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Biomedical subjects

A M Taylor

Publications and source records attributed to A M Taylor.

At least 19 recordsLinked to original sources

Coronary artery imaging in grown up congenital heart disease: complementary role of magnetic resonance and x-ray coronary angiography.

BACKGROUND: There is a high incidence of anomalous coronary arteries in subjects with congenital heart disease. These abnormalities can be responsible for myocardial ischemia and sudden death or be damaged during surgical intervention. It can be difficult to define the proximal course of anomalous coronary arteries with the use of conventional x-ray coronary angiography. Magnetic resonance coronary angiography (MRCA) has been shown to be useful in the assessment of the 3-dimensional relationship between the coronary arteries and the great vessels in subjects with normal cardiac morphology but has not been used in patients with congenital heart disease. METHODS AND RESULTS: Twenty-five adults with various congenital heart abnormalities were studied. X-ray coronary angiography and respiratory-gated MRCA were performed in all subjects. Coronary artery origin and proximal course were assessed for each imaging modality by separate, blinded investigators. Images were then compared, and a consensus diagnosis was reached. With the consensus readings for both magnetic resonance and x-ray coronary angiography, it was possible to identify the origin and course of the proximal coronary arteries in all 25 subjects: 16 with coronary anomalies and 9 with normal coronary arteries. Respiratory-gated MRCA had an accuracy of 92%, a sensitivity of 88%, and a specificity of 100% for the detection of abnormal coronary arteries. The MRCA results were more likely to agree with the consensus for definition of the proximal course of the coronary arteries (P<0.02). CONCLUSIONS: For the assessment of anomalous coronary artery anatomy in patients with congenital heart disease, the use of the combination of MRCA with x-ray coronary angiography improves the definition of the proximal coronary artery course. MRCA provides correct spatial relationships, whereas x-ray angiography provides a view of the entire coronary length and its peripheral run-off. Furthermore, respiratory-gated MRCA can be performed without breath holding and with only limited subject cooperation.

Adult↗

A comparison between segmented k-space FLASH and interleaved spiral MR coronary angiography sequences.

A direct comparison of segmented fast low-angle short (FLASH) imaging and interleaved spiral magnetic resonance coronary angiography (MRCA) during free respiration using navigator echo has been performed. MRCA images were acquired in 30 normal subjects and 15 patients with coronary artery disease (CAD). Images of the right coronary artery were acquired during free respiration using navigator echo gating for both a segmented k-space FLASH sequence (8 views/segment, segment duration 105 msec) and an interleaved spiral sequence (20 interleaves, spiral read-out period 19 msec). Image quality was scored by three independent blinded observers, and coronary artery signal-to-noise ratio (SNR) and coronary artery/epicardial fat contrast-to-noise ratio (CNR) were measured. There was a significant improvement in image quality when coronary images were acquired with the interleaved spiral sequence (spiral 2. 3 vs. FLASH 1.8; P = 0.002). This was associated with an increase in the coronary artery SNR (16.6 +/- 6.9 vs. 11.8 +/- 5.0; P < 0.001), the coronary artery/epicardial fat CNR (12.5 +/- 6.1 vs. 7.4 +/- 4.0, P < 0.001), and the image resolution (256 x 256 vs. 256 x 128). However, there was a 12% increase in acquisition time for the interleaved spiral sequence. Image quality, SNR, CNR, and resolution can be improved using an interleaved spiral sequence. These improvements are secondary to the intrinsic characteristics of spiral imaging and the short acquisition period, which reduces the effects of both cardiac and respiratory motion.

Adult↗

Phase ordering with automatic window selection (PAWS): a novel motion-resistant technique for 3D coronary imaging.

