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The T-cell receptor beta chain-encoding gene repertoire of a New World primate species, the cotton-top tamarin.

The New World primate, the cotton-top tamarin (Saguinus oedipus), expresses major histocompatibility complex (MHC) class I molecules with limited diversity. The uniqueness of the cotton-top tamarin MHC class I loci may contribute to this species' unusual susceptibility to viral infections and high incidence of ulcerative colitis. As a prelude to examining the effect of this limited MHC class I diversity on the tamarin CD8(+) T-cell receptor (TCR) repertoire, we identified expressed tamarin TCR beta chain (TCRB) cDNAs by anchored and inverse polymerase chain reaction. Sequence alignments and phylogenetic comparisons with human and rhesus macaque sequences identified homologues of 21 human variable (V) gene families. Only single variable region genes were identified in each of these tamarin VB families, with the exception of the VB 5, 9, and 13 families which were comprised of two or three distinct members. The multiple genes within these three VB families do not appear to have separate human homologues, but rather aligned equally well to a single human gene from their respective VB families. These genes appear to have arisen, therefore, by duplication of certain VB genes in the tamarin ancestors following their divergence from the lineage leading to Old World primates and hominoids. Homologues of 12 of the 13 human joining (J) region genes were also identified in the tamarin. Comparison of the proportion of nonsynonymous (pN) and synonymous (pS) substitutions occurring per site within tamarin variable region genes demonstrated a reduction in pN in the framework regions compared with pN in the presumed MHC contact regions (CDR1 and CDR2). Taken together, these findings illustrate that the TCR beta chain-encoding genes of the cotton-top tamarin are similar in structure and degree of complexity compared with their Old World primate and human counterparts.

Amino Acid Sequence↗

High-resolution MR imaging of the proximal zone of the lunotriquetral ligament with a microscopy coil.

OBJECTIVE: To evaluate high-resolution MRI of the proximal zone of the lunotriquetral ligament (LTL) using a microscopy surface coil with a 1.5 T scanner. DESIGN AND SUBJECTS: The proximal zone of the LTL was reviewed in 90 subjects (23 asymptomatic normal volunteers and 67 patients with suspicion of triangular fibrocartilage complex injury) with high-resolution MRI using a 47-mm microscopy surface coil. High-resolution MR images were obtained with gradient recalled echo (GRE) T2*-weighted sequence and short tau inversion recovery imaging, with a 1- to 1.5-mm slice thickness, a 50-mm field of view, an imaging matrix of 140-224x512 using zero fill interpolation, and 3-4 excitations. As a qualitative analysis, the LTL was classified in shape and signal intensity. RESULTS: The triangle-shaped low-signal-intensity LTL was identified in 77 of 90 subjects (85.6%) on GRE images. The triangle was classified as regular (41.1%), broad-based (20.0%), narrow-based (6.7%), or asymmetrical (17.8%). The bar-shaped ligament was seen in one patient, and unclassified ligaments were seen in 12 patients. All volunteers showed triangle-shaped LTL. The MR signal intensity of the proximal zone in the LTL was characterized as homogeneously low intensity (type 1; 33.8%), linear intermediate or high signal intensity traversing the distal surface of the LTL (type 2; 45.5%), and linear intermediate or high intensity traversing both distal and proximal surfaces of LTL (type 3; 20.8%). CONCLUSION: The proximal zone of the LTL showed a broad spectrum of normal variations in shape and signal intensity on high-resolution MR images with a microscopy coil.

Adolescent↗

MR imaging of the normal appendix in children.

