Biomedical subjects

H Ohtake

Publications and source records attributed to H Ohtake.

At least 55 records · Page 3Linked to original sources

PubMed1999

Cloning and characterization of Pseudomonas putida genes encoding the phosphate-specific transport system.

The pstSCAB genes of Pseudomonas putida PRS2000, encoding the phosphate (Pi)-specific transport (Pst) system, were cloned. The pstS gene of Pseudomonas aeruginosa PAO1, of which the pstCAB genes had been cloned previously, was also cloned (Nikata, T. et al., Mol. Gen. Genet., 250, 692-698, 1996). The predicted translation products of the P. putida pstSCAB genes showed 83, 75, 78 and 88% amino acid identity with their P. aeruginosa counterparts. Two well-conserved Pho box sequences were found in the region upstream of the pstS gene (15/18 base identity with the consensus Pho box sequence) and in the intercistronic region between the pstS and pstC genes (11/18 base identity) of P. putida PRS2000. To investigate the functions of PstSCAB, the pstSC genes were inactivated by inserting a kanamycin resistance gene cassette into the chromosome of P. putida PRS2000. The resultant mutant, designated PNT1, failed to take up 32Pi even under conditions of Pi limitation. Strain PNT1 was also constitutive for alkaline phosphatase synthesis, as well as chemotaxis toward Pi, indicating that the Pst system is involved in the negative regulation of the pho regulon in P. putida. Although overexpression of the pstSCAB genes in P. putida PRS2000 resulted in decreased cell growth, this recombinant strain could remove Pi at a rate similar to that seen with the control strain.

Journal Article↗

PubMed1999

In vitro and in vivo evaluation of the enhancing activity of glycyrrhizin on the intestinal absorption of drugs.

PURPOSE: The enhancing activity of dipotassium glycyrrhizinate (Grz) on the intestinal absorption of drugs has been demonstrated in an in vitro study using Caco-2 cell monolayers and in an in vivo absorption study in rats. METHODS: The hydrolysis of Grz by luminal content and mucosa of the rat colon was investigated. The absorption-enhancing activity of Grz and its hydrolysates was estimated by changes in transepithelial electrical resistance (TEER) and the permeation of sodium fluorescein (Flu-Na) in Caco-2 cell monolayers. It was further evaluated through the absorption of salmon calcitonin (sCT) in the rat colon. RESULTS: Grz was not hydrolyzed to glycyrrhetinylmonoglucuronide (GrMG) and glycyrrhetinic acid (GA) by colonic mucosa, but, rather by the beta-glucuronidase in colonic flora. The hydrolysis of Grz to GrMG was extremely slow and the GrMG produced was rapidly regenerated to GA. Grz and GrMG had no effect on TEER nor on the permeability of Flu-Na across Caco-2 cell monolayers. On the other hand, GA decreased TEER and increased the permeability of Flu-Na in a dose-dependent manner. However, Grz and GrMG enhanced the plasma calcium-lowering effect of sCT after administration in the rat colon. The coadministration of sCT and GA in the rat colon induced the strongest plasma calcium-lowering effect and the highest plasma concentration of sCT. CONCLUSIONS: The in vivo enhancing-activity of Grz in the absorption of drugs is dependent on GA, a hydrolysis product of Grz resulting from the action of beta-glucuronidase in intestinal flora.

Animals↗

PubMed1999

Purification and characterization of prolyl endopeptidase from the Pacific herring, Clupea pallasi, and its role in the activation of sperm motility.

Protease activities with specificity toward synthetic substrates, Suc-Gly-Pro-Leu-Gly-Pro-MCA for prolyl endopeptidase or collagenase-like peptidase, and Suc-Ala-Ala-Pro-Phe-MCA for chymotrypsin were identified in the detergent-soluble fraction of herring spermatozoa. The enzyme activities increased in the presence of herring sperm-activating protein (HSAP). Among them a prolyl endopeptidase [EC. 3. 4. 21. 26] was purified to near homogeneity from herring testis. The molecular mass of the enzyme was 79 kDa and the properties of the enzyme were quite similar to prolyl endopeptidase from other tissues or cells. Both the enzyme activation and the sperm motility activation by HSAP were inhibited by benzyloxycarbonyl-L-thioproline-thioprolinal, a specific inhibitor for prolyl endopeptidase. Furthermore, the motility activation by HSAP was inhibited by substrates of the prolyl endopeptidase. Western blotting with mouse anti-prolyl endopeptidase serum revealed the presence of 79 kDa prolyl endopeptidase in the tail fraction of herring sperm. These results suggest that prolyl endopeptidase exists on the surface of the sperm tail and interacts with the HSAP.

