BACKGROUND: Tyrosine phosphorylation of proteins has been a focus of extensive studies since it plays crucial roles in regulation of diverse biological reactions. To understand the involvement of tyrosine phosphorylation in kidney functions, a comprehensive proteomic study for tyrosine-phosphorylated proteins was performed in the normal rat kidney. METHODS: Two-dimensional gel electrophoresis and immunoprecipitation using anti-phosphotyrosine antibodies were employed to detect tyrosine-phosphorylated proteins. The proteins were analysed by mass spectrometry and validated by immunological analyses using specific antibodies. RESULTS: Most of tyrosine-phosphorylated proteins were confined to the glomerulus and predominantly localized along the glomerular capillary wall, especially in the foot processes of podocytes. Our systematic proteomic analysis identified nephrin, SHPS-1 (tyrosine-protein phosphatase non-receptor-type substrate 1), FAK1 and paxillin as major tyrosine-phosphorylated proteins and Neph1, talin and vinculin as minor tyrosine-phosphorylated proteins. In the present study, SHPS-1 was identified as a novel tyrosine-phosphorylated protein in the glomerulus and was also predominantly localized at the foot processes. Mass spectrometric analysis identified in vivo phosphorylation sites of SHPS-1 on Y460, Y477 and Y501. CONCLUSION: This study identified tyrosine-phosphorylated proteins in normal rat kidney, which were prominently rich in the glomerulus and localized at the podocyte foot processes. These proteins were categorized as cell-to-cell or cell-to-matrix adhesion complex-related molecules, suggesting their pivotal roles in the glomerular ultrafiltration.

In order to investigate the effects of dietary supplementation rich in omega 3 and omega 6 fatty acids, we set up an experiment of twenty four C57BL/6J male mice segregated into 3 groups: normal diet (ND), omega 3 polyunsaturated fatty acid (n-3 PUFA,) and omega 6 (n-6 PUFA). At the end of the experiment that lasted for 1 month, food consumption of ND and n-3 PUFA were similar while it decreased in n-6 PUFA group. Total cholesterol, triglycerides, free fatty acids, and phospholipids profiles were increased in n-6 PUFA. LDL decreased in n-3 PUFA while increased in n-6 PUFA fed mice comparing to control group. On the other hand, there was no difference between treatments in HDL and glucose levels. Expression of leptin (ob) gene transcripts in epididymal fat were significantly elevated in n-6 PUFA mice compared to ND and n-3 PUFA groups while hypothalamic ob receptor A (obRa) mRNA did not changed in response to diet regimes. Transmission and scanning electron microscopy showed different degrees in fatty changes in the liver of both PUFA groups including lipid droplet infiltration and Ito cells with over accumulated lipids. In conclusion, under PUFA dietary supplementation, the hyperlipidemic status and elevated ob expression of n-6 PUFA but not n-3 PUFA fed mice suggests altered lipid metabolism between PUFA groups and/or different endocrine involvement. Moreover, the coincidently structural changes observed in liver of this group direct us to call for further studies to investigate the anti-obesity effect and safety of these PUFA under high supplementation condition.

The efficiency of a porcine embryo vitrification method that uses water-soluble films of pullulan, a naturally-occurring polysaccharide polymer, was compared with two other types of vitrification methods using different devices and solutions for vitrification and warming. Blastocysts collected in vivo and vitrified by the conventional straw (ST), Cryotop((R))(MVC) or pullulan film vitrification (PFV) methods were stored in liquid nitrogen for a certain period of time, after which the cryoprotective agents were removed by stepwise dilution. Fresh embryos were used as controls for the non-vitrification group. The vitrified-warmed embryos were incubated in TCM199 with 0.1 mMbeta-mercaptoethanol and 20% fetal bovine serum for 24 h at 38.5 C in humidified air with 5% CO(2) to evaluate their viability. The survival rate of embryos in the ST group (48.3%) was significantly lower than that of those in the MVC (70.7%), PFV (79.0%) and non-vitrification (94.4%) groups. The oxygen consumption rate after vitrification was significantly lower than that before vitrification in the ST group, but was not significantly different in the MVC and PFV groups. Both the oxygen consumption rates of embryos after warming and the live cell numbers in the ST group were lower than those in the MVC group, while they did not differ significantly between the PFV and MVC groups. There was a correlation between the oxygen consumption rate and the number of live cells in vitrified embryos after warming. Our results demonstrated that in vivo-derived porcine embryos could be vitrified using pullulan films.

A 76-year-old woman had a chest pain and high fever, who was admitted to the intensive care unit diagnosed as acute pericarditis. Enhanced CT-scan showed a 47-mm aneurysm in the aortic arch which seemed to be impending rupture and the part of the aorta looked like a pseudoaneurysm. Emergent total aortic arch replacement with a rifanpicin-bonded Dacron graft was performed. Pericardial effusion was purulent and the aorta was infected with pus discharge in the aortic wall. There were some ulcerations on the surface of the luminal wall of the aorta. One of them was penetrating into the pericardial space causing a pseudoaneurysm. Both pericardial effusion and excised aortic wall were sent to culture study and resulted in positive for Streptococcus pneumonia. The infection of the aorta, with erosion into the pericardial space, seemed to be the cause of purulent pericarditis. Antibiotic therapy was commenced immediately after surgery and continued for four weeks. Though she had neurological deficit after surgery, her infection was well controlled and there was no recurrence of infection eleven months after surgery. Keywords: Infected aortic aneurysm; Aortic arch; Streptococcus pneumonia; Purulent pericarditis.

