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Biomedical subjects

W C Chu

Publications and source records attributed to W C Chu.

At least 19 recordsLinked to original sources
PubMed2005

Accuracy of endoscopic ultrasonography in diagnosing ascites and predicting peritoneal metastases in gastric cancer patients.

BACKGROUND: Preoperative diagnosis of peritoneal metastases (PM) is difficult in patients with gastric cancer (GC). AIMS: To assess the accuracy of endoscopic ultrasonography (EUS) in diagnosing ascites and its predictability for the presence of PM in GC patients. SUBJECTS: Consecutive patients with newly diagnosed GC from 1998 to 2004 were studied. METHODS: All patients underwent EUS, ultrasound (US), and computed tomography (CT) scan for preoperative staging and the presence of ascites. The results were compared with operative findings. The diagnosis of PM was confirmed by histopathology or peritoneal fluid cytology. RESULTS: A total of 301 patients were recruited and in 250 patients the presence of ascites (n = 93) and PM (n = 71) were confirmed. EUS was more sensitive (87.1%) than combined US and CT scan examinations (16.1%) and operative findings (laparoscopy or laparotomy) (40.9%) in diagnosing ascites. Sensitivity, specificity, positive and negative predictive values, and accuracy for predicting the presence of PM were 73%, 84%, 64%, 89%, and 81% by EUS; 18%, 99%, 87%, 75%, and 76% by combining US and CT scan; and 77%, 94%, 83%, 91%, and 89% by operative findings, respectively. In multivariate logistic regression analysis, EUS detected ascites was the only significant independent predictor for the presence of PM (p<0.001; odds ratio 4.7 (95% confidence interval 2.0-11.2)). CONCLUSION: EUS is a sensitive method for diagnosing ascites which is an important predictive factor for the presence of PM in GC patients.

Adenocarcinoma↗
PubMed2005

A least-squares error minimization approach in the determination of ferric ion diffusion coefficient of Fricke-infused dosimeter gels.

A least-squares error minimization approach was adopted to assess ferric ion diffusion coefficient of Fricke-agarose gels. Ferric ion diffusion process was modeled as a Gaussian-shaped degradation kernel operating on an initial concentration distribution. Diffusion coefficient was iteratively determined by minimizing the error function defined as the difference between the theoretically calculated and the experimentally measured dose distributions. A rapid MR image-based differential gel dosimetry technique that time resolves the evolution of the ferric ion diffusion process minimizes smearing of the dose distribution. Our results showed that for a Fricke-agarose gel contained 1 mM ammonium ferrous sulfate, 1% agarose, 1 mM sodium chloride, and 50 mM sulfuric acid, its ferric ion diffusion coefficient is (1.59 +/- 0.28) x 10(-2) cm2 h(-1) at room temperature. This value falls within the 1.00-2.00 x 10(-2) cm2 h(-1) range previously reported under varying gelling ingredients and concentrations. This method allows a quick, nondestructive evaluation of the ferric ion diffusion coefficient that can be used in conjunction with the in situ gel dosimetry experiment to provide a practical diffusion characterization of the dosimeter gel.

Diffusion↗
PubMed2002

The role of dose distribution gradient in the observed ferric ion diffusion time scale in MRI-Fricke-infused gel dosimetry.

Ferric ion diffusion is a detrimental factor in MRI-Fricke-infused gel dosimetry. In this study, a novel approach involving MR image subtraction and a fast image-based dosimetry technique to study ferric ion diffusion effects is presented. The fast image-based approach allows studying dose profile degradation within minutes post-irradiation. The relationship between the rate of dose profile deterioration and dose distribution gradients can be elucidated with the improved imaging temporal resolution also. Our results showed that for a dose distribution with gradient 4 Gy/mm or higher, ferric ion diffusion causes apparent dose profile degradation in 0.5-1 h post-irradiation. For a gradual dose gradient change of 2.1 Gy/mm or smaller, dose profile degradation appears insignificant for a two-hour elapsed diffusion time. These observations agree well with the theoretical analysis of a square dependence between dose profile degradation and dose distribution gradient. Because all stereotactic radiosurgery procedures produce steep dose distributions and because the ideal "snapshot" of MR scanning cannot be achieved, knowledge of the ferric ion diffusion time scale is important in experimental designs in order to avoid potential measurement errors in MRI-Fricke-agarose gel dosimetry.