Navigator acceptance imaging methods are hindered by the loss in scan efficiency which results from the changes in the breathing pattern of a subject over time. The diminishing variance algorithm (DVA), which does not use a predefined acceptance window, is less influenced by such changes. The use of phase ordering and weighting techniques has been shown to significantly improve image quality over nonordered window methods. However, the use of an acceptance window is inherent in all these techniques as a decision to accept or reject data must still be made. A technique is presented which is resistant to changes in breathing while allowing the use of phase ordering to provide effective motion artifact reduction in optimal time. The basic principle is described and illustrated for this automatic window-selection technique with in vitro results to demonstrate the feasibility of this method. Results of an in vivo study are also presented which demonstrate significant improvement in image quality over the DVA (p < 0.01) and hybrid-ordered phase encoding methods (p < 0.05).

Algorithms↗

Using language intensity to increase the success of a family intervention to protect children from ultraviolet radiation: predictions from language expectancy theory.

BACKGROUND: Even though people are informed about skin cancer prevention, they do not always comply with prevention advice. From Language Expectancy Theory, it was predicted that messages with high language intensity would improve compliance with sun safety recommendations and that this effect would be enhanced with deductive argument style. METHODS: Parents (N = 841) from a pediatric clinic and elementary schools received sun safety messages (newsletters, brochures, tip cards) by mail that varied in language intensity and logical style. Effects on attitudes and behavior were tested in a pretest-posttest factorial design. RESULTS: As hypothesized, parents receiving messages with high- as opposed to low-intensity language complied more with sun safety advice. Messages with highly intense language were more persuasive when the arguments were formatted in a deductive style; low language intensity was more persuasive in inductively styled messages. CONCLUSIONS: By carefully adjusting messages features, health professionals can obtain further compliance beyond that produced by educating people about health risks and creating favorable attitudes and self-efficacy expectations. Highly intense language may be a good general strategy in prevention messages and works better when conclusions and recommendations are offered explicitly to recipients, especially when advice is aimed at reducing their personal risk.

Assertiveness↗

Long-term effects of language intensity in preventive messages on planned family solar protection.

A series of sun safety messages containing highly intense language and deductive logical style achieved the most immediate compliance by parents, particularly when they intended to improve protection. Inductive messages were more successful when no intentions existed (D. B. Buller, Borland, & Burgoon, 1998; D. B. Buller et al., 2000). Interviewers recontacted 568 parents during the winter following message dissemination and assessed solar protection. A 2 (language intensity) x 2 (logical style) x 3 (behavioral intention) x 2 (person) mixed-model analysis of variance showed that parents receiving high-intensity, deductive messages reported the most improved solar protection and improvement was greatest when parents intended to improve protection. Over the long term, high language intensity may reinforce decisions to take preventive action and does not appear to provoke psychological reactance or resistance to these highly directive messages.

Analysis of Variance↗

Clonal diversity of Ig and T-cell receptor gene rearrangements in childhood B-precursor acute lymphoblastic leukaemia.

The majority of paediatric B precursor acute lymphoblastic leukaemias in children are derived from a single transformed haematopoietic cell with complete or partial VDJ recombination within the immunoglobulin heavy chain gene. A high frequency of patients also show rearrangements within TCRdelta and TCRgamma loci and in up to 40% of children there is an excess of immune system gene rearrangements compared with the number of identified alleles of immune system genes, suggesting the presence of multiple leukaemic subclones -clonal diversity. It has been observed by us and other investigators that in individual patients the pattern of immune system gene rearrangements often changes between presentation and relapse. In order to explore the possibility that clonal diversity plays a biological role during disease progression we optimised methods for subclone detection and analysed the prognostic significance of clonal diversity among 75 children with B precursor-ALL. Our results suggest that clonal diversity plays a role in disease progression as patients with oligoclonal disease showed a significantly shorter disease free survival than patients with monoclonal disease. This trend was of particular importance in the 'standard risk' group of ALL where aggressive disease could not be recognised by other means. In addition, generation of independent subclones from an early, non-rearranged tumour progenitor appears to be a common feature among leukaemias with aggressive clinical behaviour. We speculate on the type of genetic factors which may participate both in the generation of subclones and also in wider genomic instability and which are likely to be required for the aggressive clinical phenotype in children with ALL.

