Biomedical subjects
G Johnson
Publications and source records attributed to G Johnson.
Cholinesterase-like catalytic antibodies: reaction with substrates and inhibitors.
We have previously described a catalytic monoclonal antibody, raised against acetylcholinesterase (AChE) and capable of hydrolysing acetylthiocholine. Here, we describe two more such antibodies. All three antibodies were raised against the same antigen, human erythrocyte AChE, a commercial product purified using the cholinesterase anionic site inhibitor, tetramethylammonium. IgG was purified on Protein A-Sepharose, and lack of contamination with AChE or butyrylcholinesterase (BChE) was demonstrated on sucrose density gradients and immunoassay of the fractions. The antibodies recognised AchE and were capable of hydrolysing acetylthiocholine and the larger butyrylthiocholine substrate, and were inactivated by phenylmethylsulphonyl fluoride (PMSF), indicating a serine residue in the active site. K(m), K(cat), K(cat)/K(uncat) and K(cat)/K(m) values were obtained for both substrates. The active sites of the antibodies were probed with anti-cholinesterases known to react with the active and anionic sites of acetyl- and BChE, and the peripheral anionic site of AChE. The antibodies were inactivated to varying degrees by the BChE inhibitors iso-OMPA, ethopropazine and tetracaine, indicating a less sterically constrained site than AChE and the lack of an acyl-binding pocket. They were also partially inhibited by the AChE-specific inhibitors, BW284c51 and propidium. No peripheral anionic site, as seen in AChE, was observed, shown by the almost complete lack of reaction with fasciculin. All three antibodies appear to have structures resembling the anionic sites of the cholinesterases, seen by their inhibition by quaternary and tricyclic compounds. Further work is required to determine whether the catalytic activity shown by these antibodies is germline-encoded, or is the result of complexation of the antigen with an inhibitor at a peripheral site.
Clearance of technetium 99m N-NOET in normal, ischemic-reperfused, and membrane-disrupted myocardium.
BACKGROUND: Technetium 99m-labeled bis(N-ethoxy, N-ethyl dithiocarbamato) nitrido technetium(v) (99mTcN-NOET) is a new neutral cardiac perfusion imaging agent that has been shown to have very high uptake and retention in vitro. The purpose of this study was to determine the clearance kinetics of 99mTcN-NOET in control, ischemic-reperfused, and membrane-disrupted myocardium. METHODS AND RESULTS: After a 100 microCi (3.7 x 10(6) Bq) bolus of 99mTcN-NOET was injected, myocardial clearance was monitored for 1 hour by the use of a sodium iodide detector in 30 isolated, Krebs-Henseleit (KH) perfused rat hearts. Seven hearts were used as controls (group 1). In seven ischemic-reperfused hearts, tracer administration and uptake was followed by 30 minutes of no flow and 1 hour of reflow (group 2). In six additional ischemic-reperfused hearts, tracer administration was followed by deprivation of flow for 1 hour followed by 1 hour of reflow (group 3). Six hearts were perfused with a 0.5% Triton X-100 KH perfusate for 1 hour (group 4). Four hearts were perfused with KH for 10 minutes, followed by cyanide for 10 minutes (group 5). This cycle was repeated three times. Activities remaining in each heart at the end of each experiment were quantitated, and activity at peak uptake was calculated. The 99mTcN-NOET myocardial clearance was near linear in the control (0.6 +/- 0.4) and both ischemic-reperfused groups with virtually no fractional clearance (1.2% +/- 0.6% and 2.1% +/- 0.6%, respectively; p = NS). In the Triton X-100 membrane-disrupted hearts, clearance was substantial (94.2% +/- 4.0%; p < 0.0001 compared with the control and ischemic-reperfused groups). Cyanide treatment produced rapid clearance, which was arrested by a return to the standard KH perfusate. Peak uptake as a percentage of injected dose was 74.9% +/- 1.4% for all groups combined. CONCLUSION: Thus 99mTcN-NOET has extremely high myocardial retention after 1 hour in normal myocardium and is not significantly affected by ongoing myocardial ischemia or reperfusion injury in this model. Clearance is increased markedly in extreme conditions of membrane disruption. These data are consistent with the concept that 99mTc-NOET is localized predominantly in or on cell membranes. 99mTcN-NOET is a promising, new myocardial perfusion imaging agent that exhibits a stable myocardial distribution in the setting of acute developing injury.