Diffusion↗
PubMed2001

In vivo magnetic resonance (MR) study of fatty liver: importance of intracellular ultrastructural alteration for MR tissue parameters change.

Fatty liver is thought to have a shorter T1 relaxation time than normal liver tissue, due to the accumulation of triglyceride. Previous studies regarding T1 and T2 relaxation times, however, show widely different results. In order to elucidate the mechanism responsible for the changes and diversity of relaxation times in fatty liver, we created two animal models in 14 rabbits, one acute form (N = 6) and the other chronic form (N = 8). Four rabbits were taken as a control group. Tissue relaxation times and the magnetization transfer (MT) effect of the liver tissue in these two models were measured. The results were correlated with biochemical analysis of water and fat content and histological examination, including findings in light microscopy and electron microscopy. Although the fatty ratio in both forms of fatty liver was similar, their tissue relaxation rate and MT effect were significantly different. The acute form showed prolongation of both T1 and T2 relaxation times (512 +/- 51 msec vs. 710 +/- 95 msec and 39 +/- 1.8 msec vs. 48 +/- 3.7 msec, respectively) and a decrease of the MT effect (50 +/- 5.1% vs. 38 +/- 6.3%), compared to those of the control group and preinduction liver. The chronic form showed shorter T1 and T2 values (526 +/- 36 msec vs. 406 +/- 56 msec and 36 +/- 1.6 msec vs. 33 +/- 2.3 msec, respectively) and a stronger MT effect (21 +/- 0.9% vs. 26 +/- 2.3%). In acute form fatty liver, electron microscopic examination revealed dramatic subcellular changes, such as vesicular transformation, a markedly increased amount of smooth endoplasmic reticulum (SER), and disruption of the crista. These changes were not found in the chronic form fatty liver. From this study, we concluded that the ultrastructural alteration in the subcellular organelles of hepatocyte might play a crucial role for the chameleonic presentation of MR tissue parameters in fatty liver.

Animals↗
PubMed2001

Improving relative anisotropy measurement using directional correlation of diffusion tensors.

A method employing directional correlation of the diffusion tensor, directional-correlation weighted relative anisotropy (DRA), was developed to improve the accuracy of estimated relative anisotropy (RA). The intravoxel directional correlation was established on the same voxel between two identically acquired diffusion tensor images, and the correlation coefficient derived from tensor dot product was employed as the weighting factor applied in the calculation of RA. The effect of noise influence was reduced since the random noise between repeated scans is not directionally correlated. The RA and the inter- and intravoxel DRA estimations were examined on rat brains in vivo. The background noise alters the direction of eigenvectors and the magnitude of eigenvalues. The dispersion angle between repeatedly obtained eigenvectors, representing the extent of directional alteration of eigenvectors, depends on the tissue anisotropy as well as the signal-to-noise ratio (SNR) of the source images. Current results demonstrate that the intravoxel DRA improves the accuracy of RA estimation, increases the relative contrast of gray and white matter, and avoids the partial volume effect commonly seen in the intervoxel operations.

Animals↗
PubMed2001

Video cystometry in young infants with renal dilation or a history of urinary tract infection.

Cystometry is increasingly being used in infants to diagnose bladder dysfunction. However, infantile urodynamic patterns have not been fully established. In this study we investigated the urodynamic patterns in young infants with renal dilation or a history of urinary tract infection, but with no apparent lower urinary tract symptoms. We use video cystometry with simultaneous perineal EMG recording. Thirty-five infants (27 male and 8 female) with congenital renal dilatation or a history of urinary tract infection at age 2 days to 24 months old were involved. We found that detrusor instability occurred in 8.6% of these subjects. Bladder capacity increased with age but less than would nomally be expected. An intermittent voiding pattern was observed in 57% (20/35) of subjects and was characterized by a single or recurring increase in sphincter activity with a simultaneous rise in the voiding detrusor pressure curve. The maximum voiding detrusor pressure with pelvic floor overactivity was significantly higher than that with no pelvic floor overactivity (105+/-44 cmH2O vs 69+/-22 cmH2O, P < 0.001). The median post-voiding residual volume was 2 (range 0 to 65) ml. We conclud that in infants with no apparent lower urinary tract symptoms, bladder instability is uncommon, and the capacity is lower than the normally expected range; an intermittent voiding pattern is common and the residual urine volume showed great variation. This probably represents an immature detrusor-sphincter function.

Dilatation, Pathologic↗
PubMed2001

Radiation dosimetry using Fricke-infused gels and magnetic resonance imaging.