Amino Acid Sequence↗

PubMed1999

Arterial anastomosis using the telescoping method and an elastomeric adhesive.

We experimentally studied a new technique for anastomosis of small arteries which involves the telescoping method and a surgical adhesive with the objective of examining its clinical potential. This technique was applied to the unilateral femoral arteries of 27 mongrel dogs. After division of the artery, the distal artery was incised longitudinally, and the proximal end was invaginated into the opened artery. One stay suture and the elastomeric adhesive PUP201 were placed in the anastomotic site. The bursting and tensile strength and the patency were examined, and a histopathological study was performed at various intervals up to 1 year after the operation. The bursting strength exceeded 500 mm Hg. The patency rate of the anastomosed arteries was 100%. The mean percentage of stenosis to diameter of the proximal normal lumen was 20% one year after operation. The luminal surface of the anastomosed line was healed smoothly, and thrombi between the telescoped arteries were replaced by elastofibrotic union. In conclusion, small arterial anastomosis using the telescoping method and an elastomeric adhesive is easy and safe and provides good patency.

Anastomosis, Surgical↗

PubMed1999

Isolation and characterization of two cryptic plasmids in the ammonia-oxidizing bacterium Nitrosomonas sp. strain ENI-11.

Two plasmids were discovered in the ammonia-oxidizing bacterium Nitrosomonas sp. strain ENI-11, which was isolated from activated sludge. The plasmids, designated pAYS and pAYL, were relatively small, being approximately 1.9 kb long. They were cryptic plasmids, having no detectable plasmid-linked antibiotic resistance or heavy metal resistance markers. The complete nucleotide sequences of pAYS and pAYL were determined, and their physical maps were constructed. There existed two major open reading frames, ORF1 in pAYS and ORF2 in pAYL, each of which was more than 500 bp long. The predicted product of ORF2 was 28% identical to part of the replication protein of a Bacillus plasmid, pBAA1. However, no significant similarity to any known protein sequences was detected with the predicted product of ORF1. pAYS and pAYL had a highly homologous region, designated HHR, of 262 bp. The overall identity was 98% between the two nucleotide sequences. Interestingly, HHR-homologous sequences were also detected in the genomes of ENI-11 and the plasmidless strain Nitrosomonas europaea IFO14298. Deletion analysis of pAYS and pAYL indicated that HHR, together with either ORF1 or ORF2, was essential for plasmid maintenance in ENI-11. To our knowledge, pAYS and pAYL are the first plasmids found in the ammonia-oxidizing autotrophic bacteria.

Amino Acid Sequence↗

PubMed1999

Coelomic metaplasia theory of endometriosis: evidence from in vivo studies and an in vitro experimental model.

Ultrastructure studies of pelvic peritoneal tissue from women undergoing laparotomy suggest that before endometriosis has become established in the peritoneum, there might be a metaplastic change by peritoneal mesothelial cells into endometrial glandular cells. A new in vitro experimental model of endometriosis using human ovarian surface epithelium cells has shown evidence that endometriotic lesions can arise by a process of metaplasia from the ovarian surface epithelium. In this model, when both ovarian surface epithelium and ovarian stromal cells were cocultured with 17beta estradiol in a three-dimensional collagen gel lattice, the ovarian surface epithelium cells formed a lumen structure, surrounded by endometrial stromal cells with an epithelial mesenchymal structure. Immunoreactivity for epithelial membrane antigen and cytokeratin was shown in the glandular cells and cilia, as well as in the microvilli. Electron microscopy showed evidence of tight junctions on cell surfaces. These findings suggest that endometriosis may manifest as a serial change from the adjacent mesothelial cells.

Cells, Cultured↗

PubMed1999

Simultaneous use of sodium deoxycholate and dipotassium glycyrrhizinate enhances the cellular transport of poorly absorbed compounds across Caco-2 cell monolayers.

