Influences of age and sex on the validity of bone scintigraphy for the diagnosis of temporomandibular joint osteoarthritis.

Bone scintigraphy has been used for the diagnosis of early-stage temporomandibular joint (TMJ) osteoarthritis (OA) owing to its high sensitivity. However, the diagnostic value of bone scintigraphy may be compromised when applied to patients in an age range with high bone metabolism rates. The aim of this study was to investigate the validity of bone scintigraphy as an appropriate diagnostic modality for TMJ OA. A total of 406 subjects (145 male, 261 female; age range 14-87 years) were selected, and all subjects underwent both bone scintigraphy and computed tomography (CT). The diagnosis of TMJ OA was confirmed with CT. Images obtained with bone scintigraphy were analyzed by visual and quantitative methods using the TMJ-to-skull ratio. The TMJ-to-skull ratio was significantly higher during adolescence and elderly adulthood, but differences between the sexes were not significant. The diagnostic value of the TMJ-to-skull ratio was lower in elderly adulthood in both males and females. The diagnostic utility of visual assessment was also compromised during late adulthood in both males and females. Thus bone scintigraphy has little value in the detection of TMJ OA, because the results could be influenced by age-related bone metabolism rates.

The influence of etiology and treatment factors on intellectual outcome in congenital hypothyroidism.

To determine the effects of hypothyroidism and hormonal patterns on outcome, we tested 65 7- to 12-year-old children with congenital hypothyroidism using standardized tests of intelligence, neuropsychological functioning, memory, and achievement. Results were analyzed by etiology, time to thyrotropin normalization, and hormone levels at testing. Children with athyreosis scored below other etiologies on visuospatial, attention, and arithmetic indices. Children whose thyroid-stimulating hormone levels normalized by 1 to 2 months of age scored higher than later normalizers on indices of visual memory, attention, and arithmetic. Normalization of thyroid-stimulating hormone by 3 months of age was associated with better memory and learning abilities than later normalization. Thyroid hormone levels at testing were correlated with indices of sensorimotor, spatial, and language abilities. Two children with persistently elevated thyrotropin levels were not adversely affected. Present findings signify the need to establish etiology, normalize thyrotropin early, and maintain hormone levels in the normal range throughout childhood in children with congenital hypothyroidism.

Analysis of expressed sequence tags from Brassica rapa L. ssp. pekinensis.

Non-redundant expressed sequence tags (ESTs) were generated from six different organs at various developmental stages of Chinese cabbage, Brassica rapa L. ssp. pekinensis. Of the 1,295 ESTs, 915 (71%) showed significantly high homology in nucleotide or deduced amino acid sequences with other sequences deposited in databases, while 380 did not show similarity to any sequences. Briefly, 598 ESTs matched with proteins of identified biological function, 177 with hypothetical proteins or non-annotated Arabidopsis genome sequences, and 140 with other ESTs. About 82% of the top-scored matching sequences were from Arabidopsis or Brassica, but overall 558 (43%) ESTs matched with Arabidopsis ESTs at the nucleotide sequence level. This observation strongly supports the idea that gene-expression profiles of Chinese cabbage differ from that of Arabidopsis, despite their genome structures being similar to each other. Moreover, sequence analyses of 21 Brassica ESTs revealed that their primary structure is different from those of corresponding annotated sequences of Arabidopsis genes. Our data suggest that direct prediction of Brassica gene expression pattern based on the information from Arabidopsis genome research has some limitations. Thus, information obtained from the Brassica EST study is useful not only for understanding of unique developmental processes of the plant, but also for the study of Arabidopsis genome structure.

NTR1 encodes a floral nectary-specific gene in Brassica campestris L. ssp. pekinensis.

We have characterized a gene specifically expressed in the floral nectaries of Brassica campestris L. ssp. pekinensis. Differential screening led to the isolation of a floral nectary-specific cDNA clone. Northern hybridization indicated that its mRNA transcript is 1450 nucleotides long and specific to the flower base. In situ hybridization and immunolocalization showed that its mRNA and protein are localized specifically to both the lateral and median nectaries of flowers. The cDNA codes for a 43.8 kDa polypeptide 392 amino acids long. The protein was named nectarin1 (NTR1) after floral nectary protein. NTR1 was located in the cytoplasm of nectariferous cells in the nectaries and was also observed in nuclei at a much lower level. The level of the transcript increases with flower development, especially during nectary development, but decreases abruptly with the opening of the flower. Genomic Southern blot analysis indicated that at least three copies of homologous genes were present in the genome of B. campestris, but that only a single copy was present in both Arabidopsis thaliana and Lycopersicon esculentum. The deduced amino acid sequence of NTR1 shows similarity to S-adenosyl-L-methionine:salicylic acid carboxyl methyltransferase of Clarkia breweri which is expressed mostly in petals. The function of the gene is speculated to be involved in the methylation of a plant secondary metabolite in the floral nectaries.