Our objective was to assess the ability of MR imaging in the detection of the normal appendix, and to describe the MR appearance of the normal appendix. There were 15 healthy volunteers (11 girls, 4 boys; mean age 12.3 years) who underwent MR imaging on a 1.0-T unit. The imaging protocol included axial and coronal T2-weighted ultra turbo spin-echo (UTSE)-weighted images, axial T1-weighted turbo spin-echo (TSE) and coronal short tau inversion recovery (STIR)/TSE sequences. Confidence regarding the detection was scored from 1 (high confidence) to 3 (low confidence). Thickness was measured and MR appearance described. Clinical control after 2 weeks revealed no signs or symptoms of acute appendicitis. The normal appendix was seen in 86% on T2/UTSE-weighted images and in 73% on T1/TSE-weighted images and in none on STIR/TSE images. On axial T2/UTSE-weighted images, normal appendix had a hyperintense center and a hypointense wall, and was mostly hypointense on T1/TSE-weighted images, with a mean thickness of 4.5 mm. Magnetic resonance imaging seems to be an accurate method for the assessment of the normal appendix in children; thus, MR imaging might be an alternative to CT if US examinations are inconclusive.

Appendix↗

Scarred myocardium imposes additional burden on remote viable myocardium despite a reduction in the extent of area with late contrast MR enhancement.

Magnetic resonance imaging (MRI) can simultaneously detect and quantify myocardial dysfunction and shrinkage in contrast-enhanced areas postinfarction. This ability permits the investigation of our hypothesis that transformation of infracted myocardium to scarred tissue imposes additional burdens on peri-infarcted and remote myocardium. Pigs (n = 8) were subjected to reperfused infarction. Gd-DOTA-enhanced inversion recovery gradient echo sequence (IR-GRE) imaging was performed 3 days and 8 weeks postinfarction. Global and regional left ventricular (LV) function was evaluated by cine MRI. Triphenyltetrazolium chloride (TTC) stain was used to delineate infarction while hematoxylin and eosin (H & E) and Masson's trichrome stains were used to characterize remodeled myocardium. Late contrast-enhanced MRIs showed a decrease in the extent of enhanced areas from 17 +/- 2% at 3 days to13 +/- 1% LV mass at 8 weeks. TTC infarction size was 12 +/- 1% LV mass. Cine MRIs showed expansion in dysfunctional area due to unfavorable remodeling, ischemia, or strain. Ejection fraction was reduced in association with increased end-diastolic and end-systolic volumes. Scarred myocardium contained collagen fibers and remodeled thick-walled vessels embedded in collagen. Sequential MRI showed greater LV dysfunction despite the shrinkage in extent of enhanced areas 2 months postinfarction. The integration of late enhancement and cine MRI incorporates anatomical and functional evaluation of remodeled hearts.

Analysis of Variance↗

Comparison of a conventional cardiac-triggered dual spin-echo and a fast STIR sequence in detection of spinal cord lesions in multiple sclerosis.

The current optimal imaging protocol in spinal cord MR imaging in patients with multiple sclerosis includes a long TR conventional spin-echo (CSE) sequence, requiring long acquisition times. Using short tau inversion recovery fast spin-echo (fast STIR) sequences both acquisition time can be shortened and sensitivity in the detection of multiple sclerosis (MS) abnormalities can be increased. This study compares both sequences for the potential to detect both focal and diffuse spinal abnormalities. Spinal cords of 5 volunteers and 20 MS patients were studied at 1.0 T. Magnetic resonance imaging included cardiac-gated sagittal dual-echo CSE and a cardiac-gated fast STIR sequence. Images were scored regarding number, size, and location of focal lesions, diffuse abnormalities and presence/hindrance of artifacts by two experienced radiologists. Examinations were scored as being definitely normal, indeterminate, or definitely abnormal. Interobserver agreement regarding focal lesions was higher for CSE (kappa = 0.67) than for fast STIR (kappa = 0.57) but did not differ significantly. Of all focal lesions scored in consensus, 47% were scored on both sequences, 31% were only detected by fast STIR, and 22% only by dual-echo CSE (n.s.). Interobserver agreement for diffuse abnormalities was lower with fast STIR (kappa = 0.48) than dual-echo CSE (kappa = 0.65; n. s.). After consensus, fast STIR showed in 10 patients diffuse abnormalities and dual-echo CSE in 3. After consensus, in 19 of 20 patients dual-echo CSE scans were considered as definitely abnormal compared with 17 for fast STIR. The fast STIR sequence is a useful adjunct to dual-echo CSE in detecting focal abnormalities and is helpful in detecting diffuse MS abnormalities in the spinal cord. Due to the frequent occurrence of artifacts and the lower observer concordance, fast STIR cannot be used alone.