The absorption-enhancing effect of a combination of sodium deoxycholate and dipotassium glycyrrhizinate in Caco-2 cell monolayers has been compared with that of the enhancers when used alone, and the mechanism of the enhancement was partially elucidated. The effect of the combined compounds was evaluated by measurement of transepithelial electrical resistance (TEER) and the cellular permeability of the water-soluble model compounds sodium fluorescein (MW 376.3) and fluorescein isothiocyanate dextran (MW 4400). The TEER of the monolayers decreased with increasing concentrations of dipotassium glycyrrhizinate in combination with 0.02% (w/v) sodium deoxycholate for 20 min, and reached a minimum at 1% (w/v) dipotassium glycyrrhizinate. Although a combination of 0.02% (w/v) sodium deoxycholate and 1% (w/v) dipotassium glycyrrhizinate enhanced the cellular permeability of sodium fluorescein and fluorescein isothiocyanate dextran, 0.02% (w/v) sodium deoxycholate and 1% (w/v) dipotassium glycyrrhizinate alone had no effect on either the TEER of the monolayers or the cellular transport of the water-soluble compounds. Sequential and separate exposure of the monolayers to each enhancer for 10 min had no effect on the TEER, but a marked decrease in TEER was observed when both compounds were used in combination. The enhancing effect of the combination of sodium deoxycholate and dipotassium glycyrrhizinate was inhibited by H7, a protein kinase C (PKC) inhibitor, suggesting that dipotassium glycyrrhizinate might enhance the activation of PKC via sodium deoxycholate. The combined use of these two enhancers had no toxic effects. These results provide useful, basic information on the action of these absorption enhancers on drugs for which absorption is limited owing to polarity or molecular size, or both.

Absorption↗

PubMed1999

Cloning and characterization of chemotaxis genes in Pseudomonas aeruginosa.

Two chemotaxis-defective mutants of Pseudomonas aeruginosa, designated PC3 and PC4, were selected by the swarm plate method after N-methyl-N'-nitro-N-nitrosoguanidine mutagenesis. These mutants were not complemented by the P. aeruginosa cheY and cheZ genes, which had been previously cloned (Masduki et al., J. Bacteriol., 177, 948-952, 1995). DNA sequences downstream of the cheY and cheZ genes were able to complement PC3 but not PC4. Sequence analysis of a 9.7-kb region directly downstream of the cheZ gene found three chemotaxis genes, cheA, cheB, and cheW, and seven unknown open reading frames (ORFs). The predicted translation products of the cheA, cheB, and cheW genes showed 33, 36, and 31% amino acid identity with Escherichia coli CheA, CheB, and CheW, respectively. Two of the unknown ORFs, ORF1 and ORF2, encoded putative polypeptides that resembled Bacillus subtilis MotA (40% amino acid identity) and MotB (34% amino acid identity) proteins, respectively. Although P. aeruginosa was found to have proteins similar to the enteric chemotaxis proteins CheA, CheB, CheW, CheY, and CheZ, the gene encoding a CheR homologue did not reside in the chemotaxis gene cluster. The P. aeruginosa cheR gene could be cloned by phenotypic complementation of the PC4 mutant. This gene was located at least 1,800 kb away from the chemotaxis gene cluster and encoded a putative polypeptide that had 32% amino acid identity with E. coli CheR.

Amino Acid Sequence↗

PubMed1999

Exclusion of internal iliac arterial aneurysm concomitant with abdominal aortic aneurysm repair.

BACKGROUND: To study the long-term outcomes after exclusion of internal iliac arterial aneurysm performed concomitantly with abdominal aortic aneurysm repair in patients with ruptured aortic aneurysm or other high-risk conditions. METHODS: The 31 patients who participated in this study underwent emergency (N = 9) or elective surgery (N = 22). The abdominal aortic aneurysm and the common iliac artery were excluded together with the internal iliac aneurysm in 7 patients. Forty-three (12 bilateral and 19 unilateral) internal iliac aneurysms were excluded: 35 by proximal ligation only, 5 by proximal and distal ligation, and 3 by partial resection of the proximal part of the aneurysm. The platelet count and fibrinogen level were evaluated pre- and postoperatively. Pelvic organ ischemia, classed as ischemic colitis, buttock claudication and sexual dysfunction, was examined. RESULTS: The inferior mesenteric artery was reimplanted in 21 patients. The platelet count dropped significantly postoperatively, but the fibrinogen level increased and no bleeding tendency was noted. Ischemic colitis occurred in 7 patients, resulting in colonic infarction in 2 patients. The operative mortality was 16%, and the postoperative observation periods ranged from 4 days to 217 months (mean, 60 months). The incidence of buttock claudication and sexual dysfunction was 12% and 39%, respectively. The excluded aneurysms were all thrombosed at discharge, and no late rupture was noted. The 5- and 10-year survival rate after surgery was 56% and 51%, respectively. CONCLUSIONS: Exclusion of the internal iliac aneurysm concomitant with abdominal aortic aneurysm repair shows acceptable outcome when performed in patients with high-risk conditions.