Satellite cereal yellow dwarf virus-RPV (satRPV) RNA requires a douXble hammerhead for self-cleavage and an alternative structure for replication.

The 110 nt hammerhead ribozyme in the satellite RNA of cereal yellow dwarf virus-RPV (satRPV RNA) folds into an alternative conformation that inhibits self-cleavage. This alternative structure comprises a pseudoknot with base-pairing between loop (L1) and a single-stranded bulge (L2a), which are located in hammerhead stems I and II, respectively. Mutations that disrupt this base-pairing, or otherwise cause the ribozyme to more closely resemble a canonical hammerhead, greatly increase self-cleavage. In a more natural multimeric sequence context containing the full-length satRPV RNA and two copies of the hammerhead, wild-type RNA cleaves much more efficiently than in the 110 nt context. Mutations in the upstream hammerhead, including a knock-out in the catalytic core, affect cleavage at the downstream cleavage site, indicating that multimers of satRPV RNA cleave via a double hammerhead. The double hammerhead includes base-pairing between two copies of the L1 sequence which extends stem I. Disruption of L1-L1 base-pairing slows cleavage of the multimer. L1-L2a base-pairing is required for efficient replication of satRPV RNA in oat protoplasts. Mutations that affect self-cleavage of the multimer do not correlate with replication efficiency, indicating that the ability to self-cleave is not a primary determinant of replication. We present a replication model in which multimeric satRPV RNA folds into alternative conformations that cannot form in the monomer. One potential metastable intermediate conformation involves L1-L2a base-pairing that may facilitate formation of the double hammerhead. However, we conclude that L1-L2a also performs some other essential function in the satRPV RNA replication cycle, because the L1-L2a base-pairing is more important than efficient self-cleavage for replication.

Molecular characterization of the garlic virus X genome.

The complete nucleotide sequence of the cDNA genome for garlic virus X (GVX), one of the major viruses infecting garlic plants, was determined. GVX is a single-stranded positive-sense RNA virus consisting of 8106 nucleotides excluding the 3'-end poly(A) tail and contains six open reading frames (ORFs) which encode putative proteins of 174 kDa (ORF1), 26 kDa (ORF2), 12 kDa (ORF3), 32 kDa (ORF4), 26 kDa (ORF5) and 15 kDa (ORF6). The putative viral proteins show similarity to those of carlaviruses and potexviruses but show the highest homology to shallot virus X (ShVX). Even though the GVX genome contains most of the structural elements common to carlaviruses and potexviruses, it is distinguished from them by the presence of an ORF4 which encodes an unusual protein. These results suggest that GVX may belong to an unassigned group of ShVX and GarV-type viruses rather than to the carlaviruses or potexviruses.

Identification of one of the major viruses infecting garlic plants, garlic virus X.

A partial cDNA clone for garlic virus X (GVX) was isolated. GVX was identified immunologically with an antibody raised against the recombinant coat protein (CP) and demonstrated to be one of the major viruses infecting garlic plants showing mosaic or streak symptoms. GVX belongs to an unassigned group of ShVX and GarV-type viruses rather than to carlaviruses or potexviruses. The recombinant CP of GVX was purified by Ni(2+)-NTA affinity chromatography. Anti-GVX CP antibody was raised against the purified recombinant CP. GVX particle is flexuous, rod-shaped, and about 750 nm long as determined by immunoelectron microscopy. The extent of infection by GVX of garlic plants was analyzed by Northern or immunoblot analyses of individual garlic plants cultivated in different regions. These results showed that almost all of the garlic plants tested from 40 different regions including America, China, Japan, and Korea are infected with GVX.

The loop B domain is physically separable from the loop A domain in the hairpin ribozyme.

In order to understand the catalysis mechanism of the hairpin ribozyme, mutant ribozymes were constructed. The distance between the loop A domain and the loop B domain was extended by inserting various lengths of nucleotide linkers at the hinge region in cis mutants, or the domains were separated physically in a trans mutant. All the mutant ribozymes, including the trans mutant, could cleave substrate RNA at the predicted site. A cis mutant with a single nucleotide insertion exhibited cleavage activity about twice as high as that of the wild-type (wt) ribozyme. The insertion of 2-5 nucleotides (nt) gradually reduced the activity to the level of the wt ribozyme. Insertion of a longer linker, up to 11 nt, resulted in the reduction of activity to one half of that of the wt ribozyme. The ribozyme with a single nucleotide insertion at the hinge region seems to form a more suitable conformation for catalysis by three-dimensional fold-back of the loop B to loop A containing the cleavage site. The trans mutant, in which the A and B domains were physically separated, maintained a significant level of activity, suggesting that both domains are necessary for catalysis, but separable. These results demonstrate that interaction between the A and B domains results in catalysis.