We have witnessed the advancements of MRI-Fricke-infused gel dosimetry since its commencing in 1984. Over the years, many efforts have been spent to improve the method's efficacy, i.e., to improve its dose-response sensitivity, reproducibility and measurement accuracy. In this article, we give a review of the development of this relatively new dosimetric method. An example of applying this method to gamma knife stereotactic radiosurgery dose distribution mapping is also given.

Cations↗
PubMed2000

Incidence of adverse events after I.V injection of MR contrast agents in a Chinese population. A comparison between gadopentetate and gadodiamide.

PURPOSE: To study the incidence of adverse events after i.v. injection of MR contrast agents in a Chinese population. A comparison was made between an ionic contrast agent (dimeglumine gadopentetate, Magnevist) and a non-ionic contrast agent (gadodiamide, Omniscan). MATERIAL AND METHODS: During a 24-month period, 2,049 Chinese patients who randomly received an i.v. bolus injection of either Magnevist or Omniscan were investigated. All patients were questioned for the presence of any generalized or localized adverse reaction on the following day after the MR examination according to a standardized questionnaire. RESULTS: Three hundred and nine out of 2,049 patients (15%) reported an adverse event. There was a higher incidence of adverse events in patients receiving Magnevist as compared to those receiving Omniscan injection. All reported adverse events were clinically mild and required neither treatment nor hospitalization. CONCLUSION: There was a higher incidence of adverse reaction in patients receiving Magnevist than in those receiving Omniscan.

Adolescent↗
PubMed2000

Deferoxamine-induced bone dysplasia in the distal femur and patella of pediatric patients and young adults: MR imaging appearance.

OBJECTIVE: We investigated the MR imaging appearance of deferoxamine-induced bone dysplasia in the distal femur and patella in patients with thalassemia major. MATERIALS AND METHODS: Thirty-five patients with homozygous ss-thalassemia major who were undergoing regular transfusions and chelation therapy underwent coronal T1-weighted MR imaging of the femur, including the femoral head and the distal femoral epiphysis. Additional coronal fat-saturated dual-echo and sagittal T1-weighted images of the distal femur and patella were obtained in 11 patients who were suspected of having distal femoral lesions on the basis of the coronal T1-weighted images of the entire femur. RESULTS: No dysplastic change was detected in the proximal femur on coronal T1-weighted images. In 22 distal femurs of 11 patients, the following abnormalities were detected on MR imaging: blurred physeal-metaphyseal junction (n = 22), distal metaphyseal areas of hyperintensity (n = 21), physeal widening (n = 18), metadiaphyseal lesions (n = 11), epiphyseal lesions (n = 10), and patellar lesions (n = 2). Physeal widening and distal metaphyseal hyperintense areas were all more pronounced peripherally. Of the 21 distal metaphyseal hyperintensities, lateral abnormalities were larger than medial abnormalities in 16. Of the 18 distal femurs in which physeal widening was detected, the lateral widening was more marked than the medial widening in 12. Patients with MR imaging evidence of bone dysplasia have a significantly (p = 0.003) greater height reduction than patients without such evidence of bone dysplasia. CONCLUSION: Deferoxamine-induced bone dysplasia in the distal femur and patella is represented by a spectrum of morphologic changes in the epiphysis, physis, metaphysis, and metadiaphysis on MR imaging.

Adolescent↗
PubMed1999

Synthetase recognition determinants of E. coli valine transfer RNA.

