Bradykinin receptor antagonists attenuate neointimal proliferation postangioplasty.

Bradykinin has been linked to the development of restenosis in response to vascular injury. We therefore examined the effect of bradykinin on vascular smooth muscle cell growth and neointimal formation in organ culture. Bradykinin stimulated both RNA and DNA synthesis (by 175%) in smooth muscle cells from either porcine or human coronary arteries and increased cell number in a concentration-dependent manner. Both p42/44 mitogen-activated protein kinase (MAPK) and p38 kinase were also activated. Treatment with [Hyp(3),Tyr(Me)(8)]bradykinin, a B(2) receptor agonist, stimulated thymidine incorporation by 146%, whereas B(1)-selective Lys-des-Arg(9)-bradykinin had no effect. Addition of the B(2) antagonist HOE-140 reduced the stimulation by 56%, whereas B(1)-selective des-Arg-HOE-140 had no significant effect. Similarly, HOE-140 attenuated angioplasty-induced neointimal formation in organ culture with an efficacy approaching 100% inhibition. These experiments suggest that bradykinin promotes smooth muscle proliferation after vascular injury, presumably via B(2) receptor-dependent activation of MAPK family pathways, and may explain the negative outcome of angiotensin converting enzyme inhibitor therapy on restenosis in nonrodent models.

Sarpogrelate inhibits serotonin-induced proliferation of porcine coronary artery smooth muscle cells: implications for long-term graft patency.

BACKGROUND: Serotonin can induce proliferation of vascular smooth muscle cells. We assessed the ability of a specific serotonin receptor antagonist, sarpogrelate, to inhibit proliferation of cultured porcine coronary artery smooth muscle cells. METHODS: Cell proliferation and mitotic activity were measured using 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyltetrazolium bromide. To determine the effect of sarpogrelate on DNA (deoxyribonucleic acid), RNA (ribonucleic acid), and protein synthesis, radioactive incorporation of 3H-thymidine, 3H-uridine, and 3H-phenylalanine, respectively, was used. Synthesis of DNA was also assessed by flow cytometry with propidium iodide as a fluorochrome. RESULTS: Serotonin, platelet-derived growth factor, endothelin, and angiotensin II all induced proliferation of porcine coronary artery smooth muscle cells. Sarpogrelate specifically inhibited the serotonin-induced cytokine trigger but did not influence platelet-derived growth factor-, endothelin-, or angiotensin II-induced cell proliferation. Sarpogrelate inhibited the serotonin-induced increase in intracellular free ionized calcium concentration, prevented mitogen-activated protein kinase activation, and down-regulated expression of the protooncogenes c-fos and c-jun. Sarpogrelate acted at the G1 phase of the cell cycle. CONCLUSIONS: These data suggest that sarpogrelate could be used as a therapeutic agent to inhibit serotonin-induced neointimal hyperplasia and improve patency of coronary artery bypass grafts.

A novel organ culture method to study intimal hyperplasia at the site of a coronary artery bypass anastomosis.

BACKGROUND: Intimal hyperplasia or restenosis at the site of a coronary artery bypass anastomosis contributes to early graft failure, and growth factor release in response to construction of the anastomotic site strongly influences this process. Due to the difficulties in studying restenosis after coronary artery bypass graft surgery, we have tested whether an organ culture model we have developed can simulate the early events associated with intimal hyperplasia. METHODS: End-to-side anastomosis of porcine radial artery to porcine coronary artery were constructed. The vessels were trimmed and incubated under standard tissue culture conditions for 14 days. Appropriate controls were treated similarly. The vessels were frozen, cryosectioned, and immunostained for the expression of the proliferation marker proliferating cell nuclear antigen (PCNA). A proliferative index (PCNA positive nuclei/total nuclei) was calculated for comparative purposes. RESULTS: Limited PCNA staining was observed in noncultured vessel segments (0.046+/-0.045). A slight increase in this index was observed in vessels that had been placed into culture without manipulation (0.230+/-0.141) and in vessels subjected to an arteriotomy (0.462+/-0.249). However, the most significant increase was obtained after construction of an anastomosis (4.98+/-6.66). No change in total cell number was evident over the course of the experiment or in relation to the treatment. CONCLUSIONS: Culture conditions and incision slightly stimulate cell proliferation in porcine coronary artery segments when compared with basal conditions of a native artery. In contrast, construction of an anastomosis increases proliferation 108-fold. Therefore, surgical manipulation of arterial conduits during construction of an anastomotic site is the primary trigger for intimal hyperplasia, independent of dissection and incision of the vessel. Furthermore, these data indicate the organ culture model we have developed will be useful for examining the cellular and molecular mechanisms that mediate intimal hyperplasia at the site of a coronary artery bypass graft anastomosis.

