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PubMed2004

Measurement of 15N csa/dipolar cross-correlation rates by means of Spin State Selective experiments.

We propose a method for the determination of (15)N csa/dipolar cross-correlation rates based on the measurement of the two apparent transverse (or longitudinal) relaxation rates associated with each component of the nitrogen doublet (N(alpha) and N(beta)). This is achieved by inserting a spin state selective scheme in conventional inverse Carr-Purcell-Meiboom-Gill (or inversion-recovery) pulse sequence which allows for the edition of a HSQC-type spectrum for each of the spin states. Transverse cross-correlation rates necessitate two independent sets of measurements (for N(alpha) and N(beta), respectively), whereas for longitudinal cross correlation rates, besides N(alpha) and N(beta) measurements, the method requires the knowledge of both the (15)N longitudinal auto-relaxation rate and the longitudinal two-spin order (2NzHz) auto-relaxation rate. These additional parameters are mandatory because of the non-exponential behavior of the N(alpha) and N(beta) longitudinal decays. Conversely, the present method does not require any complex manipulation of 2D spectra, the cross-correlation rates being obtained from the difference of the two (N(alpha) and N(beta)) apparent relaxation rates. This approach is applied to (15)N-labelled ubiquitin at two different magnetic fields (9.4 T and 14.1 T).

Magnetics↗
PubMed1981

Levels of processing in speech production.

When subjects repeated short alphabetic sequences at experimenter-controlled rates while retaining consonant trigrams, subsequent recall was unrelated to speech rate if single letters or a long sequence was repeated, but recall was inversely related to speech rate for sequences three or five letters in length. The latter effect was predicted by a model in which sequences were organized as single response units in a central phonetic processor but independently executed by means of mechanisms not requiring the services of the central processor. Quantitative predictions were generated from this model by estimating organization time for the sequences and assuming that the probability of forgetting would be equal to the organization time/execution time ratio. All obtained recall probabilities were as predicted for sequences three of five letters in length with both experimenter- and subject-controlled speech rates. Repetition of a nine-letter sequence from the end of the alphabet produced forgetting patterns clearly out of accord with any organization time/execution time ratio, but a similar sequence from the beginning of the alphabet produced evidence of being a marginal case with unstable organization.

Humans↗
PubMed2001

Serial STIR magnetic resonance imaging correlates with clinical score of activity in thyroid disease.

PURPOSE: To assess the correlation between inflammatory activity in extraocular muscles measured with serial short tau inversion recovery (STIR) sequence magnetic resonance imaging (MRI) scans and clinical disease activity in thyroid eye disease. METHODS: In this retrospective study, 22 patients with thyroid eye disease who had undergone serial MRI scans using the STIR sequence were assessed. The signal intensity ratio (SIR) of the most inflamed extraocular muscle was compared with the Mourits score (a clinical measure of thyroid eye disease activity). RESULTS: The SIR value has previously been shown to correlate with clinical activity of thyroid eye disease. In a particular patient the SIR value increases in proportion with the clinical features of the disease as assessed by the Mourits rating system. When the change in STIR sequence MRI is compared with the change in Mourits score rating for a given patient the correlation is highly significant (p < 0.001). CONCLUSIONS: This study confirms that serial STIR sequence MRI scans correlate with clinical disease activity. Inflammatory activity in the most inflamed muscle (as measured by SIR) reflects overall disease activity. SIR values obtained from MRI scans provide a useful measure of disease activity and may help in the monitoring and treatment of this condition.

Female↗
PubMed2001

Operative results without invasive monitoring in patients with frontal lobe epileptogenic lesions.