Analysis of Variance↗

Measurements of T1 and T2 relaxation times of colon cancer metastases in rat liver at 7 T.

The purpose of this study was to investigate the magnetic resonance imaging (MRI) characteristics of colon cancer metastases in rat liver at 7 T. A dedicated RF microstrip coil of novel design was built in order to increase the signal-to-noise ratio and, in combination with respiratory triggering, to minimize motion artifacts. T1- and T2-weighted MR imaging was performed to follow tumor growth. T1-weighted images provided a good anatomical delineation of the liver structure, while the best contrast between metastases and normal liver tissue was achieved with T2-weighted images. Measurements of T1 and T2 relaxation times were performed with inversion recovery FLASH and Carr-Purcell-Meiboom-Gill and inversion recovery FLASH imaging sequences, respectively, for quantitative MR characterization of metastases. Both the T1 and T2 of the metastases were significantly higher than those of normal liver tissue. Further, an increase in the T1 relaxation time of the metastases was observed with tumor growth. These findings suggest that quantitative in vivo MR characterization provides information on tumor development and possibly response to therapy, though additional studies are needed to elucidate the correlation between the changes in relaxation times and tumor microenvironment.

Animals↗

Homonuclear chemical shift correlation in rotating solids via RNnu n symmetry-based adiabatic RF pulse schemes.

The efficacy of RN(nu) (n) symmetry-based adiabatic Zero-Quantum (ZQ) dipolar recoupling schemes for obtaining chemical shift correlation data at moderate magic angle spinning frequencies has been evaluated. RN(nu) (n) sequences generally employ basic inversion elements that correspond to a net 180 degrees rotation about the rotating frame x-axis. It is shown here via numerical simulations and experimental measurements that it is also possible to achieve efficient ZQ dipolar recoupling via RN(nu) (n) schemes employing adiabatic pulses. Such an approach was successfully used for obtaining (13)C chemical shift correlation spectra of a uniformly labelled sample of (CUG)(97) - a triplet repeat expansion RNA that has been implicated in the neuromuscular disease myotonic dystrophy. An analysis of the (13)C sugar carbon chemical shifts suggests, in agreement with our recent (15)N MAS-NMR studies, that this RNA adopts an Alpha-helical conformation.

Nuclear Magnetic Resonance, Biomolecular↗

Sensitivity of renal brush-border Na+-cotransport systems to anions.

The effect of anions on Na+-cotransport of succinate, lactate, glucose, and phenylalanine was studied under voltage clamped conditions in brush-border membrane vesicles prepared from rabbit renal cortex. The initial rate of succinate uptake varied by an order of magnitude depending on the anion: the highest rates were obtained with fluoride and gluconate, and the lowest with iodide. The anion sequence corresponded with the inverse of the anion hydration energies. The kinetics of succinate uptake were measured in the presence of fluoride and chloride. There was no difference in the maximal rates of uptake, but the Kt in fluoride (0.30 mM) was less than half that in chloride (0.70 mM), i.e. Cl- behaved as a competitive inhibitor of succinate transport with a Ki of 150 mM. The uptake of L-lactate, D-glucose and L-phenylalanine was less sensitive to anions, and there was no correlation with hydration energies. We conclude that the anion effects on sugar and amino acid uptakes measured under open-circuit conditions are largely due to variations in membrane potential, but in the case of the dicarboxylate transporter anions behave as weak competitive inhibitors. The specificity of the anion inhibition suggests that the dicarboxylate binding sites have a weak field strength relative to water.

Animals↗

Measurement of lithium transport in RBC from psychiatric patients receiving lithium carbonate and normal individuals by 7Li NMR spectroscopy.