Burkitt Lymphoma↗

The DNA double-strand break repair gene hMRE11 is mutated in individuals with an ataxia-telangiectasia-like disorder.

We show that hypomorphic mutations in hMRE11, but not in ATM, are present in certain individuals with an ataxia-telangiectasia-like disorder (ATLD). The cellular features resulting from these hMRE11 mutations are similar to those seen in A-T as well as NBS and include hypersensitivity to ionizing radiation, radioresistant DNA synthesis, and abrogation of ATM-dependent events, such as the activation of Jun kinase following exposure to gamma irradiation. Although the mutant hMre11 proteins retain some ability to interact with hRad50 and Nbs1, formation of ionizing radiation-induced hMre11 and Nbs1 foci was absent in hMRE11 mutant cells. These data demonstrate that ATM and the hMre11/hRad50/Nbs1 protein complex act in the same DNA damage response pathway and link hMre11 to the complex pathology of A-T.

Acid Anhydride Hydrolases↗

Binding properties of the stilbene disulfonate sites on human erythrocyte AE1: kinetic, thermodynamic, and solid state deuterium NMR analyses.

A novel stilbene disulfonate, 4-trimethylammonium-4'-isothiocyanostilbene-2,2'-disulfonic acid (TIDS), has been chemically synthesized, and the interaction of this probe with human erythrocyte anion exchanger (AE1) was characterized. Covalent labeling of intact erythrocytes by [N(+)((14)CH(3))(3)]TIDS revealed that specific modification of AE1 was achieved only after removal of other ligand binding sites by external trypsinization. Following proteolysis, (1.2 +/- 0.4) x 10(6) TIDS binding sites per erythrocyte could be blocked by prior treatment with 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS), a highly specific inhibitor of AE1. Inhibition of sulfate equilibrium exchange by TIDS in whole cells was described by a Hill coefficient of 1.10 +/- 0.06, which reduced to 0.51 +/- 0.01 following external trypsinization. The negative cooperativity of TIDS binding following external trypsinization suggests that trypsin-sensitive proteins modulate allosteric coupling between AE1 monomers. Thermodynamic analysis revealed that TIDS binding induces smaller conformational changes in AE1 than is observed following DIDS binding. The similar inhibitory potencies of both TIDS (IC(50) = 0.71 +/- 0.48 microM) and DIDS (IC(50) = 0.2 microM) imply that there is no correlation between the ability of stilbene disulfonates to arrest anion exchange function and the magnitude of ligand-induced conformational changes in AE1. Solid state (2)H NMR analysis of a [N(+)(CD(3))(3)]TIDS-AE1 complex in both unoriented and macroscopically oriented membranes revealed that large amplitude "wobbling" motions describe ligand dynamics. The data are consistent with a model where TIDS bound to AE1 is located exofacially in contact with the bulk aqueous phase.

Anion Exchange Protein 1, Erythrocyte↗

Thiamine deficiency and malaria in adults from southeast Asia.

BACKGROUND: Thiamine deficiency (beriberi) is common in some parts of southeast Asia. Acute thiamine deficiency can mimic many complications of malaria, such as encephalopathy and lactic acidosis. We examined the incidence of thiamine deficiency in adults admitted to hospital with malaria in Thailand. METHODS: For this prospective study, we recruited consecutive patients with malaria or other febrile illness who presented to Paholpolpayuhasena Hospital, Kanchanaburi, Thailand, between May and July, 1992. We used the activation coefficient (alpha) for transketolase activity in erythrocytes to measure thiamine deficiency (defined as alpha>1.31) in patients with severe and uncomplicated malaria and in controls (patients' relatives and healthy volunteers). To exclude the possibility of interference in the assays, transketolase activity was also measured in erythrocytes used to culture parasites. FINDINGS: 12 (52%) of 23 patients with severe malaria and ten (19%) of 54 patients with uncomplicated malaria had alpha values above the normal range (p<0.0001 and p=0.0014, respectively, compared with controls), which indicated severe thiamine deficiency. Thiamine deficiency was more severe in patients with cerebral malaria than in those with uncomplicated malaria and the controls (p=0.008). INTERPRETATION: In adults admitted to hospital in Thailand, thiamine deficiency commonly complicates acute falciparum malaria, particularly in severe infections, and could contribute to dysfunction of the central nervous system.