Aged↗

PubMed1999

[Two cases of fluent aphasia with selective difficulty of syllable identification].

We report two aphasic patients who could discriminate Japanese syllables but could not identify them. Case 1 was a 51-year-old right handed woman with 12-year education. Case 2 was a 50-year-old right handed man with 9-year education. They developed fluent aphasia after a cerebral infarction. Brain MRI of case 1 revealed widely distributed lesions including inferior frontal, superior temporal, angular and supramarginal gyri. Lesions revealed by Brain CT in case 2 included the left superior and middle temporal, angular and supramarginal gyri. Both showed severe impairment of repetition and confrontation naming. No difference of performance was present between repetition of single syllables and polysyllabic words. On the contrary, oral reading of Kana characters were preserved. We examined their ability to perceive syllables in detail. In the discrimination task, they judged whether a pair of heard syllables was same or different. Case 1 was correct in 85% of the tasks and case 2 in 98%. In an identification task, they heard a syllable and chose a corresponding Kana, Kanji, or picture out of 10 respective candidates. Case 1 was correct only in 30% and case 2 in 50% of these tasks. On the other hand, selection of a correct target in response to a polysyllabic word was much better, i.e. 70% in case 1 and 90% in case 2. Based on these data we concluded that (1) syllabic identification is a different process from syllabic discrimination, and (2) comprehension of a polysyllabic word can be achieved even when the precise phonological analysis of continuent syllables are impaired.

Aphasia, Wernicke↗

PubMed1998

Sperm-activating proteins obtained from the herring eggs are homologous to trypsin inhibitors and synthesized in follicle cells.

The activation of sperm motility by the egg is an ubiquitous phenomenon in the animal kingdom, but the molecules by which the egg activates sperm motility have been clarified in only a few invertebrate species. In the Pacific herring, Clupea pallasii, mature unfertilized eggs release the sperm-activating proteins which are prerequisite to successful fertilization. Complementary DNA clones encoding herring sperm-activating proteins were isolated from a herring ovarian complementary DNA library and amino acid sequences were deduced. The herring sperm-activating protein(s) is a secretory product(s) with a strong homology to Kazal-type trypsin inhibitors, such as mammalian acrosin inhibitors. The sperm-activating proteins were globally distributed in the outermost layer of the egg chorion and its gene was found to be expressed in the follicle cells which surround developing oocytes. These results suggest that in the Pacific herring, trypsin inhibitor-like proteins are synthesized in the follicle cells, secreted, accumulated in the egg chorion during oocyte development, and released into the milieu at spawning to activate the motility of spermatozoa at the time of gamete interaction.

Amino Acid Sequence↗

PubMed1998

Larynx-preserving resection of the cervical esophagus for cervical esophageal carcinoma limited to the submucosal layer.

This report describes the surgical procedure consisting of larynx-preserving resection of the cervical esophagus and satisfactory lymphadenectomy. The sternum was split at the level of the 3rd intercostal space, which allowed an upper-mediastinal lymphadenectomy to be performed easily. The cervical esophagus was reconstructed using a free jejunal autograft. The stump of the thoracic esophagus and the caudad stump of the jejunal graft were anastomosed using a circular stapling instrument. The posterior part of the cephalad esophagojejunostomy was completed in two layers using the Lembert stitch. The wall of the cervical esophagus was opened to determine the oral cut line considering the safety margin from the carcinoma. After cervical esophagectomy was completed, suturing of the anterior wall was performed in one layer. The left cervical transverse artery and the internal jugular vein were employed for recipient vessels. This procedure is acceptable for high cervical esophageal carcinoma limited to the submucosal layer.