Planar imaging of 99mTc-labeled (bis(N-ethoxy, N-ethyl dithiocarbamato) nitrido technetium[V]) can detect resting ischemia.
BACKGROUND: (99m)Tc-labeled (bis(N-ethoxy, N-ethyl dithiocarbamato) nitrido technetium(V)) ([99m]TcN-NOET) is a new lipophilic, neutral-charge cardiac perfusion imaging agent that demonstrates apparent redistribution in animal models and humans. The purpose of this study was to determine whether the kinetics of (99m)TcN-NOET are suitable for the detection of resting ischemia. METHODS AND RESULTS: Microspheres were injected at baseline and simultaneously with (99m)TcN-NOET after a 90% reduction in resting flow in the left circumflex coronary artery in six open-chest canine experiments. The relationship of flow and activity early after injection was determined in one experiment by termination at 10 minutes. The flow ratio (left circumflex/left anterior descending coronary artery) after stenosis fell significantly (0.87 +/- 0.04 vs 0.46 +/- 0.04; p < 0.05). The end-tissue (99m)Tc ratio (0.78 +/- 0.05) was significantly higher than the flow ratio at injection (0.46 +/- 0.04; p < 0.05), indicating substantial redistribution. In vivo imaging was conducted during 2 hours in five experiments, followed by ex vivo imaging. Myocardial clearance from 10 minutes onward was biphasic in left anterior descending and monophasic in left circumflex coronary arteries. Myocardial clearance from 10 to 60 minutes was delayed in left circumflex (35.5% +/- 8.1%) versus left anterior descending coronary arteries (49.2% +/- 8.6%; p < 0.05). No significant difference was observed from 60- to 120-minute clearance. Five of five experiments demonstrated initial defects and complete fill-in at 90 to 120 minutes by qualitative assessment. Quantitation of ex vivo images confirmed significant redistribution. CONCLUSIONS: Resting ischemia caused by moderate to severe stenosis can be detected on scans with (99m)TcN-NOET. Redistribution was near complete in this model by 90 to 120 minutes. (99m)TcN-NOET is a promising new agent for the detection of coronary artery disease in viable myocardium and warrants further investigation.
Technetium 99m-HL-91: a potential new marker of myocardial viability assessed by nuclear imaging early after reperfusion.
OBJECTIVE: 99mTc-HL-91 (Prognox) is a potential new hypoxia-avid myocardial imaging agent. The purpose of this study was to determine whether this tracer would demonstrate increased activity in nonviable myocardium in vivo. METHODS AND RESULTS: A 3-hour left circumflex artery (LCx) occlusion was followed by 1 hour of reperfusion, injection of 99mTc-HL-91 (185 MBq), and 2 hours of gamma camera imaging in 6 open-chest canine experiments. Microspheres were injected during baseline, at occlusion, at the time of tracer injection, and at the end of the experiment. After the animals were killed, heart slices were imaged. Blood flow and tracer activity were determined by well counting. Mean infarct size was 19.2% +/- 2.2% (SEM). All six dogs demonstrated no increased 99mTc-HL-91 myocardial activity other than small foci on in vivo and ex vivo gamma camera images. The mean large region of interest (ROI)-determined LCx/LAD (left anterior descending) ratio was 1.10 +/- 0.03 in vivo, and 1.0 +/- 0.02 ex vivo. Mean clearance curves from LCx and LAD ROI were not significantly different, and 2-hour retention was 15.2% +/- 2.1% for the LCx and 18.6% +/- 2.7% for the LAD (p = NS). ROI clearance curves demonstrated biexponential clearance over the first hour and linear clearance over the second hour. Myocardial blood flow (microspheres) versus well-counted tracer uptake curves were linear with near-zero slopes for viable tissue, nonviable tissue, and mosaic tissue. Blood clearance was triexponential with a 2-hour retention of 7.8% +/- 1.1%. CONCLUSIONS: In contrast to viable ischemic tissue, normal and nonviable myocardium demonstrate similar 99mTc-HL-91 uptake and retention kinetics. This agent warrants further clinical studies in situations where there is a need to differentiate ischemic viable from nonviable myocardium.