Acidosis, Lactic↗

Inactivation of ataxia telangiectasia mutated gene in B-cell chronic lymphocytic leukaemia.

BACKGROUND: Patients with the inherited disorder ataxia telangiectasia (A-T) have an increased susceptibility to lymphoid malignancies. In these patients mutations affect both alleles of the A-T gene (ATM). We have looked for mutations in the ATM gene in sporadic cases of B-cell chronic lymphocytic leukaemia (B-CLL). METHODS: 32 cases of B-CLL were analysed by restriction endonuclease fingerprinting to detect mutations within ATM. In six of the cases in which mutations were detected in tumour samples, germline DNA was screened to assess ATM carrier status. The samples in 20 cases were also studied by western blot for abnormal expression of ATM protein. FINDINGS: Expression of the ATM protein was impaired in eight (40%) of the 20 tumours analysed, being absent in three and decreased in five. Mutations within ATM were detected in six (18%) of the 32 patients. These point mutations, deletions, and one insertion were distributed across the coding sequence of ATM. Germline mutations, which indicate ATM carrier status, were found in two of these six patients compared with a frequency within the general population of below 1 in 200. INTERPRETATION: Abnormal expression of ATM protein is a frequent finding in B-CLL. Although the precise function of this protein is unknown, it is thought to have a role in programmed cell death, a deficiency of which would fit with the characteristic phenotype of prolonged cell survival seen in B-CLL tumour cells. Our results also suggest that carriers of ATM mutations may be at a particular risk for the development of B-CLL and this may partly explain the known genetic susceptibility to this disease.

Alleles↗

Magnetic resonance navigator echo diaphragm monitoring in patients with suspected diaphragm paralysis.

Real-time magnetic resonance (MR) navigator echo (NE) monitoring of the diaphragm is now possible. Using this technique, temporal changes in diaphragm position can be analyzed in a non-invasive fashion, without x-ray exposure. In this preliminary study, we have optimized three NE parameters (the NE column area, the NE repeat time, and the location of the NE on the diaphragm surface), and demonstrated the clinical application of MR NE diaphragm monitoring in patients with suspected diaphragm paralysis. The NE parameters were defined in 10 healthy volunteers, and diaphragm traces were scored for variance in NE diaphragm position registration. Using the optimal NE column parameters, we investigated four patients with diaphragm paralysis, one of whom required positive pressure ventilation while in the MR scanner, to show the utility of this technique. The NE diaphragm position registration was significantly affected by the area of the NE column, with poor position registration for the smallest column area (2.25 cm2 vs. 4 cm2 vs. 6.25 cm2, variance 6.3 vs. 0.6 vs. 0.3, P = 0.006). Diaphragm position registration was also significantly affected by the NE repeat time, with misregistration for the shortest repeat time (250 msec vs. 500 msec vs. 1000 msec, variance 11.9 vs. 0.6 vs. 1.0, P = 0.02), and data clipping, with loss of end-expiratory and end-inspiratory position registration, for the longest repeat time. Finally, if the NE was positioned too anteriorly, the diaphragm traces were of poor quality (anterior vs. dome vs. posterior, variance 11.8vs. 0.6vs. 3.2, P < 0.001). Application of the technique confirmed diaphragm paralysis in all four patients. The technique can be applied during positive pressure ventilation if necessary. The optimal NE parameters for diaphragm monitoring at 0.5 T were: column area, 400 mm2; NE repeat time; 500 msec; NE column positioned on the diaphragm dome. MR NE diaphragm monitoring provides a safe, non-invasive method of assessing diaphragm motion in patients with suspected diaphragm paralysis and may prove useful for long-term follow-up and monitoring of therapeutic interventions in these subjects.