Cardiac collagen remodeling in the cardiomyopathic Syrian hamster and the effect of losartan.

Although increased deposition of collagen proteins has been described in cardiomyopathy, little is known of the temporal relationship between events in collagen gene transcription and the occurrence of cardiac fibrosis, the removal of collagen by matrix metalloproteinases (MMPs), or of the regulation of these events by angiotensin AT1 receptors in this disease. We sought to study steady-state collagen mRNA abundance and the deposition of specific collagen subtypes in right and left ventricular muscle of Syrian cardiomyopathic (CMP) hamsters at different stages of cardiomyopathy. Using zymography, we also investigated the gelatinolytic activities of different MMPs to gain some information about collagen removal in experimental hearts. Finally, we investigated the effect of AT1 receptor blockade (losartan) on collagen remodeling. We observed that the mRNA levels of types I and III collagens were significantly increased in all four experimental groups (35, 65, 120, and 200 day) in left ventricular tissue when compared to control (F1-beta strain) values. The mRNA levels of these collagen species in experimental right ventricular tissue samples were only elevated significantly in the 35 and 200 day experimental groups when compared to controls. Fibrillar collagen deposition was elevated in left and right ventricular CMP samples after a lag period from the occurrence of corresponding increases in mRNA abundance. Although 2-week losartan treatment of 65, 120 and 200 day experimental groups had no significant effect on left ventricular fibrillar collagen concentration or collagen mRNA abundance when compared to vehicle-infused CMP hamsters, AT1 receptor blockade was associated with complete regression of cardiac hypertrophy. Both MMP-1 (54 kDa band) and MMP-2 (58 and 62 kDa bands) activities were increased in left ventricular CMP tissues at 65, 120 and 200 days when compared to F1-beta controls. Losartan treatment was associated with significant attenuation of MMP activities in cardiomyopathic samples at 65 and 120 days. Thus, elevation of mRNA abundance of fibrillar collagen genes occurs at very early stages in this model of cardiomyopathy, and corresponding collagen proteins were subsequently deposited in the cardiac interstitium at later stages. As collagen concentration was significantly increased in later stages of cardiomyopathy studied herein (120 and 200 day groups), our data support the hypothesis that collagen synthesis exceeds the capacity of collagen removal during the progression of cardiomyopathy. Nevertheless, cardiac collagen remodeling may be facilitated by elevated MMP activity in cardiomyopathic stages in this experimental model, and we suggested that attenuation of MMP activity in the presence of losartan may be a cardioprotective mechanism of this agent.

Soliciting and evaluating quality-based bids.

This example clearly indicates that the further application of this methodology within the hospital industry is both useful and beneficial: useful as an aid in selecting the best product, and beneficial to the hospital and consumers because, in many cases, it may serve to lower the costs associated with the procurement of these products. Although the methodology itself is quite simple to use, the impact of this methodology can be far reaching. This experience also further supports the need for the adaptation of techniques within the hospital industry from other fields and from other industries.

[Percutaneous transluminal revascularization of lower-leg arteries]. / Percutane transluminale Revascularisation von Unterschenkelarterien.

In 19 of more than 130 patients treated with percutaneous transluminal angioplasty (PTA), obstructions of the distal popliteal artery and the lower leg arteries encouraged us to extend the treatment to this vascular region. Two weeks after PTA the arteries were patent in 16 of the 19 cases (84%). These results are comparable with those of vascular surgery, but are gained with less injury to the patient and a shorter hospitalization time.

[Morbus Wilson--pathogenesis, diagnosis, therapy, and course (author's transl)]. / Morbus Wilson--Pathogenese, Diagnose, Therapie und Verlauf.

During childhood, Wilson's disease becomes manifest mostly in the hepatic form. In children every case of cirrhosis of the liver, hemolysis with high levels of conjugated bilirubin in the serum, and otherwise in explicable tremor make it imperative to exclude or confirm the existence of Wilson's disease. A false diagnosis often delays the start of therapy with d-penicillamine and low-copper diet. The prognosis, which was still fatal a few years ago, has improved considerably thanks to new therapeutic possibilities.

[Morbus wilson--progressive fatal course of the hepatic form with apoplexy and abscess of the brain (author's transl)]. / Morbus Wilson--rasch progredienter Verlauf bei einer hepatischen Form mit Apoplexie und Hirnabszess.

The onset with a hemolytic crisis, as often described in the literature, led to the diagnosis of Wilson's disease. Treatment was immediately started in the usual way with diet and d-penicillamin. But the disease had already caused serious liver damage and renal lesions, and the patient died fourteen weeks after diagnosis. Examination of further family members disclosed a cousine with impaired liver and kidney function, also suffering from the hepatic form of homozygous Wilson's disease.