PURPOSE: To further explore the still controversial issues regarding whether all or most candidates for epilepsy surgery should be investigated preoperatively with invasive long-term video-EEG monitoring techniques (ILTVE). METHODS: We studied five patients with intractable seizures since early childhood using the same protocol: clinical evaluation, magnetic resonance imaging (MRI) with fluid-attenuated inversion recovery (FLAIR) sequences, long-term video-EEG (LTVE) monitoring with scalp electroencephalogram (EEG), interictal single photon emission computed tomography (SPECT), positron emission tomography (PET), and neuropsychological testing. The patients' seizures had clinical features suggesting a frontal lobe (FL) origin. MRI scans revealed focal cortical dysplasia (CD) in four patients and a probable gliotic lesion in the fifth. The findings in both PET and SPECT images were congruent with those of the MRI. Scalp LTVE failed to localize the ictal onset, although the data exhibited features suggestive of both CDs and FL seizures. On the basis of these results, surgery was performed with intraoperative corticography, and the cortical area exhibiting the greatest degree of spiking was ablated. RESULTS: Histopathologic study of four of the resected specimens confirmed the presence of CD, whereas in the fifth, there were features consistent with a remote encephaloclastic lesion. There were no postoperative deficits. Seizures in three of the patients were completely controlled at 2-3.5 years of follow-up; a fourth patient is still having a few seizures, which have required reinstitution of pharmacotherapy, and the fifth has obtained > or =70% control. All patients have had significant improvement in psychosocial measures. For comparison, five patients with generally similar clinical and neuroradiologic features to the previous group underwent preoperative ILTVE monitoring. The surgical outcomes between the two groups have not differed significantly. CONCLUSIONS: We conclude that patients with FL epilepsies may be able to undergo successful surgery without preoperative ILTVE monitoring, provided there is high concordance between neuroimaging tests (MRI, SPECT, PET) and the seizure phenotypes, even when routine EEGs and scalp LVTE fail to localize ictal onset unambiguously. The surgical outcomes of these patients generally paralleled those of the other subjects who also had FL epilepsy but who were operated on only after standard ILTVE monitoring.

Adolescent↗
PubMed1999

[The significance of morphological changes in the brain-tumor border area for the pathogenesis of brain edema in meningiomas: magnetic resonance tomographic and intraoperative studies].

PURPOSE: The aim of the study was to verify a possible correlation between macroscopic changes of the brain-tumor interface (BTI) and the development of a peritumoral brain edema in meningiomas. METHODS: 27 meningiomas were investigated in this prospective study using an optimized inversion-recovery (IR) sequence. After i.v. administration of 0.2 mmol Gd-DTPA/kg axial and coronary images were acquired (slice thickness = 2 mm). The distances of signal altered cortex and obliterations of the subarachnoid space (SAS) were measured at the BTI and related to the pial tumor circumference (cortical-index and SAS-index). Intraoperatively the BTI was divided into the following categories: 0: SAS not obliterated, 1: SAS partially obliterated, 2: direct contact between tumor and white matter, 3: tumor infiltration into brain. RESULTS: Edema-associated meningiomas showed a significantly (p = 0.0001) increased SAS-index (0.47 vs. 0.07) and cortical index (0.45 vs. 0.0) compared to cases without edema. Intraoperatively 95% of meningiomas with brain edema showed SAS-obliterations, compared to 50% of cases without an edema. CONCLUSIONS: Arachnoid adhesions at the BTI with obliteration of the SAS seem to play an essential role in the induction of brain edema in meningiomas.

Adult↗
PubMed1988

Identification and arrangement of the DNA sequence recognized in specific transformation of Neisseria gonorrhoeae.

DNA segments from Neisseria gonorrhoeae, cloned and propagated in Escherichia coli, were tested for the ability to competitively inhibit gonococcal transformation. The nucleotide sequences of active segments were determined and compared; these sequences contained the sequence 5' GCCGTCTGAA 3' in common. Subcloning studies confirmed the identity of this sequence as the gonococcal DNA recognition site. The three instances of the recognition sequence isolated from N. gonorrhoeae chromosomal DNA contain the sequence in the immediate neighborhood of its inverted repeat. Because a single copy of the sequence functions as a recognition site, the inverted duplication is not required for specific binding. The dyad symmetric arrangements of the chromosomal recognition sequences may form stable stem-loop structures that can function as terminators or attenuators of transcription. These inverted repeats are located at the boundaries of long open reading frames. The recognition sequence also constitutes part of two other probable terminators of gonococcal genes. We conclude that the signal for recognition of transforming DNA by gonococci is a frequent component of transcriptional terminator sequences. This regulatory function might account for the origin and maintenance of recognition sequences in the chromosomes of Gram-negative transformable bacteria.