Adult↗

Safety and preliminary findings with the intravascular contrast agent NC100150 injection for MR coronary angiography.

In this Phase I clinical study, a novel ultrasmall superparamagnetic iron oxide contrast agent, NC100150 Injection (Nycomed Imaging, Oslo, Norway, a part of Nycomed Amersham), was used in two-dimensional magnetic resonance coronary angiography (MRCA). Safety and imaging data were acquired from 18 healthy male volunteers at both 0.5 and 1.5 T, before and after the administration of NC100150 Injection. Through-plane and in-plane images of the right coronary artery were analyzed. The postcontrast imaging sequences used prepulses and a high flip angle, to introduce T1 weighting. At 1.5 T (TE 2.6 msec), the through-plane coronary artery signal-to-noise ratio (SNR) (P = 0.04), coronary artery-to-fat signal difference-to-noise ratio (SDNR) (P = 0.001), coronary artery-to-myocardium SDNR (P<0.001), and coronary artery delineation (P<0.001) were improved by the administration of NC100150 Injection. For in-plane imaging, coronary artery delineation improved, but there were no significant changes in the SNR and SDNR. At 0.5 T, with the longer TE (6.7 msec) imaging sequence used, there was a reduction in the SNR (P = 0.01), the fat SDNR (through-plane P = 0.02; in-plane P = 0.25), and the coronary artery diameter (P<0.01 in both imaging planes). There was a trend toward improvement in the myocardial SDNR and coronary artery delineation. In conclusion, NC 100150 Injection was given safely to 18 healthy subjects, with no major adverse reactions. Coronary artery delineation was improved in both imaging planes at 1.5 T, with a trend toward improvement at 0.5 T. At 1.5 T, with a short TE imaging sequence, the marked T1 shortening effects of NC100150 Injection were dominant, leading to an improvement in the quantitative parameters for the through-plane images. At 0.5 T, with a longer TE imaging sequence, the T2* effects of the contrast agent played a role in reducing the quantitative image parameters. With further optimization of imaging sequences, to take advantage of the long-lived intravascular T1 shortening effect of NC100150 Injection, further improvements in MRCA will be possible.

Adult↗

Automated monitoring of diaphragm end-expiratory position for real-time navigator echo MR coronary angiography.

Real-time navigator echo (NE)-gated magnetic resonance coronary angiography (MRCA) during free respiration is now possible. However, the mean diaphragm end-expiratory position (DEEP) drifts over time, and this results in a reduction in scanning efficiency and increased artifacts due to the acquisition of data during periods of high diaphragm velocity. To address these problems, a diaphragm monitoring program that follows the mean DEEP over time has been developed. Fifteen subjects with ischemic heart disease underwent continuous NE monitoring of their diaphragm for 30 minutes. Using these diaphragm traces, theoretical MRCA scans were performed. Several diaphragm monitoring algorithms were developed and compared with the simplest case (a stationary 5 mm NE acceptance window placed around the mean DEEP, as measured by NE monitoring at the outset of the scan). An overall scan efficiency was calculated, and the number of completed scans where the mean DEEP lay within the NE acceptance window was recorded. Of the six algorithms considered, the most effective one monitored the mean DEEP and prospectively placed the upper limit of the NE acceptance window on this position for the subsequent acquisition. Using this algorithm in comparison with the simplest stationary scenario, both scan efficiency (47.9% vs. 38.5%, P = 0.01) and the number of completed scans where the mean DEEP lay within the NE acceptance window (71.2 vs. 30.3, P < 0.001) were improved. The implementation of such a monitoring algorithm, in combination with adaptive motion correction techniques, should improve overall scan efficiency while maintaining the end-expiratory position at the top end of the NE acceptance window, to reduce image artifacts.