Carcinoma, Squamous Cell↗

PubMed1998

Establishment of a new cell line, OKT1, from small cell carcinoma secreting ectopic ACTH of the uterine cervix.

OBJECTIVE: Small cell carcinoma of the uterine cervix is rare and represents a unique entity among gynecological tumors. It sometimes demonstrates neuroendocrine differentiation, including adrenocorticotropin (ACTH) secretion. In this study, we established a new cell line, OKT1, from a case of carcinoma secreting ectopic ACTH without Cushing's syndrome and determined the character of the cell line. METHODS: OKT1 was established from OKT tumor cells, derived from a biopsy specimen of small cell cervical carcinoma, and serially heterotransplanted into nude mice. To characterize OKT1, the cell morphology, growth properties, immunohistochemical properties, hormone- and tumor-associated antigen secretion, tumorigenic potential, DNA profile, and chromosomal alteration were studied. RESULTS: The population doubling time of OKT1 was approximately 27 h. The cytological properties of OKT1, including DNA ploidy pattern, were similar to those of the primary tumor. Neuroendocrine differentiation was shown in the OKT1 cells by the positive immunocytochemical staining of neuron-specific enolase (NSE) and the presence of NSE and ACTH in the culture media. The xenograft of 1 x 10(8) OKT1 cells into nude mice yielded tumor mass. Furthermore, OKT1 demonstrated HPV type 18 and absence of a p53 gene mutation from exons 5 through 8. CONCLUSION: To our knowledge, OKT1 is the first cell line established from small cell cervical carcinoma with ACTH secretion.

Adrenocorticotropic Hormone↗

PubMed1998

Surgical Procedure for Right Anteroseptal Accessory Conduction Pathways in Wolff-Parkinson-White Syndrome.

In patients with Wolff-Parkinson-White syndrome, the right anteroseptal accessory conduction pathway is rare, and exists from the atrium to the ventricle in close anatomic proximity to the normal atrioventricular conduction system. Catheter ablation of this lesion is reported to interrupt atrioventricular node-His bundle conduction more easily than that of other lesions. At our institute, there were 10 patients with right anteroseptal accessory conduction pathway among 454 patients (2.2%) who underwent the surgical division of the accessory pathway. Our procedure involved the the endocardial approach with knife dissection and cryocoagulation. With the heart beating under normothermal cardiopulmonary bypass, delta wave disappearance was easily noted. Neither complete atrioventricular block nor recurrent conduction occurred. Cryoablation used by our endocardial surgical division, is a safe and accurate procedure.

Journal Article↗

PubMed1998

Long-term results of endarterectomy, anatomic bypass and extraanatomic bypass for aortoiliac occlusive disease.

Over the past 20 years, 214 patients in our hospital were operated on for aortoiliac occlusion, including an endarterectomy for 88 legs in 64 patients, an anatomic bypass for 178 legs in 105 patients, and an extraanatomic bypass for 51 legs in 45 patients. The extraanatomic group was older than the other two groups (70 years versus 64 years), and also showed a higher instance of critical ischemic symptoms than the other groups (49% versus 16%). The 10-year primary patency rate was 88% for an endarterectomy, 91% for an anatomic bypass, and 73% for an extraanatomic bypass. The 10-year survival rate was 71% for an endarterectomy, 49% for an anatomic bypass, and 24% for an extraanatomic bypass. The possible complications related to each surgical method included sexual dysfunction resulting from endarterectomy, and graft infection and anastomotic aneurysm in the prosthetic bypass. No ischemic symptoms in the donor limb were noted following an extraanatomic bypass. The selection of the appropriate surgical method for aortoiliac occlusion should thus be made after a careful review of the patient's general condition, the morphology of arterial occlusion, and the risk of possible complications for each type of surgery.

Adult↗

PubMed1998

Surgical drainage for idiopathic suppurative pylephlebitis.

Idiopathic suppurative pylephlebitis is quite rare and only a few cases have been reported. Conservative systemic administration of antibiotics and urokinase is reported to be effective. In this report, surgical drainage was performed on an 18-year-old man who complained of fever and abdominal pain. He had no past history of abdominal inflammatory disease or abdominal surgery. Ultrasonography and computed tomography showed wide spread thrombosis of the portal vein. Laparotomy was performed and the occluded superior mesenteric vein was incised. Massive pus was removed. Thereafter, a drain was placed at the opened mesenteric vein. Drainage resulted in a dramatic decrease in fever. Postoperative radiographic studies of the colon, the small intestine, and other organs did not show any abnormalities. Emergency surgical drainage was performed successfully, instead of systemic administration of antibiotics and urokinase. Surgical drainage may be useful for wide spread pylephlebitis and pylethrombosis.