We have studied the interactions between Escherichia coli tRNAVal and valyl-tRNA synthetase (ValRS) by enzymatic footprinting with nuclease S1 and ribonuclease V1, and by analysis of the aminoacylation kinetics of mutant tRNAVal transcripts. Valyl-tRNA synthetase specifically protects the anticodon loop, the 3' side of the stacked T-stem/acceptor-stem helix, and the 5' side of the anticodon stem of tRNAVal against cleavage by double- and single-strand-specific nucleases. Increased nuclease susceptibility at the ends of the anticodon- and T-stems in the tRNAVal.ValRS complex is indicative of enzyme-induced conformational changes in the tRNA. The most important synthetase recognition determinants are the middle and 3' anticodon nucleotides (A35 and C36, respectively); G20, in the variable pocket, and G45, in the tRNA central core, are minor recognition elements. The discriminator base, position 73, and the anticodon stem also are recognized by ValRS. Replacing wild-type A73 with G73 reduces the aminoacylation efficiency more than 40-fold. However, the C73 and U73 mutants remain good substrates for ValRS, suggesting that guanosine at position 73 acts as a negative determinant. The amino acid acceptor arm of tRNAVal contains no other synthetase recognition nucleotides, but regular A-type RNA helix geometry in the acceptor stem is essential [Liu, M., et al. (1997) Nucleic Acids Res. 25, 4883-4890]. In the anticodon stem, converting the U29:A41 base pair to C29:G41 reduces the aminoacylation efficiency 50-fold. This is apparently due to the rigidity of the anticodon stem caused by the presence of five consecutive C:G base pairs, since the A29:U41 mutant is readily aminoacylated. Identity switch experiments provide additional evidence for a role of the anticodon stem in synthetase recognition. The valine recognition determinants, A35, C36, A73, G20, G45, and a regular A-RNA acceptor helix are insufficient to transform E. coli tRNAPhe into an effective valine acceptor. Replacing the anticodon stem of tRNAPhe with that of tRNAVal, however, converts the tRNA into a good substrate for ValRS. These experiments confirm G45 as a minor ValRS recognition element.

Anticodon↗
PubMed1999

Potential utility of MRI in the evaluation of children at risk of renal scarring.

BACKGROUND: Gadolinium-enhanced MRI has recently been employed in the diagnosis of acute pyelonephritis. Its potential utility in the diagnosis of renal scars in children is unknown. Objective. To evaluate the potential utility of MRI using fat-saturated T1-weighted (T1-W) and post-gadolinium, short-tau inversion-recovery (STIR) sequences in detecting renal scarring by comparison with technetium dimercaptosuccinic acid ((99 m)Tc-DMSA) renal scintigraphy in children at risk of renal scarring. MATERIALS AND METHODS: A group of 24 children with spina bifida and neurogenic bladder or anorectal anomaly was studied. No patient had a history of acute pyelonephritis. Documented urinary tract infection (UTI) was present in 10 children (42 %). The remaining 14 (58 %) children had a history of asymptomatic bacteriuria. None had clinical signs or symptoms of acute UTI at the time of the study. (99 m)Tc-DMSA and MRI were performed to detect renal scarring. (99 m)Tc-DMSA scans were supplemented with pinhole imaging. MRI of the kidneys employed a fat-saturated T1-W sequence and a post-gadolinium STIR sequence employing a short echo time. RESULTS: Of the kidneys studied, 33 % (n = 16) had evidence of a renal parenchymal defect suggestive of scarring on (99 m)Tc-DMSA. The concordance in the detection of a scarred kidney by post-gadolinium STIR sequence and (99 m)Tc-DMSA is 94 %; that by fat-saturated T1-W sequence and (99 m)Tc-DMSA is 82 %; that by both sequences (positive result on either sequence) and (99 m)Tc-DMSA is 100 %. Using (99 m)Tc-DMSA as the gold standard, MRI had a sensitivity of 100 % and a specificity of 78 % in the diagnosis of a scarred kidney. The concordance in the detection of a scarred zone by post-gadolinium STIR sequence and (99 m)Tc-DMSA is 68 %; that by fat-saturated T1-W sequence and DMSA is 44 %; that by both sequences (positive result on either sequence) and (99 m)Tc-DMSA is 84 %. MRI had a sensitivity of 84 % and a specificity of 86 % in the diagnosis of a scarred zone, using (99 m)Tc-DMSA as the gold standard. CONCLUSION: The detection rate for renal scarring on MRI using the fat-saturated T1-W and post-gadolinium STIR sequences is comparable to planar (99 m)Tc-DMSA. MRI is of potential utility in the evaluation of children at risk of renal scarring.

Anal Canal↗
PubMed1999

A post-processing technique for displaying vessels from routine fast-spin-echo images: MRI-derived angiography.

Fast-spin-echo magnetic resonance (MR) images are routine components of a standard MR brain examination. On these images, blood vessels are visible as black flow void. We report that by applying an enhancement filter to a stack of routine fast-spin-echo MR images, projected angiographic images can be generated. The vascular detail in the projected image is similar to that observed in a phase-contrast image. In addition to its advantage in obtaining vessel information from routine images, the proposed post-processing technique is fast, easy to implement and completely automatic. These images provide additional vessel information that is useful when MR angiography is unavailable or as an aid in planning dedicated MR angiographic studies.