Adult↗

Differences between normal subjects and patients with coronary artery disease for three different MR coronary angiography respiratory suppression techniques.

A comparison between three magnetic resonance coronary angiography (MRCA) respiratory motion suppression techniques was performed for both normal subjects and patients with coronary artery disease (CAD). MRCA images were acquired in 17 normal subjects and 15 patients with CAD, using conventional breath-hold MRCA, navigator echo (NE)-guided breath-hold MRCA (LED feedback), and NE-gated MRCA during free respiration. Image quality, diaphragm registration, and total acquisition time were assessed. Overall, there was poor diaphragm registration for conventional breath-holding compared with free respiration (P < 0.001). CAD patients found it significantly more difficult to perform a steady breath-hold (P = 0.04) or attain the same diaphragm position over multiple breath-holds than normal subjects (P = 0.02). All normal subjects, but only 3 of the 15 CAD patients, were able to perform the LED feedback technique (P < 0.001). For normal subjects, image quality was similar between the three respiratory suppression techniques (P = 0.3), while for CAD patients there was an improvement in image quality, for images acquired during free respiration (breath-hold vs. free respiration, P < 0.01). There was no significant difference in the total acquisition times between the breath-hold and free respiration techniques (P = 0.2). There were substantial differences in the effectiveness of MRCA respiratory suppression techniques between normal subjects and CAD patients. In patients, only NE-gated MRCA performed well, requiring minimal cooperation with no increase in total acquisition time. Validation of NE-MRCA techniques should always be performed in patients, as well as normal subjects, to ensure correct evaluation of the technique for the target population.

Adult↗

First-pass myocardial perfusion imaging and equilibrium signal changes using the intravascular contrast agent NC100150 injection.

In this phase I clinical study, the new ultrasmall superparamagnetic iron oxide contrast agent, NC100150 Injection (Nycomed AS, Oslo, Norway, a part of Nycomed Amersham), was assessed for first-pass magnetic resonance myocardial perfusion studies and its ability to produce equilibrium signal changes, as a possible indicator of myocardial blood volume. Data were acquired in 18 healthy male volunteers at 0.5 T and 1.5 T. At both field strengths, first-pass studies using T1-weighted sequences were acquired. Long TE spin-echo echoplanar imaging (EPI) was used at 0.5 T and short TE fast low-angle shot (FLASH) imaging at 1.5 T. With both sequences, T1 effects dominated the images for low doses, and time intensity curves potentially suitable for perfusion analysis were generated. At higher doses, T2 and T2* effects were observed. At 1.5 T, these predominantly affected the blood pool signal; however, at 0.5 T the myocardial signal was also involved, reflecting the relative T2 and T2* sensitivity of the spin-echo EPI sequence as a result of the long TE and long readout window, respectively. Equilibrium changes were assessed at both field strengths using T1-weighted FLASH sequences and in addition at 1.5 T using T2*-weighted gradient-echo EPI. With the T1-weighted images at both field strengths, signal changes were observed in all subjects; however, no dose-response relationship could be shown. With the T2*-weighted EPI there was significantly lower signal (P < 0.05) with the 3 and 4 mg/kg doses than with the 2 mg/kg dose. In conclusion, NC100150 Injection is useful for first-pass myocardial perfusion using T1-weighted sequences; however, low doses in combination with short TE sequences are required to minimize sensitivity to T2* effects. Equilibrium signal changes can also be induced in the myocardium. More work is required to optimize the imaging sequences and dose of NC100150 Injection for first-pass studies and also to determine whether the equilibrium signal changes can be used to measure myocardial blood volume changes in ischemic heart disease.

