Novel and recurrent TRPV4 mutations and their association with distinct phenotypes within the TRPV4 dysplasia family.

BACKGROUND: Mutations in TRPV4, a gene that encodes a Ca(2+) permeable non-selective cation channel, have recently been found in a spectrum of skeletal dysplasias that includes brachyolmia, spondylometaphyseal dysplasia, Kozlowski type (SMDK) and metatropic dysplasia (MD). Only a total of seven missense mutations were detected, however. The full spectrum of TRPV4 mutations and their phenotypes remained unclear. OBJECTIVES AND METHODS: To examine TRPV4 mutation spectrum and phenotype-genotype association, we searched for TRPV4 mutations by PCR-direct sequencing from genomic DNA in 22 MD and 20 SMDK probands. RESULTS: TRPV4 mutations were found in all but one MD subject. In total, 19 different heterozygous mutations were identified in 41 subjects; two were recurrent and 17 were novel. In MD, a recurrent P799L mutation was identified in nine subjects, as well as 10 novel mutations including F471del, the first deletion mutation of TRPV4. In SMDK, a recurrent R594H mutation was identified in 12 subjects and seven novel mutations. An association between the position of mutations and the disease phenotype was also observed. Thus, P799 in exon 15 is a hot codon for MD mutations, as four different amino acid substitutions have been observed at this codon; while R594 in exon 11 is a hotspot for SMDK mutations. CONCLUSION: The TRPV4 mutation spectrum in MD and SMDK, which showed genotype-phenotype correlation and potential functional significance of mutations that are non-randomly distributed over the gene, was presented in this study. The results would help diagnostic laboratories establish efficient screening strategies for genetic diagnosis of the TRPV4 dysplasia family diseases.

The DD genotype of angiotensin converting enzyme polymorphism is a risk factor for coronary artery disease and coronary stent restenosis in Japanese patients.

Stent implantation has decreased the incidence of restenosis after coronary intervention, but has not eliminated it. The contribution of the angiotensin-converting enzyme (ACE) genotype to the development of coronary artery disease and restenosis after coronary stenting was investigated in 67 Japanese patients in whom 103 lesions in which stents had been successfully implanted were assessed by quantitative coronary angiography, before, immediately after coronary stenting, and during follow-up. The distribution of the patients with the DD, ID, and II genotypes was 13%, 54%, and 33%, respectively. The prevalence of multivessel disease in the DD genotype was significantly higher (DD genotype: 78%; ID genotype: 58%; II genotype: 27%, chi2=8.13, p=0.016) and the late loss in the DD genotype (1.43+/-0.96 mm) was significantly greater (ID genotype: 0.78+/-0.98 mm and II genotype: 0.79+/-0.88 mm, p<0.05 vs DD genotype). However, there was no significant difference in the restenosis rate among the 3 genotypes. The present study in Japanese patients indicates that the DD genotype is associated with more extensive coronary artery disease and progression of the inward remodeling within the stented lesion, which is primarily caused by neointimal hyperplasia.

Phylogenetic analysis of the Erysipelothrix rhusiopathiae and Erysipelothrix tonsillarum based upon 16S rRNA.

The nucleotide sequences of 16S rRNA genes of Erysipelothrix rhusiopathine and Erysipelothrix tonsillarum were determined. The sequences are almost similar (99.8%) with only three nucleotides mismatched.

Detection and investigation of Campylobacter jejuni by polymerase chain reaction-restriction fragment length polymorphism analysis.

A molecular typing approach for Campylobacter jejuni with restriction fragment length polymorphism (RFLP) analysis of the flagellin gene flaA in C. jejuni, was generated and studied. Using polymerase chain reaction (PCR)-RFLP with the restriction endonuclease Mbo I, it was demonstrated that C. jejuni could be divided into four types. Genotypic analysis of C. jejuni by PCR-RFLP is a valuable technique for epidemiological typing.

Stature and severity in multiple epiphyseal dysplasia.

We investigated the stature and radiological findings in 15 patients with multiple epiphyseal dysplasia (MED). They were divided into normal-stature and short-stature groups according to their body height after 4 or 5 years of age. Their stature was not related to the involvement of the spine or epiphyses of long tubular bones except for the distal radius. Proximal phalanges and metacarpi were shorter in the short-stature group than in the normal-stature group, indicating that stature in MED had some relationship to the involvement of the wrist and hand. However, some patients in the normal-stature group showed involvement of distal radial epiphyses, and some patients in the short-stature group did not have stubby fingers. There are thus no clear-cut criteria to differentiate between the severe Fairbank type and the milder Ribbing type of MED.

Percutaneous transluminal coronary angioplasty for morphologic left anterior descending artery lesion in a patient with dextrocardia. A case report and literature review.

A case report of percutaneous transluminal coronary angioplasty (PTCA) to treat coronary atherosclerotic lesions in a patient with dextrocardia associated with situs inversus totalis is presented. The patient was a sixty-two-year-old man who was admitted with a diagnosis of congestive heart failure. Cardiac catheterization was performed. Left ventriculography showed mild hypokinesis in segments 2 and 3 with ejection fraction of 63%. Coronary arteriography revealed 74% stenosis in segment 7 of the left anterior descending (LAD) artery. PTCA for this lesion was performed. Successful dilation was achieved with the residual stenosis in the LAD reduced from 74% to 34%. Performance of PTCA in patients with dextrocardia is extremely rare, and only 8 cases have been reported to date. However, by visualizing the procedure as a mirror image and choosing a guide catheter that permits good engagement, it appears possible to perform it like ordinary PTCA.