Brain↗
PubMed1999

Effects of dietary omega3 and omega6 lipids and vitamin E on proliferative response, lymphoid cell subsets, production of cytokines by spleen cells, and splenic protein levels for cytokines and oncogenes in MRL/MpJ-lpr/lpr mice.

omega3 Fatty acid rich fish oil (FO) and vitamin E may delay the progress of certain autoimmune diseases. The present study examined the mechanisms of action of omega3 lipids and vitamin E in autoimmune-prone MRL/lpr mice suffering from extensive lymphoproliferation, lupus-like symptoms, and accelerated aging. To determine whether the effects of omega3 lipids in autoimmune disease is linked to vitamin E levels, weanling female MRL/lpr and congenic control MRL/++ mice were fed diets containing 10% corn oil (CO) or 10% FO at two levels of vitamin E (75 IU or 500 IU/kg diet) for 4 months. The appearance of lymph nodes was delayed in the mice fed FO, and higher levels of FO offered further protection against the appearance of lymph nodes. Analysis of the spleen cells revealed that the cells positive for Thy.1 and Fas were significantly higher in the MRL/++ mice. The groups fed high levels of vitamin E generally exhibited higher levels of Fas. The proliferative response of splenocytes of MRL/++ mice to mitogens was significantly higher compared with MRL/lpr mice. Interleukin (IL)-10 production by spleen cells was significantly higher in FO-fed MRL/lpr mice than in CO-fed mice. In mice fed a high level of vitamin E, the production of IL-12 and tumor necrosis factor-alpha was significantly lower and IL-2 was significantly higher than in animals fed a low level of vitamin E. Proinflammatory cytokines were higher in the MRL/lpr mice and both FO and vitamin E lowered the levels of proinflammatory cytokines and lipid mediators. Western blots revealed that c-myc and c-ras were significantly lower and IL-2 and transforming growth factor (TGF)-beta1 levels were significantly higher in the spleens of MRL/++ mice. FO lowered c-myc and high levels of vitamin E in the diets normalized the levels of TGF-beta1 in MRL/lpr mice. The observations from this study suggest that both FO and vitamin E modulate the levels of specific cytokines, decrease the levels of proinflammatory cytokines, inflammatory lipid mediators, and c-myc, and increase TGF-beta1 levels in spleens of MRL/lpr mice and thus may delay the progress of autoimmune diseases.

Journal Article↗
PubMed1999

Effects of dietary omega-3 and omega-6 lipids and vitamin E on serum cytokines, lipid mediators and anti-DNA antibodies in a mouse model for rheumatoid arthritis.

OBJECTIVE: Omega-3 (omega-3) fatty acid rich-fish oil (FO) and vitamin E (vit-E) may delay the progress of certain autoimmune diseases. The present study examined the mechanism of action of omega-3 and omega-6 lipids and vit-E on the serum cytokines and lipid mediators in autoimmune-prone MRL/lpr mice (a model for rheumatoid arthritis, RA). The lpr (lymphoproliferative) gene is overexpressed in these mice causing extensive lymphoproliferation, lupus-like symptoms and accelerated aging. METHODS: Weanling female MRL/lpr and congenic control MRL/++ mice were fed 10% corn oil (CO, omega6) or FO-based semipurified diets containing two levels of vitamin E (vit-E-75, I.U. and vit-E-500 I.U./Kg diet) for four months. At the end of the experiment, serum anti-DNA antibodies, cytokines and lipid mediators levels were determined. RESULTS: The appearance of enlarged lymph nodes was delayed in the mice fed FO, and the FO-500 IU vit-E diet offered further protection against enlargement of lymph nodes. The MRL/lpr mice exhibited significantly higher levels of serum anti-dsDNA antibodies. The FO-fed mice had significantly lower serum IL-6, IL-10, IL-12, TNF-alpha, PGE2, TXB2 and LTB4 levels compared with CO-fed mice. In mice fed 500 IU vit-E diets, the serum IL-6, IL-10, IL-12 and TNF-alpha levels were significantly lower and serum IL-1beta was significantly higher compared to 75 IU-vit-E-fed mice in CO/FO or both. The levels of anti-DNA antibodies, IL-4, IL-6, TNF-alpha, IL-10 and IL-12 were higher in the sera of MRL/lpr mice. The FO diet lowered the levels of these cytokines (except IL-4) and lipid mediators. Adding 500 IU of vit-E to the FO diet further lowered the levels of IL-6, IL-10, IL-12, and TNF-alpha. CONCLUSION: It is clear from our observations that the beneficial effects of FO can be enhanced by the addition of 500 IU of vit-E in the diet. The FO diet containing 500 IU of vit-E may specifically modulate the levels of IL-6, IL-10, IL-12 and TNF-alpha and thereby may delay the onset of autoimmunity in the MRL/lpr mouse model. The observations from this study may form a basis for selective nutrition intervention based on specific fatty acids and antioxidants in delaying the progress of RA.