HL-91-technetium-99m: a new marker of viability in ischemic myocardium.
BACKGROUND: Technetium 99m-HL91 is a new hypoxia imaging agent that demonstrates increased uptake in ischemic, viable myocardium. This study was performed to determine whether HL91 is taken up by nonviable myocardium. METHODS: Twenty-three Krebs-Henseleit buffer-perfused, isolated rat hearts were studied. Tc-99m-HL91 300 microCi was infused over 10 minutes, followed by a 60-minute clearance. Myocardial activity was monitored by use of an NaI crystal. Four groups were studied: control (flow = 12 mL/min, n = 7), low flow (flow = 1 mL/min, n = 6), no flow/reflow (60 minutes no flow/60 minutes reflow before Tc-99m-HL91 infusion, flow = 12 mL/min, n = 5), and cyanide-treated (before Tc-99m-HL91 infusion, flow = 12 mL/min, n = 5). Injury was assessed by creatine kinase, transmission electron microscopy, and triphenyltetrazolium chloride. RESULTS: Control (no injury) and cyanide-treated (severe injury) hearts demonstrated low uptake (6.3+/-0.5 mean+/-SEM and 5.7+/-1.2 microCi, respectively) and low 60-minute retention (13.8%+/-2.2% and 13.7%+/-3.9%, respectively). Low-flow hearts (minimal injury) demonstrated markedly increased uptake (43.5+/-2.8 microCi, P < .01) and increased 60-minute retention (33.2%+/-2.9%, P < .01) compared with control. No-flow/reflow hearts (moderate injury) demonstrated intermediate uptake (8.7+/-0.5 microCi, P < .05 to control), although retention was not significantly different (18.9%+/-3.5%, P = ns). Severely and rapidly injured myocardium demonstrated Tc-99m-HL91 peak uptake and retention indistinguishable from normal. Moderately injured myocardium demonstrated uptake intermediate between severely injured and low-flow-induced ischemic, viable myocardium. CONCLUSION: Thus Tc-99m-HL91 is not taken up or retained in nonviable and irreversibly injured myocardium.
Ocular findings in HIV-1 positive and HIV-1 negative participants in a rural population-based cohort in Uganda.
PURPOSE: To report the ocular findings in HIV-1 seropositive individuals and HIV-negative controls in a population-based cohort in rural Uganda. METHODS: Participants were examined by an ophthalmologist, who was unaware of their HIV-status. RESULTS: 238 participants were examined, 43 were HIV-1 prevalent cases (infected prior to 1990); 62 were HIV-1 incident cases and 133 were HIV-negative controls. Eleven of the HIV-positive participants had AIDS, and a further 8 had a CD4 count of less than 200 cells/mm3. Bilateral low vision was recorded in 9 participants and cataracts were the main cause of bilateral low vision. The mean intra-ocular pressure (IOP) was 12.6 mm Hg, and no association was found between IOP and CD4 lymphocyte count. Although significantly more HIV positives had some sign of intraocular inflammation compared to the negatives (p = 0.02) there was no significant linear trend in the HIV positives with decreasing CD4 count. Overall, 11 (26%) prevalent, 6 (10%) incident cases and 8 (6%) HIV-negatives controls had some evidence of intraocular inflammation, however, only one person had reduced visual acuity attributable to these lesions. Punctate epithelial erosions and keratic precipitates in the anterior chamber were seen significantly more frequently in HIV-positives compared to the HIV-negative controls. Cotton wool spots were not seen. CONCLUSIONS: A substantial proportion of HIV-infected persons had ocular findings which may have been attributable to their infection, however in only one case did this result in reduced visual acuity. Although the ocular complications of AIDS seem to comprise a large extra element in the work load of tertiary care hospitals dealing with eye problems, on a population basis such cases are infrequent.