A reproducible 7Li nuclear magnetic resonance (NMR) method, based on a modified inversion recovery (MIR) pulse sequence, was used to discriminate between intra- and extracellular lithium concentrations in red blood cell (RBC) suspensions. The rates of Na(+)-Li+ countertransport determined by the 7Li NMR method were significantly correlated with the measurements made by atomic absorption (AA) for 14 psychiatric patients receiving lithium carbonate (r = 0.937) and 14 normal individuals (r = 0.931). As expected, the rates of Na(+)-Li+ countertransport measured by MIR were significantly lower for the psychiatric patients receiving lithium carbonate than for normal individuals. The 7Li NMR method provides RBC Li+ countertransport information comparable to AA for psychiatric patients and normal individuals. A description of the advantages of the 7Li NMR method in contrast to the AA method, including the study of Li+ interactions with RBC components such as membrane proteins and anionic phospholipids, is included.

Adult↗

Formalin fixed brains are useful for magnetic resonance imaging (MRI) study.

We carried out magnetic resonance imaging (MRI) studies on human brains which had been fixed in formalin solution for over 2 years and had been proven neuropathologically to be cases of multiple sclerosis (MS), progressive multifocal leukoencephalopathy (PML), and Balo's concentric sclerosis (Balo). Using spin echo (SE) and inversion recovery (IR) pulse sequences to detect demyelinated lesions in a living person with MS, the demyelinated lesions of the fixed brains in cases of MS, PML and Balo definitely re-appeared, although T1 and T2 in the gray and white matter were reduced following fixation. High signal areas on the SE images corresponded not only to the characteristic distribution of demyelinated lesions in the white matter but also to sparse myelin, gliosis and mild perivascular cuffing in the white matter around the demyelinated foci in cases of the fixed MS, PML and Balo brains. On the IR images, only MS plaques were evident. This MRI study of fixed brains proved useful to elucidate clinicopathological correlations.

Adolescent↗

Comparative sequence analysis of the inverted terminal repetition in the genomes of animal and avian adenoviruses.

The nucleotide sequences at the inverted terminal repetitions from two animal adenoviruses, infectious canine hepatitis virus and equine adenovirus, and from one avian adenovirus, CELO, were analyzed. DNAs from infectious canine hepatitis virus and equine adenovirus contain a homologous region which is 23 nucleotides long from the terminus. The first 17 nucleotides of this region are identical to the ones in human adenovirus type 2 DNA. The striking homologous sequence of 14 nucleotides, conserved in the inverted terminal repetitions of several human adenovirus strains and in simian adenovirus type 7, is only partially conserved in the two animal and one avian adenoviruses reported here.

Adenoviridae↗

Alternative splicing caused by RNA secondary structure.

mRNA precursors with stable hairpins were constructed by inserting inverted repeats into an adenovirus transcriptional template that encoded the three late leader exons. When the loop of the hairpin contained the second exon and the flanking splice sites, most of the RNA spliced in vitro had the first exon joined directly to the third exon. The remainder was spliced normally. The same types of alternatively spliced RNAs were formed when a similar template was introduced into HeLa cells by transfection. Thus both in extracts and in cells, an exon became optional when sequestered in a hairpin loop. Perhaps a related mechanism creates the alternative splicing patterns of complex transcription units.

Adenoviruses, Human↗

Identification by Tn10 transposon mutagenesis of host factors involved in the biosynthesis of K99 fimbriae of Escherichia coli: effect of LPS core mutations.

Tn10 transposon mutagenesis of Escherichia coli producing K99 fimbriae was carried out to identify host factors involved in regulation of biosynthesis of fimbriae. Two chromosomal mutants were obtained that showed a strongly reduced cell surface expression of K99 fimbriae upon colony blotting and ELISA. Analysis by inversed PCR and nucleotide sequencing showed that one mutant (EP14) contained the Tn10 transposon in rfaQ, affecting the expression of the rfaQGP gene cluster, whereas the other mutant (EP35) was affected in a, to date, unknown region of the genome. Immunoblotting analysis confirmed a Rd1 type of LPS of mutant strain EP14. These findings for the first time indicated an effect of LPS core biosynthesis on the biogenesis of fimbriae at the cell surface. Preliminary experiments indicated that K99 major subunits, in contrast to K88 subunits, strongly bind LPS molecules.