Abdomen, Acute↗

PubMed1998

Effects of absorption enhancers on cytoskeletal actin filaments in Caco-2 cell monolayers.

Histomorphological changes of actin filaments, intracellular levels of calcium ion, and amount of released lactate dehydrogenase (LDH) were examined, in order to elucidate the mechanism of action of three absorption enhancers, i.e., sodium caprate (Cap-Na), sodium deoxycholate (Deo-Na), and dipotassium glycyrrhizinate (Grz-K), using Caco-2 cell monolayers. The structure of actin filaments in microvilli was slightly modified by 0.5 %(w/v) Grz-K and was significantly changed by 0.2 %(w/v) Cap-Na, 0.05 %(w/v) Deo-Na, and 0.0015 %(w/v) ionomycin. All of the enhancers, except Grz-K, induced significantly histomorphological changes in the actin filaments on the middle depth and basal side of the cells. Furthermore, the altered structure of the actin filaments in the monolayers was restored after removal of the Cap-Na, Grz-K and ionomycin, but not Deo-Na. Intracellular levels of calcium ion increased in the following order: ionomycin = Cap-Na > Deo-Na. However, the intracellular calcium ion levels decreased by treatment with Grz-K. The changes in transepithelial electric resistance (TEER) at the initial stage of treatment with all enhancers correlated with intracellular calcium ion levels. These results suggest that one of the mechanisms by which these agents exert absorption-enhancing activity involves structural alterations in the cytoskeletal actin filaments which are provoked by changes in intracellular calcium ion levels. Only the monolayers which were treated with 0.05 %(w/v) Deo-Na released a significant amount of LDH and irreversibly altered the structure of actin filaments, thus indicating that Deo-Na might affect the actin filaments not only by increasing intracellular calcium ion level but also by other, presently unknown factors.

Actins↗

PubMed1998

Prevention of spinal cord ischemia by selective intercostal arterial infusion of prostaglandin E1.

PURPOSE: A new protective method against the spinal cord ischemia that occurs during aortic clamping was investigated in dogs. Oxygenated blood containing prostaglandin E1 (PGE1) was administered at the clamped aortic segment, and the effect was evaluated by measurement of the sensory evoked spinal potential (SESP). METHODS: In 30 dogs, a thoracotomy was made with dissection of the thoracic aorta. After intravenous heparin (100 units/kg) was administered, the proximal and distal descending thoracic aortas were cross-clamped for 60 minutes. Group A (n=10) received oxygenated blood at the rate of 1.0 ml/kg/min. Groups B (n=10) and C (n=10) received oxygenated blood at the same rate, with PGE1 at the dosage of 25 and 50 ng/kg/min, respectively. The infusion was continuously administered throughout the entire period of ischemia. SESP was measured with epidural electrodes before clamping, 10 and 60 minutes after clamping, and 10 and 60 minutes after declamping. Neurologic outcome was assessed at 24 hours after the operation and graded according to the method of Tarlov. RESULTS: There was no significant hemodynamic change in any group. At 60 minutes after damping and at 10 and 60 minutes after declamping, the amplitude of SESP was lower than that at preclamping in groups A and B (p < 0.05). At 60 minutes after damping and at 10 and 60 minutes after declamping, the SESP was more markedly decreased in group A compared with groups B and C. Regarding postoperative neurologic outcome, the dogs with SESP amplitude of more than 50% of the preclamping control value at 60 minutes after clamping showed neither paralysis nor paraplegia. Seven of nine dogs with less than 50% SESP amplitude showed neurogenic deficit. In a comparison of groups A, B, and C, the Tarlov score for group A dogs was significantly lower than that for group C dogs (p < 0.05). CONCLUSION: In this model, PGE1 administration at the rate of 50 ng/kg/min showed sufficient spinal cord protection against ischemia without a decrease in the blood pressure. Further studies are needed to determine the dose that will provide the maximal protective effect and to determine the maximum duration of ischemia against which PGE1 shows protective effects.

Alprostadil↗