Base Sequence↗
PubMed1992

The Saccharomyces cerevisiae ARG4 initiator of meiotic gene conversion and its associated double-strand DNA breaks can be inhibited by transcriptional interference.

In the yeast Saccharomyces cerevisiae, as in other eukaryotes, some regions of the genome have a much higher level of meiotic gene conversion than others. Previous deletion analysis indicated that the sequence necessary for the high level of gene conversion within the ARG4 region defined an initiation site located between positions -316 and -37 [relative to the first base pair (+1) of the ARG4 coding sequence] of the ARG4 promoter. To test whether this sequence is sufficient to promote gene conversion in a novel chromosomal context, we inverted on the chromosome various DNA fragments including the implicated region and the ARG4 coding sequence. Surprisingly, these inversions resulted in the loss of the normal recombination properties and double-strand-break formation associated with this process. By Northern analysis, we found that a transcript traverses the ARG4 initiation site in these inversion mutants but not in the wild type. When transcription through this region was prevented by a transcription terminator, the activity of the initiation site and the formation of double-strand breaks were restored. From these results and from complementary deletion analysis in the normal ARG4 orientation, we conclude that the activity of the ARG4 initiation site requires protection from transcriptional interference.

Chromosome Inversion↗
PubMed2003

Structural changes in RepA, a plasmid replication initiator, upon binding to origin DNA.

RepA protein is the DNA replication initiator of the Pseudomonas plasmid pPS10. RepA dimers bind to an inversely repeated operator sequence in repA promoter, thus repressing its own synthesis, whereas monomers bind to four directly repeated sequences (iterons) to initiate DNA replication. We had proposed previously that RepA is composed of two winged-helix (WH) domains, a structural unit also present in eukaryotic and archaeal initiators. To bind to the whole iteron sequence through both domains, RepA should couple monomerization to a conformational change in the N-terminal WH, which includes a leucine zipper-like sequence motif. We show for the first time that, by itself, binding to iteron DNA in vitro dissociates RepA dimers into monomers and alters RepA conformation, suggesting an allosteric effect. Furthermore, we also show that similar changes in RepA are promoted by mutations that substitute two Leu residues of the putative leucine zipper by Ala, destabilizing the hydrophobic core of the first WH. We propose that this mutant (RepA-2L2A) resembles a transient folding intermediate in the pathway leading to active monomers. These findings, together with the known activation of other Rep-type proteins by chaperones, are relevant to understand the molecular basis of plasmid DNA replication initiation.

Amino Acid Motifs↗
PubMed2004

Rapid evolution of beta-glucuronidase specificity by saturation mutagenesis of an active site loop.

Protein engineers have widely adopted directed evolution as a design algorithm, but practitioners have not come to a consensus about the best method to evolve protein molecular recognition. We previously used DNA shuffling to direct the evolution of Escherichia coli beta-glucuronidase (GUS) variants with increased beta-galactosidase activity. Epistatic (synergistic) mutations in amino acids 557, 566, and 568, which are part of an active site loop, were identified in that experiment (Matsumura, I., and Ellington, A. D. (2001) J. Mol. Biol. 305, 331-339). Here we show that site saturation mutagenesis of these residues, overexpression of the resulting library in E. coli, and high throughput screening led to the rapid evolution of clones exhibiting increased activity in reactions with p-nitrophenyl-beta-d-xylopyranoside (pNP-xyl). The xylosidase activities of the 14 fittest clones were 30-fold higher on average than that of the wild-type GUS. The 14 corresponding plasmids were pooled, amplified by long PCR, self-ligated with T4 DNA ligase, and transformed into E. coli. Thirteen clones exhibiting an average of 80-fold improvement in xylosidase activity were isolated in a second round of screening. One of the evolved proteins exhibited a approximately 200-fold improvement over the wild type in reactivity (k(cat)/K(m)) with pNP-xyl, with a 290,000-fold inversion of specificity. Sequence analysis of the 13 round 2 isolates suggested that all were products of intermolecular recombination events that occurred during whole plasmid PCR. Further rounds of evolution using DNA shuffling and staggered extension process (StEP) resulted in modest improvement. These results underscore the importance of epistatic interactions and demonstrate that they can be optimized through variations of the facile whole plasmid PCR technique.