Antibodies, Monoclonal↗

The 3' and 5'-terminal sequences of influenza A, B and C virus RNA segments are highly conserved and show partial inverted complementarity.

The 3'- and 5'-terminal nucleotides of the genome segments of an influenza A, B, and C virus were identified by directly sequencing viral RNA using two different sequencing techniques. A high degree of conservation at the 3' ends as well as at the 5' ends was observed among the genome segments of each virus and among the segments of the three different virus types. A uridine-rich region was observed from positions 17 through 22 at the 5' end of each segment. Moreover, the conserved 3' and 5'-terminal sequences showed partial and inverted complementarity. This feature results in very similar sequences at the 3' ends of the plus and minus strand RNAs and may also enable single-strand RNAs of influenza virus to form "panhandle" structures. Inverted complementary repeats may play an important role in initiation of viral RNA replication.

Base Sequence↗

The plasmid cloning vector pBR325 contains a 482 base-pair-long inverted duplication.

The plasmid pBR325 is a cloning vector constructed in vitro by addition of the chloramphenicol resistance (Cmr) gene of an IS1-flanked transposon to pBR322 (Bolivar, 1978). It is a 5 995 bp plasmid carrying no sequence originating from IS1. DNA-sequence data suggest that its Cmr segment was derived from a Cm transposon longer than Tn9. The plasmid pBR325 carries between the Cmr and Tcr genes a 482 bp sequence which duplicates, in the opposite orientation, a section pf pBR322 located at the end of the tcr gene. The same structure was found in pBR328, a deletion derivative of pBR325 (Soberon et al., 1980). The possible implications of this inverted duplication on cloning experiments are discussed.

Base Sequence↗

Tissue contrast enhancement: image reconstruction algorithm and selection of TI in inversion recovery MRI.

It is clearly demonstrated that the proper application of the inversion recovery imaging pulse sequence is dependent on the method of image reconstruction and the selection of TI for optimum tissue contrast. There are two methods of 2DFT image reconstruction of IR sequence time-domain raw data. The first is a modulus-image reconstruction algorithm (contrast-obliterating option), and the second is a phase-correction routine for reconstructing "phase-sensitive" true IR-images. The second option generates proper "in-phase" images, retains proper scale of contrast, but can invert the algebraic sign of image-values under certain conditions. A series of "phase-sensitive" and "modulus" reconstructed brain images, obtained with conventional and optimized new IR pulse sequences, are shown to demonstrate these effects. They illustrate the considerable advantages gained, in practical clinical situations, if one generates "phase-sensitive" true IR-images from IR-sequence raw data at optimum TI for tissue contrast enhancement.

Algorithms↗

Methodology for the measurement and analysis of relaxation times in proton imaging.

Measurements of proton T1 and T2 were performed on GdCl3 solutions (20 less than T2 less than 500 msec, 90 less than T1 less than 1000 msec) on large-bore NMR imaging systems operating at 1.0T and 1.5T. CPMG multi-echo (ME), multiple saturation recovery (MSR) and modified fast inversion recovery (MFIR) pulse sequences as well as a sequence that combines and interleaves T1 and T2 weighted data acquisition (which we call "multiple saturation-recovery multiple-echo" (MSRME) were used. The relaxation data are compared to those obtained on a small bore NMR spectrometer operated at 1.5T. T1 and T2 values for the solutions were found to be the same within 10% for the two fields. Reproducibility of measurements of T1, T2 and the unnormalized spin density of the solutions was better than 5%. Systematic errors, amenable to correction through calibration, are noted in the imager T1 and T2 values. T1 and T2 values for some typical neural tissues at 1.5T and body tissue at 1.0T for human volunteers were obtained and are tabulated.

Gadolinium↗