECTIVE: Ischemia-reperfusion injury remains a major cause of mortality and morbidity in clinical lung transplantation. Interaction of activated leukocytes with injured graft endothelial cells participates in the development of ischemia-reperfusion injury. We sought to determine if modification of the reperfusate (with depletion of leukocytes and alteration of its composition) would decrease the incidence of ischemia-reperfusion injury in human lung transplantation when compared with whole blood reperfusion in a historical group of patients. METHODS: Between June 1999 and July 2001, 23 adult patients undergoing lung transplantation consented to modified reperfusion. After implantation, a catheter was inserted into the main or individual pulmonary arteries, and modified reperfusate was administered at a pressure less than 20 mm Hg. The modified reperfusate was depleted of leukocytes, supplemented with nitroglycerin, adjusted for pH and calcium level, and enriched with aspartate, glutamate, and dextrose. After 10 minutes of modified reperfusion, the removal of pulmonary artery clamp or weaning of cardiopulmonary bypass was performed per usual protocol. Age- and diagnosis-matched historical patients served as the control group. Ischemia-reperfusion injury was defined as Pao(2)/Fio(2) < 150 with diffuse infiltrate on the radiograph in absence of other causes. RESULTS: There was no difference in donor age or oxygenation indices, recipient age, the number of patients requiring cardiopulmonary bypass, ischemia time, and recipient oxygenation indices between the modified reperfusate group and the control group. However, none of the patients in the modified reperfusate group developed ischemia-reperfusion injury in contrast to 5 patients in the control group (P <.05). The early survival in the modified reperfusate group was 96% versus 81% in the control group (P = NS). CONCLUSION: This study suggests that modification of the reperfusate content decreases the incidence of ischemia-reperfusion injury in human lung transplantation when compared with whole blood reperfusion in a historical group of patients. Modified reperfusate may allow acceptance of marginal lungs and expansion of the donor pool.
Adult↗
Rationales, obstacles and strategies for local outcome monitoring systems in substance abuse treatment settings.
The need for outcome monitoring systems (OMS) in substance abuse treatment is now such a pervasive position that it has achieved 'motherhood' stature. Yet, for a variety of reasons, implementation of systematic, ongoing monitoring continues to lag in most settings. The position taken in this article is that a local approach to monitoring is a viable alternative to large scale monitoring programs when such programs are unavailable. Moreover, local OMS can supplement these more generic programs by providing data that are more in tune with the orientation and program development of a specific setting. This article argues that local OMS can be developed with a reorganization of customary documentation procedures, a modest investment of personnel and specialized technical resources, a shorter-term view to outcome, and a commitment to evidence-based treatment.
Cost-Benefit Analysis↗
Effects of simultaneous insertion of 66 plunge needle electrodes on myocardial activation, function, and structure.
Transmural recordings using plunge needle electrodes are useful in mapping ventricular tachyarrhythmia, but they interfere with activation sequences or damage the myocardium. This study evaluated the effects of insertion of 66 transmural needles on myocardial activation, structure, and function. Epicardial maps were performed at thoracotomy using a 40-electrode plaque in five mongrel dogs. Sixty-six transmural plunge needles were introduced into the anterior aspect of the septum and left ventricle. Transmural maps of unipolar electrograms were recorded every 15 minutes via 124 electrodes over a 2-hour period. Epicardial maps were repeated after the needles were removed. All recordings were performed during sinus rhythm and ventricular pacing at 300- and 200-ms cycle lengths. Gated heart pool studies were performed preoperatively and 2 weeks after thoracotomy. Programmed ventricular stimulation was performed 2 weeks after thoracotomy. In total, 15,996 electrograms were analyzed. Maximum negative dV/dt of each electrogram and the activation time at each electrode did not change significantly over the 2 hours of needle insertion. After removal of the needles, epicardial maps were unchanged compared to before needle insertion. Mean left ventricular ejection fraction 2 weeks after needle insertion was 59% versus 58% before needle insertion (P=0.9). No dogs had inducible ventricular tachycardia. Histology showed contraction bands of 0.8-mm diameter adjacent to the needle tracks but no scarring. Insertion of 66 closely spaced plunge needles did not distort epicardial or transmural maps. Multiple needles did not result in myocardial scarring, left ventricular dysfunction, or predispose to ventricular tachycardia.
Analysis of Variance↗
Association of NAD(P)H: quinone oxidoreductase 1 (NQO1) C609T polymorphism with esophageal squamous cell carcinoma in a German Caucasian and a northern Chinese population.