Algorithms↗
PubMed2006

Retropharyngeal tendinitis: radiographic and magnetic resonance imaging findings.

PURPOSE: To describe the magnetic resonance imaging (MRI) findings in retropharyngeal tendinitis. MATERIAL AND METHODS: Within 1 year, four patients presenting with symptoms of retropharyngeal tendinitis were examined by radiography and MRI. RESULTS: On MRI and radiographs, all patients had characteristic soft-tissue swellings and calcifications related to the tendon of the longus colli muscle situated inferior to the anterior arc of C1. MRI showed well-defined edema, with high signal in the retropharyngeal tissue anterior to C1-C5 on short T1 inversion recovery (STIR) sequences, low signal on T1-weighted sequences, and low signal in the calcification on both sequences. In addition, three patients had high signal intensity changes on STIR sequences in the atlantoaxial joint situated posterior to the anterior arc of C1. CONCLUSION: MRI is a sensitive and accurate method in the diagnosis of retropharyngeal tendinitis. A new finding in this condition is an effusion or synovitis in the anterior atlantoaxial joint. MRI is a valuable tool in differentiating retropharyngeal tendinitis from other diagnoses such as retropharyngeal abscess, pyogenic spondylitis, and spondyloarthropathy.

Adult↗
PubMed2006

Magnetic resonance imaging and radiographic findings of seal finger.

PURPOSE: To describe the magnetic resonance imaging (MRI) and radiographic findings of five patients with seal finger. MATERIAL AND METHODS: The MR images and radiographs of five patients with seal finger were retrospectively evaluated. MRI was performed on four patients in the subacute phase, and follow-up imaging was done on one of them at 5 months. One patient had MRI only at a later stage 5 years after onset. Radiographs were taken three times in the subacute phase and once at a later stage. One patient had had seal finger in another finger previously. RESULTS: Short-tau inversion-recovery (STIR) sequence showed extensive subcutaneous soft tissue edema in all four patients in the subacute phase and tenosynovitis of the flexion tendons in two cases. Three patients had edema in 2-3 phalanges, and effusion in the distal interphalangeal (DIP) joint was seen in one case. At the later stage, no signal pathology in soft tissues or bones was seen in STIR images. In the subacute phase, radiographs showed digital soft-tissue swelling in three patients, and one patient had a narrowed DIP joint, periarticular osteoporosis, and a periosteal reaction. At the later stage, flexion contracture of the finger was seen. CONCLUSION: In addition to soft-tissue infection, seal finger causes bone marrow edema, tenosynovitis, and effusion in the interphalangeal joints visible as increased signal intensity in STIR images. Radiographs reveal periarticular osteoporosis with loss of cartilage in the subacute phase and flexion contracture at the later stage. MRI (STIR) allows more precise delineation of the inflammatory process compared to radiography.

Adult↗
PubMed1986

Longitudinal relaxation time measurements with non-uniform tilt angles.

The effect of non-uniform tilt angles across the slice thickness on the determination of T1 in NMR imaging is investigated in saturation-recovery, spin-echo and inversion-recovery pulse sequences. It is shown that for each of these imaging protocols, non-uniform tilt angles imply a multi-exponential dependence of NMR signal on the pulse repetition interval TR.

Magnetic Resonance Spectroscopy↗
PubMedDate not supplied

Genetic variability and mapping of peptidase-4 in Anopheles albimanus.

Field-collected and laboratory populations of Anopheles albimanus were analyzed for the presence of variability for the enzyme, peptidase. Four zones of peptidase electromorphs were observed. The Peptidase-4 (Pep-4) locus was analyzed genetically and assigned to a location adjacent to the centromere on the right arm of chromosome 2 by employing crosses involving morphological mutants, allozyme markers, a holandric translocation, and pericentric inversions. The gene sequence (and map distances) on chromosome 2 beginning from the left arm is: brown larva--40--ebony (eb)--centromere--pep-4--?--bent(be)--?--Glutamate oxaloacetate transaminase (Got)--11--Glucose oxidase-2(Go-2)--17--green larva (gl)--9--amber (am)--2--propoxur resistance (prr)--2--red eye (re)--21--6-Phosphogluconate dehydrogenase (6-Pgd)--1--yellow. The crossover frequencies between eb and Pep-4 and Pep-4 and Got were 34 and 16, respectively.