Adult↗

Coronary artery imaging in a 0.5-Tesla scanner: implementation of real-time, navigator echo-controlled segmented k-space FLASH and interleaved-spiral sequences.

Coronary angiography techniques have been implemented on a 0.5-Tesla scanner with a view to performing coronary artery imaging. Slice-followed, segmented k-space FLASH sequences and interleaved-spiral sequences have been employed with acquisitions under real-time navigator echo control with patient feed back, enabling poor signal-to-noise levels to be overcome by averaging data acquired over multiple, variable-length, reproducible breath holds. Good-quality, millimetre-resolution coronary images were obtained in ten normal subjects with both techniques. The mean percent of data segments or interleaves acquired with the navigator echo within the 5-mm diaphragm acceptance window was 57% [standard deviation (S.D.), 11%; range, 38-85%], and the average image-acquisition times were 123+/-22 sec and 71+/-14 sec for segmented FLASH and interleaved-spiral imaging, respectively. In addition to shorter acquisition times, the interleaved-spiral sequence has superior temporal resolution, allowing the acquisition of limited, multislice data sets. However, the sequence is particularly sensitive to the off-resonance effects of residual epicardial fat surrounding the artery and to field nonuniformities, both of which lead to image blurring and, unlike segmented FLASH acquisitions (which are very robust), the spiral data sets generally require postprocessing.

Adult↗

3D coronary artery imaging with phase reordering for improved scan efficiency.

Three-dimensional (3D) coronary imaging has the potential to overcome problems resulting from vessel tortuosity and to reduce partial volume effects. With these techniques, however, acquisition times are long and respiratory motion artifacts problematical. This work describes the development of a method that applies phase encode reordering to 3D acquisitions, allowing larger navigator acceptance windows to be used, with a consequent reduction in acquisition time. This method is compared with navigator acceptance window methods (the acceptance-rejection algorithm and the diminishing variance algorithm) and the retrospective respiratory gating technique, both in vitro and in vivo. The use of phase reordering with a 10 mm acceptance window provided a significant increase in scan efficiency over a non-reordered 5 mm method (P<0.001) with no significant change in image quality, and a significant increase in image quality compared with a non-reordered image acquired in the same time (P<0.05). A significant improvement in both image quality and scan efficiency was demonstrated over the retrospective respiratory gating method (P<0.05).

Adult↗

Hydrodynamic properties of human erythrocyte band 3 solubilized in reduced Triton X-100.

The oligomeric state and function of band 3, purified by sulfhydryl affinity chromatography in reduced Triton X-100, was investigated. Size exclusion high-performance liquid chromatography showed that a homogeneous population of band 3 dimers could be purified from whole erythrocyte membranes. The elution profile of band 3 purified from membranes that had been stripped of its cytoskeleton before solubilization was a broad single peak describing a heterogeneous population of oligomers with a mean Stokes radius of 100 A. Sedimentation velocity ultracentrifugation analysis confirmed particle heterogeneity and further showed monomer/dimer/tetramer equilibrium self-association. Whether the conversion of dimer to the form described by a Stokes radius of 100 A was initiated by removal of cytoskeletal components, alkali-induced changes in band 3 conformation, or alkali-induced loss of copurifying ligands remains unclear. After incubation at 20 degrees C for 24 h, both preparations of band 3 converted to a common form characterized by a mean Stokes radius of 114 A. This form of the protein, examined by equilibrium sedimentation ultracentrifugation, is able to self-associate reversibly, and the self-association can be described by a dimer/tetramer/hexamer model, although the presence of higher oligomers cannot be discounted. The ability of the different forms of the protein to bind stilbene disulfonates revealed that the dimer had the highest inhibitor binding affinity, and the form characterized by a mean Stokes radius of 114 A to have the lowest.

Anion Exchange Protein 1, Erythrocyte↗