Detection of myocardial viability in ischemic-reperfused rat hearts by Tc-99m sestamibi kinetics.
BACKGROUND: The purpose of this study was to evaluate technetium 99m sestamibi (MIBI) kinetics in assessing myocardial viability in hearts subjected to different ischemia-reperfusion treatments, resulting in graded severity of injury. METHODS AND RESULTS: Sixteen isolated Krebs-Henseleit-perfused rat hearts were divided into 3 groups: control (flow, 12 mL/min; n = 5), ischemic-reperfused with glucose (IR+G, n = 6), and ischemic-reperfused without glucose (IR-G, n = 5). MIBI (11.1 mBq [300 microCi]) was infused for 60 minutes (uptake), followed by a 60-minute clearance. MIBI uptake (percent injected dose per gram) was significantly decreased in the IR+G (2.07 +/- 0.31) and IR-G groups (2.03 +/- 0.23; P = not significant with IR+G) compared with the control group (3.06 +/- 0.25, P <.05). Fractional washout of MIBI was more rapid in the IR-G group (72.7% +/- 3.9%, P <.05) than in the control (21.9% +/- 1.9%) and IR+G groups (20.3% +/- 1.7%). End retention (percent injected dose per gram) of MIBI in the IR-G (0.60 +/- 0.12) and IR+G groups (1.60 +/- 0.18) was significantly less than in the control group (2.30 +/- 0.11, P <.05), respectively. The retention in the IR-G group was less than in the IR+G group (P <.05). Creatine kinase assay, triphenyltetrazolium chloride staining, and transmission electron microscopy analysis demonstrated more serious myocardial damage in the IR-G group than in the IR+G group. End MIBI activity was highly correlated with myocardial viability determined by triphenyltetrazolium chloride staining (r = 0.94; P <.05) and creatine kinase assay (r = -0.86; P <.05). CONCLUSIONS: Clearance of Tc-99m sestamibi is sensitive to metabolic states and may be used for assessment of ongoing myocardial damage.
Myocardial kinetics of Tc-99m glucarate in low flow, hypoxia, and aglycemia.