Alleles↗
PubMed1991

Gd-DTPA-enhanced short repetition time and short inversion time inversion recovery magnetic resonance imaging. Experimental and clinical assessment.

To suppress both water and fat signal while retaining the high signal of Gd-DTPA enhancement, magnetic resonance imaging (MRI) of phantoms and 28 patients with mass lesions was done using short repetition time (TR) and short inversion time inversion recovery (STIR) sequences. Optimal STIR pulse sequences of 500 to 1000/80-100/20-30 (TR/TI/TE) were determined by an experimental study. In most instances, a signal bandwidth of +/- 8 kHz was used to increase the signal-to-noise ratio. The authors measured image contrast between lesions and adjacent fatty tissue and compared postcontrast STIR and T1-weighted spin-echo (T1-W SE) images. When the signal intensity of a lesion is 80% of adjacent fatty tissue on postcontrast T1-W SE, short TR STIR images provide better tumor delineation.

Contrast Media↗
PubMed1983

Signal, noise, and contrast in nuclear magnetic resonance (NMR) imaging.

Calculations of the sensitivity of the saturation recovery and inversion recovery pulse sequences used in nuclear magnetic resonance imaging show the former to be superior in discriminating between tissues with the same proton density but different T1's. Two other pulse sequences, which are combinations of the above, have also been analyzed. These have lower T1 discrimination sensitivity, but other considerations, such as self-normalization, may still make them attractive. The calculations are only valid for selective excitation pulse sequences in which the selected slice profiles are approximately rectangular, and thus a sin(bt)/t radiofrequency excitation is desirable. In order to ensure that the saturation recovery sequence gives valid results for pulse repetition times comparable to or shorter than T2, it is necessary to destroy the coherence between pulse applications. For this purpose we use a series of "spoiler" gradient pulses between pulse trains. The saturation recovery pulse sequence also has the advantage that, by the correct choice of interpulse spacing, sensitivity close to the optimum T1 discrimination can be achieved over a wide range of T1 values. This has the potential advantage to the clinician of simplifying his choice of parameters for imaging.

Humans↗
PubMed1984

MR imaging of orbital tumors with CT and ultrasound correlations.

Fourteen patients with various orbital space-occupying lesions were studied by magnetic resonance (MR) imaging with a 0.15 T imager using spin-echo and inversion-recovery pulse sequences. Computed tomographic and ultrasonographic (US) examinations were done in 13 of these patients. All 14 lesions could be demonstrated by MR. Magnetic resonance was found to be better than US but not as good as CT in providing morphological details. Regarding tissue characterization, US was the most useful among the three modalities. With the T2-weighted spin-echo pulse sequence, a possible trend in tissue characterization could be shown. From our preliminary experience, we conclude that MR imaging using a 0.15 T resistive magnet offers no distinct advantage over the combination of CT and US in the diagnosis of orbital tumors.

Adult↗
PubMedDate not supplied

Iron-EHPG as an hepatobiliary MR contrast agent: initial imaging and biodistribution studies.

The paramagnetic metal complex iron(III) ethylenebis-(2-hydroxyphenylglycine) [Fe(EHPG)-] is an effective hepatobiliary contrast agent for liver enhancement in magnetic resonance (MR) imaging. The intravenous administration of 0.2 mmol/kg of Fe(EHPG)- to rats yields a 200% increase in the signal intensity of the liver when using a T1-weighted inversion recovery pulse sequence on a 1.4 T imaging system. Biodistribution studies in rats and a rabbit, along with imaging studies in a dog at 0.6 T, confirm that the complex has significant hepatocellular uptake and appears to be excreted unaltered into the bile. Control experiments with a different iron complex, iron(III) diethylenetriaminepentaacetic acid, reveal little hepatic affinity and poor enhancement capability due to its extracellular distribution. This initial evaluation of Fe(EHPG)- demonstrates that paramagnetic metal complexes with hepatobiliary specificity are well suited for enhancement of normal liver parenchyma and may increase the sensitivity of MR in the detection of liver disease.

Animals↗