Purpose: There is little knowledge regarding the prevalence of mucosal injury (MI) in Japanese patients receiving antiplatelet therapy. This study estimated the prevalence of gastroduodenal MI in asymptomatic Japanese patients taking antiplatelet agents and nonsteroidal anti-inflammatory drugs (NSAIDs). Methods: Among patients who underwent an upper gastrointestinal endoscopy at Teikyo University Hospital (Tokyo, Japan), 382 asymptomatic patients taking either low-dose aspirin, ticlopidine, cilostazol, or other NSAIDs and 119 people not taking any of these agents were included. Endoscopic records were evaluated for the presence of MI. Results: Aspirin and NSAIDs users showed a higher prevalence of MI than controls (Aspirin, OR = 2.6 (95% CI = 1.4 - 4.9), NSAIDs, 2.9 (1.4 - 4.4)). Concomitant use of aspirin and NSAIDs increased the prevalence of MI (11.2 (2.8 - 44.8)). Ticlopidine and cilostazol were less likely to cause injury than aspirin and other NSAIDs, the difference remained insignificant due to small sample number (ticlopidine, 0.8 (0.2 - 4.0), cilostazol, 1.3 (0.3 - 4.8)). Conclusions: In asymptomatic Japanese patients receiving low-dose aspirin, the prevalence of the gastroduodenal MI was the same as that in patients taking NSAIDs and higher than that in controls.

The relationship between bronchovascular cuff formation and lung lymph flow in hydrostatic edema was evaluated. After a balloon was inserted into the left atrium to increase left atrial pressure for 5 hrs, peripheral lung tissues were resected for analysis of the wet-dry ratio and cuff formation. The degree of cuff formation was expressed as the cuff ratio (outer diameter of cuff/outer diameter of microvessel or airway) in three size categories: 80-200, 200-400, and 400-750 microm in diameter. The amount of excess lung lymph (Ex LL) for 5 hrs was calculated from the recorded data for the whole lymph flow wave. The wet-dry ratio showed a significant correlation with ALAP and lung lymph flow increased significantly (flow rate, 0.67 +/- 0.46 ml/min (mean +/- SD); Ex LL, 56.4 +/- 47.6 ml). Cuff formation was found at all levels of the bronchovascular tree, with a larger cuff ratio (> 1.3) observed at arteries and veins of 80-200 microm in diameter, but a significant correlation with Ex LL was found only for arteries of 80-200 microm. Fluid accumulation in lung interstitium first occurred at smaller extra-alveolar arteries even under mild hydrostatic pressure elevation with a significant increase in lymph flow.

PURPOSE: A prospective investigation of the postoperative intraocular pressure level, visual function changes, and postoperative complications of trabeculectomy with adjunctive mitomycin C in normal-tension glaucoma. PATIENTS AND METHODS: Thirty-one patients (31 eyes) who underwent trabeculectomy with 0.4 mg/ml mitomycin C were studied. The preoperative intraocular pressure of the eyes ranged from 11 to 18 mm Hg with a mean (+/- SD) of 14.1 +/- 1.9 mm Hg. Follow-up examinations were performed at least once a month for 14-35 months (mean: 24.4 months). RESULTS: Intraocular pressures decreased to between 4 and 14 mm Hg with a mean (+/- SD) of 8.4 +/- 2.4 mm Hg at the last three follow-up visits. The magnitude of the intraocular pressure reduction ranged from 1 to 10 mm Hg, with a mean of 5.8 mm Hg. Final measurements of postoperative intraocular pressure ranged between 5 and 12 mm Hg in 27 eyes (87%). Visual acuity deteriorated two lines or more in seven eyes, which was caused by cataract. The visual field deteriorated postoperatively in two eyes (6%): their respective postoperative intraocular pressures averaged 9.4 and 7.5 mm Hg. Hypotonous maculopathy developed in three eyes (10%). CONCLUSIONS: Trabeculectomy with mitomycin C appears to have potential as a treatment modality of choice for normal-tension glaucoma, because a large majority of the patients in the present study achieved an intraocular pressure allegedly beneficial in managing normal-tension glaucoma.

We examined 101 patients (171 eyes) with primary angle-closure glaucoma by pressure gonioscopy to study the distribution and the morphology of peripheral anterior synechiae. The incidence of peripheral anterior synechiae was 86.8% in the eyes with acute primary angle-closure glaucoma, 51.4% in their fellow eyes, and 83.3% in the eyes with chronic primary angle-closure glaucoma. The peripheral anterior synechiae in the eyes with acute primary angle-closure glaucoma were wider and taller than those found in the eyes with chronic primary angle-closure glaucoma. Wide peripheral anterior synechiae were most prevalent in the superior sector and were, overall, more prevalent than the narrow type. Peripheral anterior synechiae, as a whole, were most frequently seen in the superior sector. The wide peripheral anterior synechiae were taller than the narrow type. The age of the patient and the extent of the peripheral anterior synechiae (peripheral anterior synechiae index) showed no significant correlation. A significant correlation (r = 0.68, p < 0.01) was found between the duration of acute attacks and the peripheral anterior synechiae index.