Unusual right coronary artery runoff with right and left main coronary artery aneurysm. A case report.

A rare case of unusual right coronary artery runoff with right and left main coronary artery aneurysm is presented. The discussion is mainly from the etiologic point of view.

[Two cases of peripartum cardiomyopathy].

Two cases of peripartum cardiomyopathy, a rare type of dilated cardiomyopathy, are reported. A 36-year-old woman developed congestive heart failure 1 month after delivering her third child. Cardiac catheterization revealed diffuse hypokinesis of the left ventricle and an ejection fraction of 28%. The second study, 20 months later, demonstrated an ejection fraction of 46%. Endomyocardial biopsy showed mild interstitial edema. A 42-year-old woman developed toxemia during pregnancy. She delivered her second child at 38 weeks of gestation. Two weeks later, she developed congestive heart failure. Cardiac catheterization demonstrated diffuse hypokinesis of left ventricle with an ejection fraction of 40%. Endomyocardial biopsy revealed dense fibrosis. Follow-up angiography performed 8 months later showed similar findings with an ejection fraction of 34%. These two cases suggest the importance of evaluation of endomyocardial biopsy to determine the degree of interstitial fibrosis that may reflect the prognosis for patients with peripartum cardiomyopathy confirmed by measurements of ejection fraction.

Reaction of hydroxyapatite-sol in bone marrow.

Hydroxyapatite-sol was prepared by dispersing hydroxyapatite microcrystals into saline. The hydroxyapatite microcrystals were synthesized by neutralization reaction of calcium hydroxide suspension and phosphoric acid solution. Sizes of the hydroxyapatite microcrystals were less than 0.1 microns, and the mean value was 0.05 microm. The 0.2 ml hydroxyapatite-sol with a concentration of 14.8 mg/ml was injected into medullary cavities of rat femurs. Both sintered hydroxyapatite powder suspension and saline as comparative materials were injected into the medullary cavities in the same way. Reaction of the hydroxyapatite-sol in the bone medullary cavities was investigated histologically using light and transmission electron microscopes. After 3 days, new bone formation was observed by injection of hydroxyapatite-sol, while no bone formation was observed by injection of sintered hydroxyapatite powder and saline. Osteoblasts were observed endocytosing the hydroxyapatite-sol in the medullary cavities of the rats. Macrophages and undifferentiated osteoblasts were found around the hydroxyapatite-sol aggregation by transmission electron microscope. After 5 days, amounts of new bone increased and matured, forming trabeculae. Many osteoblasts were observed in a line along the surface of the bone. On the other hand, 5 days after injection of sintered hydroxyapatite powder and saline to bone formation was observed while at 10 days after injection, some immature new bone formation started to be observed. New bone increased and matured at 15 days postoperatively. From these results, it was concluded that hydroxyapatite-sol only quickly promotes the formation of new bone in bone marrow and can be used as injection liquid of new biomaterials for bone formation.

The behavior of pentaerythritol tetranicotinate in rat gastrointestinal tract as a prodrug.

The gas chromatographic assay method for pentaerythritol tetranicotinate, a nicotinic acid prodrug, and its hydrolysates was developed. The behavior of the drug in gastrointestinal tract was investigated in rat by using the method. The disappearance and hydrolysis of the drug were not observed in the gastric loop until 30 min. The rate of disappearance from the intestinal loop was 36.7% at 30 min which was significantly smaller than that of nicotinic acid. Little hydrolysis of the drug was observed in the buffer solution, pH 7.4, at 37 degrees up to 2 hr. However, the consecutive hydrolysis was observed when the drug was incubated with everted intestine or plasma. As to the rate of hydrolysis of the drug and its esterform hydrolysates by scraped intestinal mucosa, the ester to which the larger number of nicotinic acid was bound was hydrolyzed more rapidly. These results indicate that the orally administered drug is enzymatically hydrolyzed in the intestinal mucosa by a consecutive reaction. Although the hydrolysis rate of pentaerythritol tetranicotinate is rapid, the rate of its ester-form hydrolysate becomes slower gradually as the nicotinic acid is released. The released nicotinic acid is rapidly absorbed. The behavior of the drug revealed in this study suggests that pentaerythritol tetranicotinate is useful as a prodrug of nicotinic acid.

A highly topically active corticosteroid. Preliminary report.

The local anti-inflammatory effects of a synthetic corticosteroid, 17 alpha, 21-diacetory-2-bromo-6 beta, 9 alpha-difluoro-11 beta-hydroxy-1,4,-pregnadiene-3,20-dione (F-2), were evaluated by granuloma and granuloma pouch method. F-2 had an intensive and probably persistent topical anti-inflammatory activity. The systemic effects of F-2 were evaluated by measuring the thymus and adrenal gland weight. F-2 did not induce significant systemic effects at as high a dose as 125 times the effective dose.

Release characteristics of dibucaine dispersed in konjac gels.

A possible use of konjac gel for sustained release of drugs was examined in a monolithic system containing dibucaine. Dibucaine was dispersed in the gel which was prepared by gelation of the konjac flour in a borax solution at 60 degrees. The cumulative amount of the drug released plotted against the square root of time was linear in the monolithic system. This relationship was in agreement with that expected from the theoretical equation for planar configuration. The mechanism of the release of the drug from the gel may be considered to be leaching of the drug by the permeating fluid. The release profile from dried konjac gel was similar to that from undried gel, but that from unwarmed gel showed a deviation from linearity although sustained release was similarly obtained.