BACKGROUND: Technetium-99m glucarate is a myocardial infarct-avid imaging agent. Recent conflicting and inconclusive reports have suggested that the agent may be taken up by ischemic but viable myocardium. The purposes of this study were (1) to determine conclusively whether there is Tc-99m glucarate uptake in ischemic viable myocardium and (2) to investigate the potential mechanisms for such uptake by studying components of ischemia, namely, low flow, hypoxia, and aglycemia. METHODS AND RESULTS: Rat hearts were isolated and perfused in a modified Langendorff preparation with a crytalloid perfusate. Tc-99m glucarate was studied in control (n = 6), low-flow (n = 5), hypoxic (n = 5), and aglycemic (n = 5) conditions. The experimental protocol consisted of 20-minute baseline (12 mL/min flow) and 20-minute treatment (low flow at 1 mL/min, hypoxia, or aglycemia), followed by tracer uptake (20 minute) and washout (20 minutes). Activity was monitored with a sodium iodide detector. The tracer was delivered continuously over a 20-minute uptake period. The injected dose was 150 micro Ci (5.6 MBq). Hemodynamics were monitored throughout. Triphenyltetrazolium chloride staining was used to assess myocardial viability. There was no evidence of myocardial necrosis. Low flow tended to delay tracer uptake compared with control for the first 10 minutes, but this did not reach statistical significance. Low flow increased end fractional retention significantly compared with control (mean +/- SEM, 59.0% +/- 0.9% peak vs 41.2% +/- 1.4%, respectively; P <.05). Hypoxia resulted in a trend toward increased uptake; however, this was significant only at one early time point during the uptake phase. Retention in the hypoxia group was similar to control. Tc-99m glucarate uptake was significantly increased in aglycemia from 16 minutes to peak compared with control (1.36% +/- 0.71% injected dose per gram vs 0.91% +/- 0.37% injected dose per gram, respectively; P <.05). Aglycemia produced significantly higher end fractional retention compared with control (51.6% +/- 1.8% peak vs 41.2% +/- 1.4%, respectively; P <.05). CONCLUSIONS: Tc-99m glucarate myocardial retention is increased in the setting of ischemia, even in the absence of necrosis. This increased retention is not due to hypoxia. Furthermore, the retention is only partially explained by tissue hypoglycemia. Thus low flow per se appears to have a role in this increased retention, probably as a result of delayed flow-dependent washout.
Love and Haight: the sensuous hippie revisited. Drug/sex practices in San Francisco, 1980-81.
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Acupuncture as a detoxification modality.
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Venous aneurysms: MR diagnosis with the "layered gadolinium" sign.
OBJECTIVE: Our goal was to present MR findings in venous aneurysms and introduce the "layered gadolinium" sign as an ancillary diagnostic finding. METHOD: Gadolinium-enhanced MR images of three patients with retroperitoneal venous aneurysms were retrospectively reviewed. Prior to MRI, venous aneurysm had been suspected clinically in only one patient. Surgical correlation was available in one patient. A phantom was constructed and imaged to investigate the cause of the layered gadolinium sign. RESULTS: A gradation of signal intensity, the layered gadolinium sign, was observed in three patients with venous aneurysms on postcontrast T1-weighted images. The anterior portion of the aneurysms demonstrated high signal intensity separated by a sharp interface from the low signal intensity posterior region. Unenhanced time-of-flight MR venography, color Doppler, and duplex sonography failed to demonstrate flow in the patient with surgical proof. CONCLUSION: The layered gadolinium sign may be helpful in the diagnosis of venous aneurysm and in differentiating these masses from solid neoplasms.
A clinical study of hospital replacement mattresses.
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Inhibition by salicylate of gluconeogenesis in the isolated perfused rat liver.
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Facilitation of diabetes care in North Tyneside.
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Fat suppression in MR imaging: techniques and pitfalls.
Fat suppression is commonly used in magnetic resonance (MR) imaging to suppress the signal from adipose tissue or detect adipose tissue. Fat suppression can be achieved with three methods: fat saturation, inversion-recovery imaging, and opposed-phase imaging. Selection of a fat suppression technique should depend on the purpose of the fat suppression (contrast enhancement vs tissue characterization) and the amount of fat in the tissue being studied. Fat saturation is recommended for suppression of signal from large amounts of fat and reliable acquisition of contrast material-enhanced images. The main drawbacks of this technique are sensitivity to magnetic field nonuniformity, misregistration artifacts, and unreliability when used with low-field-strength magnets. Inversion-recovery imaging allows homogeneous and global fat suppression and can be used with low-field-strength magnets. However, this technique is not specific for fat, and the signal intensity of tissue with a long T1 and tissue with a short T1 may be ambiguous. Opposed-phase imaging is a fast and readily available technique. This method is recommended for demonstration of lesions that contain small amounts of fat. The main drawback of opposed-phase imaging is unreliability in the detection of small tumors embedded in fatty tissue.
Movement of dentinal and pulpal fluid caused by clinical procedures.
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