Indometacin up-regulates TFF2 expression in gastric epithelial cells.

BACKGROUND: Trefoil factor family peptides are expressed in gastrointestinal epithelial cells and play a critical role in maintaining mucosal integrity. Although non-steroidal anti-inflammatory drugs (NSAIDs) are important causative agents of gastric mucosal lesions, few data are available about the effect of NSAIDs on trefoil family peptides in gastric mucosa. AIM: To examine whether indometacin, a widely used NSAID, affects trefoil factor family expression in gastric epithelial cells. METHODS: MKN45, a cell line derived from human gastric cancer, was used. TFF1, TFF2, and TFF3 mRNA expression was assessed by real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR). TFF2 gene transcription was also examined by luciferase reporter gene assay. RESULTS: Relative expression level of TFF1, TFF2, TFF3 mRNA was 616: 12: 1 in unstimulated MKN45 cells. Although indometacin (1-250 micro mol/L) had no significant effect on the expression of TFF1 and TFF3 mRNA, it up-regulated TFF2 mRNA expression in a dose- and time-dependent manner. Luciferase reporter gene assay confirmed the up-regulation of TFF2 gene transcription by indometacin. Indometacin-induced up-regulation of TFF2 expression was not antagonized by externally applied prostaglandin E2. CONCLUSION: These results suggest that indometacin up-regulates gastric epithelial cell TFF2 expression through a COX-independent mechanism. Since TFF peptides play an important role in gastric mucosal protection, indometacin-induced TFF2 may reduce the degree of gastric mucosal damage induced by indometacin.

Polymorphism of the polyalanine tract of thyroid transcription factor-2 gene in patients with thyroid dysgenesis.

OBJECTIVE: One of the thyroid-specific transcription factors, thyroid transcription factor-2 (TTF-2), performs a crucial role in the development of the thyroid gland. We performed genetic analysis of the TITF2 gene (encoding TTF-2) in patients with thyroid dysgenesis. METHODS: By direct sequencing of the PCR products of TITF2, we screened the genomic DNA from 46 patients with thyroid dysgenesis (five had agenesis, six had hypoplasia, 15 had ectopy, and 20 were undetermined). We also studied the transcriptional activities of TITF2 by co-expressing the luciferase gene directed by the human thyroglobulin gene promoter. RESULTS: Human TITF2 consists of a forkhead domain, a polyalanine tract, and unique C-terminal residues. In one of the patients with an ectopic sublingual thyroid, we found a polyalanine tract of 11 alanine residues on one chromosome instead of the 14 alanine residues found in normal controls. In one patient with hypoplasia, the polyalanine tract consisted of 12 heterozygous alanine residues. The reduced polyalanine tracts were not detected in 101 normal individuals. However, the expression study showed that the transcriptional activities of TITF2 with reduced polyalanine-tract lengths were equal to that of TITF2 with an unreduced polyalanine tract. CONCLUSION: These results suggest that the polymorphism of the polyalanine tract of TITF2 is not a frequent cause of developmental defects of the human thyroid gland.

A novel missense mutation (G2320R) in thyroglobulin causes hypothyroidism in rdw rats.

The rdw rat is a hereditary hypothyroid variant initially derived from the Wistar-Imamichi strain. Proteome analysis by two-dimensional gelelectrophoresis showed that molecular chaperones accumulated in the thyroid glands, suggesting retention of abnormal proteins in the endoplasmic reticulum (ER). Anatomical studies indicated that thyroglobulin (Tg) was not secreted into the follicular lumina, but retained in the dilated ER. Sequencing of the entire Tg complementary DNA from the rdw rat revealed a missense mutation (G2320R) in the acetylcholinesterase-like domain at the 2320th amino acid residue. Carbohydrate residues of the G2320R Tg mutant were of the high-mannose ER type, as shown by sensitivity to the treatment with endoglycosidase H. Molecular chaperones, GRP94, GRP78, and calreticulin, were all accumulated in the rdw rat thyroid glands. Computer analysis of protein secondary structure predicted that the mutation would cause extension of the helix where beta-sheet and turns were formed in the normal Tg. Altered folding of Tg might account for the impaired intracellular transport of Tg and activated premature degradation by the same mechanism as in ER storage diseases.

[Acute right ventricular infarction: assessment with radionuclide ventriculography].

The clinical significance of right ventricular (RV) infarction has been neglected compared with left ventricular infarction. In recent years, however, the clinical importance of RV function in the treatment of myocardial infarction has been well recognized. We performed prospective radionuclide studies to assess the incidence and prognosis of RV infarction in 50 cases of initial acute myocardial infarction (25 cases of anterior and 25 of inferior infarction). Radionuclide ventriculography was performed within the first two days after onset of symptoms, and repeated one-two weeks and one month after the attack, respectively. RV infarction was diagnosed by the presence of severe RV regional wall motion abnormalities and positive signs of at least one of the following diagnostic signs: ST elevation at V4R in the ECG, positive 99m-technetium pyrophosphate myocardial scintigram at the RV free wall, and positive right heart catheterization findings. Results were as follows: 1. RV infarction was documented in 15 of 25 cases with inferior infarction, but there were no cases in anterior infarction. 2. There were no remarkable changes of RV ejection fraction (EF) in anterior myocardial infarction during one month (41% +/- 8% in acute phase and 43 +/- 8% in four weeks later). However, RVEF was markedly improved from 34 +/- 11% during first two days, to 38 +/- 7% during one-two weeks, and 39 +/- 8% four weeks after the attack, in cases of inferior infarction with RV infarction. Without RV infarction, RVEF in cases of inferior infarction did not show improvement. 3. In 11 of 15 cases with RV infarction, RV regional wall motion abnormalities improved to the normal range, which seemed to contribute to the improvement of RVEF. 4. Hemodynamic findings with Swan-Ganz catheters showed typical findings compatible with RV infarction only in seven of 13 cases with RV infarction. Thus this finding implies that RV failure did not always accompany RV infarction. 5. Coronary arteriography revealed that right coronary arterial lesions proximal to the RV branch were documented in all 10 cases with RV infarction who had coronary arteriography. RV infarction, caused by deranged coronary blood flow at the RV branch of the right coronary artery, showed marked improvement of RVEF during four weeks after the attack in prospective radionuclide studies. This finding was not seen in left ventricular infarction. The pathophysiological mechanism of improvement of RVEF in RV infarction would be the difference of its coronary circulation compared with that of the left ventricle.(ABSTRACT TRUNCATED AT 400 WORDS)