NAD(P)H: quinone oxidoreductase 1 (NQO1) is an antioxidant enzyme, important in the detoxification of environmental carcinogens. A single base substitution (C --> T) polymorphism at nucleotide 609 (null-allele) of NQO1 gene impairs stability and function of the NQO1 protein. To investigate the association of this NQO1 polymorphism with susceptibility to esophageal squamous cell carcinoma (ESCC), the NQO1 C609T genotypes were determined by PCR-RFLP analysis in 450 patients with ESCC (257 German Caucasians and 193 northern Chinese) and 393 unrelated healthy controls (252 German Caucasians and 141 northern Chinese). Additionally, NQO1 protein expression was determined by immunohistochemistry in a subset of 74 ESCC (50 German, 24 Chinese). A significant difference in NQO1 C609T genotype distribution was observed between Caucasian healthy controls (C/C, 73.4%; C/T, 25.0%; T/T, 1.6%) and Chinese healthy controls (C/C, 34.0%; C/T, 49.7%; T/T, 16.3%) (chi(2) = 68.40, P < 0.001). The NQO1 T/T genotype significantly increased the risk for developing ESCC in both Caucasian subjects (OR = 4.62, 95% CI = 1.54-13.86) and Chinese subjects (OR = 1.81, 95% CI = 1.04-3.15), compared with the combined C/C and C/T genotypes. In Chinese subjects, this increased susceptibility was pronounced in patients with family history of upper gastrointestinal cancers (OR = 2.18, 95% CI = 1.14-4.17). Immunohistochemical analysis showed NQO1 protein expression in 53 carcinomas, whereas 21 carcinomas were negative. Negativity for NQO1 expression correlated strongly with the NQO1 genotype, being present in 8.6% of cases with C/C, 22.2% of cases with C/T and 100% of cases with T/T genotype (chi(2) = 16.60, P < 0.001). In summary, the association of the NQO1 C609T polymorphism with ESCC in genetically distinct populations makes a strong argument for its importance in carcinogenesis of ESCC in the German Caucasian and the northern Chinese population.
Adult↗
EGF and TGF-alpha expression influence the developmental toxicity of TCDD: dose response and AhR phenotype in EGF, TGF-alpha, and EGF + TGF-alpha knockout mice.
The environmental toxicant 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) produces cleft palate (CP) and hydronephrosis (HN) in mice. The etiology of these defects involves hyperproliferation of epithelial cells of the secondary palatal shelf and ureter, respectively. These effects correlate with altered expression of the epidermal growth factor receptor (EGFR), epidermal growth factor (EGF), and transforming growth factor-alpha (TGF-alpha). In this study, the developmental toxicity of TCDD was examined in EGF, TGF-alpha, and double EGF + TGF-alpha knockout (-/-) and wild type (WT) mice. The influence of background genetics in responsiveness to TCDD was examined using liver 7-ethoxyresorufin-O-deethylase (EROD) activity. Animals were dosed by gavage with 0, 0.2, 1, 5, 24, 50, 100, or 150 micro g TCDD/kg (5 ml/kg) body weight on gestation day 12. The mixed genetic background of WT, EGF (-/-), and EGF + TGF-alpha (-/-) made these mice less responsive to TCDD relative to C57BL/6J and TGF-alpha (-/-), which have a C57BL background. These results show that EGF and TGF-alpha are not required for response to TCDD; however, the specific ligand available to bind EGFR affects the responsiveness to TCDD. EGF (-/-) mice are less responsive for CP, but more sensitive to HN. TGF-alpha (-/-) mice were similar to WT in sensitivity for induction of CP and HN. The responses of EGF + TGF-alpha (-/-) mice were like the WT except at higher doses where sensitivity to CP increased, suggesting that the responses may be mediated by alternative ligands for EGFR that are not functional equivalents of EGF or TGF-alpha. In conclusion, the EGFR pathway is mechanistically important in responses of the embryo to TCDD. Specific ligands confer sensitivity or resistance that are target tissue-dependent.
Abnormalities, Drug-Induced↗
Biochemical, cytotoxic, and genotoxic effects of ES936, a mechanism-based inhibitor of NAD(P)H:quinone oxidoreductase 1, in cellular systems.
The specific involvement of NAD(P)H:quinone oxidoreductase 1 (NQO1) in the bioactivation of quinone prodrugs has been shown through the use of the inhibitor of NQO1, dicoumarol. Disadvantages of using dicoumarol to inhibit NQO1 include its lack of specificity and its competitive mechanism of inhibition. The concentration of dicoumarol required for inhibition of NQO1 varies according to the substrate under evaluation, which may lead to either false conclusions of the involvement of NQO1 or the alteration of other cellular processes. We have reported previously on the chemical and biochemical properties of ES936, a mechanism-based inhibitor of NQO1 in cell-free systems. In this study, we investigated the effects of ES936 in cellular systems. ES936 (100 nM) inhibits more than 95% of NQO1 activity within 30 min and is stable in complete media at this concentration for a minimum of 2 h. The duration of inhibition is cell line-specific because a new protein must be generated for resumption of activity. ES936 abrogates the toxicity of streptonigrin, with greater effects seen in cell lines expressing higher levels of NQO1. ES936 does not inhibit other cellular reductases, nor does it alter cellular levels of acid-soluble thiols. Some evidence of DNA strand breaks was observed at the concentrations of ES936 required for the inhibition of NQO1 activity. From our studies, we propose the use of ES936 (100 nM) as a mechanism-based inhibitor of NQO1 in cellular systems and for use as a component of the routine activity assay for NQO1.
Animals↗