Animals↗
PubMed1998

The radiation induced magnetic resonance image intensity change provides a more efficient three-dimensional dose measurement in MRI-Fricke-agarose gel dosimetry.

A detailed methodology has been developed to map the spatial dose distribution in a Fricke-agarose gel based on the radiation induced image intensity change in the gel's magnetic resonance (MR) images. Besides the linear correlation between the change in the gel's spin-lattice relaxation rate and the absorbed dose, it is shown here that the radiation induced image intensity change for T1-weighted spin-echo images with TE << TR correlates exponentially to the absorbed dose. Furthermore, at the lower dose region (< 15 Gy), the correlation is fairly linear and its sensitivity is high. The minimum detectable dose is shown to be equivalent to the one obtained using the conventional R1-based approach. Since only one T1-weighted image is required for the dose evaluation, compared to the R1-based method, the total MR imaging time can be reduced from hours to a few minutes. This extensive time reduction avoids ferric ion diffusion effects and provides a practical way to simply and effectively measure the three-dimensional dose distribution using the Fricke-agarose dosimeter gel.

Gels↗
PubMed1997

Role of acceptor stem conformation in tRNAVal recognition by its cognate synthetase.

Although the anticodon is the primary element in Escherichia coli tRNAValfor recognition by valyl-tRNA synthetase (ValRS), nucleotides in the acceptor stem and other parts of the tRNA modulate recognition. Study of the steady state aminoacylation kinetics of acceptor stem mutants of E.coli tRNAValdemonstrates that replacing any base pair in the acceptor helix with another Watson-Crick base pair has little effect on aminoacylation efficiency. The absence of essential recognition nucleotides in the acceptor helix was confirmed by converting E.coli tRNAAlaand yeast tRNAPhe, whose acceptor stem sequences differ significantly from that of tRNAVal, to efficient valine acceptors. This transformation requires, in addition to a valine anticodon, replacement of the G:U base pair in the acceptor stem of these tRNAs. Mutational analysis of tRNAValverifies that G:U base pairs in the acceptor helix act as negative determinants of synthetase recognition. Insertion of G:U in place of the conserved U4:A69 in tRNAValreduces the efficiency of aminoacylation, due largely to an increase in K m. A smaller but significant decline in aminoacylation efficiency occurs when G:U is located at position 3:70; lesser effects are observed for G:U at other positions in the acceptor helix. The negative effects of G:U base pairs are strongly correlated with changes in helix structure in the vicinity of position 4:69 as monitored by19F NMR spectroscopy of 5-fluorouracil-substituted tRNAVal. This suggests that maintaining regular A-type RNA helix geometry in the acceptor stem is important for proper recognition of tRNAValby valyl-tRNA synthetase.19F NMR also shows that formation of the tRNAVal-valyl-tRNA synthetase complex does not disrupt the first base pair in the acceptor stem, a result different from that reported for the tRNAGln-glutaminyl-tRNA synthetase complex.

Anticodon↗
PubMed1997

Localization of the major ethidium bromide binding site on tRNA.

Binding of ethidium bromide to Escherichia coli tRNAVal and an RNA minihelix based on the acceptor stem and T-arm of tRNAVal was investigated by 19F and 1H NMR spectroscopy of RNAs labeled with fluorine by incorporation of 5-fluorouracil. Ethidium bromide selectively intercalates into the acceptor stem of the tRNAVal. More than one ethidium bromide binding site is found in the acceptor stem, the strongest between base pairs A6:U67 and U7:A66. 19F and 1H spectra of the 5-fluorouracil-substituted minihelix RNA indicate that the molecule exists in solution as a 12 base-paired stem and a single-stranded loop. Ethidium bromide no longer intercalates between base pairs corresponding to the tRNAVal acceptor stem in this molecule. Instead, it intercalates between base pairs at the bottom of the long stem-loop structure. These observations suggest that ethidium bromide has a preferred intercalation site close to the base of an RNA helical stem.

Base Sequence↗