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Proteolytic enzymes have shown efficacy in cancer therapy. We present a combination of the two pro-enzymes Trypsinogen and Chymotrypsinogen A with potent in vitro and in vivo anti-tumour efficacy. A synergetic anti-tumour effect for Trypsinogen and Chymotrypsinogen A was determined at a ratio 1:6 (named PRP) using 24 human cancer cell lines. The antiangiogenic effect of PRP was analysed by matrigel-based tube formation and by fibrous capsule formation assays. Furthermore, cell invasion and wound healing assays together with qRT-PCR determination of epithelial-to-mesenchymal transition (EMT) markers were performed on human cancer cells treated with PRP. Additionally, in vivo pharmacokinetic studies were implemented and the PRP's anti-tumour efficacy was explored against orthotopic pancreatic and ovarian cancer tumours. PRP formulation was proven to inhibit in vitro angiogenesis, tumour growth, cancer cell migration and invasiveness; and to be an effective and well tolerated in vivo anti-tumour treatment. Finally, the clinical efficacy of a suppository formulation containing both pancreatic pro-enzymes in the context of a UK Pharmaceuticals Special Scheme was evaluated in advanced cancer patients. Consequently, PRP could have relevant oncological clinical applications for the treatment of advanced or metastatic pancreatic adenocarcinoma and advanced epithelial ovarian cancer.

It is imperative to maintain normal blood flow to provide adequate oxygen supply to specific organs and cells, as well as for the removal of metabolic byproducts. Therefore, any situation that results in blood clotting can injure or kill living tissues. In this paper, we describe a case where a protein C deficient subject who would, by all medical indicators, be at 100 % risk of experiencing thrombophlebitis, deep vein thrombosis, and or lung emboli, is able to escape all pathologies by using perioperative zymogen protein C (ZPC). This protein C deficient patient has a long history of blood clotting, particularly from surgical procedures. The patient is 81 years old and first experienced clotting due to hernia surgery in 1964, when he was hospitalized for 16 days post-surgery with life threatening complications. It was later determined in 1980, after many episodes, that the patient had hereditary protein C deficiency at the 38 % level. In his hernia surgery, perioperative ZPC was used along with accepted anticoagulation procedures with no blood clots or other related side effects occurring. This procedure can greatly benefit protein C deficient patients, and could potentially find use for non-PC deficient patients in surgeries and a variety of other medical treatments. This particular case helps to validate the importance of ZPC in effecting safer surgery in high-risk patients. It also supports the mechanism of ZPC acting as an anticoagulant without causing bleeding. Most importantly, each clinical case study represents a unique combination of surgeon, hematologist, medical staff, and patient functioning as a coordinated team. In this case, smaller amounts of very expensive ZPC achieved safe and efficacious results, which is hugely important for future clinical applications when considering the production cost of ZPC. More studies must be done to establish minimum dosing while achieving safe and efficacious outcomes.

The blood clotting mechanism is a very important and complex physiologic process. Blood flow must be continuous through the blood vessels to provide essential oxygen and nutrients to the cells of the body. Dr. Melvin H. Knisely (Honorary First President of ISOTT, 1973) named and pioneered research in blood sludging and clotting which led to his nomination for the Nobel Prize by Dr. August Krogh in 1948. Abnormal clotting is a pathological state that can inhibit and prevent normal blood flow, leading to reduced oxygen transport to tissue from the microcirculation. It can result in the death of cells and tissues, including entire organs as well as the patient. Blood clotting and sludging are common occurrences during and after invasive surgery; thus, it is imperative to find safe procedures to reduce or prevent these deadly phenomena. All anticoagulants used today, for clot prevention and dissolution, can cause excessive bleeding that can lead to enormous medical expense to provide control, otherwise causing patient death. Protein C is a natural protein and is the pivotal anticoagulant in the blood. Due to the mechanism of converting the zymogen protein C (ZPC) to active protein C (APC), only when and where it is needed, and their respective half-lives in the body, the natural anticoagulant, antithrombotic characteristics of APC can be utilized without causing bleeds.

Murine zymosan-induced peritonitis represents a well-defined model of acute inflammation. However, the molecular mechanisms by which leukocytes degrade basement membranes during extravasation into the peritoneum are not clear. Gelatinase B (MMP-9) is thought to participate in cellular migration, yet its role in leukocyte transmigration through endothelia during inflammation remains controversial. The aim of the present study was to evaluate the role of MMP-9 in the cell influx during zymosan-induced experimental peritonitis. In zymosan-treated Balb/c mice MMP-9 and its natural inhibitor (tissue inhibitor of metalloproteinase 1 - TIMP-1) were present in the peritoneal fluid and plasma at the time of peritoneal neutrophil (polymorphonuclear leukocyte - PMN) infiltration and persisted there until the time of monocytes/macrophages influx. To probe the function of gelatinases, gelatinase B-deficient mice (MMP-9(-/-)) were used as well as Balb/c mice treated with cyclic CTTHWGFTLC (INH), a specific peptide inhibitor of gelatinases. The studies revealed that in either group of mice deprived of MMP-9 activity, PMN infiltration was impaired at the time of their maximal extravasation (6h) while tumor necrosis factor alpha (TNF-alpha), cytokine-induced neutrophil chemoattractant (KC) and interleukin 10 (IL-10) levels were not changed. At later stages (24 h post-zymosan) a significant increase in PMNs was observed in MMP-9(-/-) mice, but not in the inhibitor-treated mice, in comparison to their respective controls. Moreover, intraperitoneal (i.p.) injection of recombinant mouse pro-MMP-9 induced leukocyte accumulation in peritoneum. Collectively, the findings indicate that gelatinase B participates in leukocyte transmigration; however, its function can be compensated by other mechanisms.

Hepatitis C virus (HCV) encodes a polyprotein consisting of core, envelope (E1, E2, p7), and nonstructural polypeptides (NS2, NS3, NS4A, NS4B, NS5A, NS5B). The serine protease (NS3/NS4A), helicase (NS3), and polymerase (NS5B) constitute valid targets for antiviral therapy. We engineered BH3 interacting domain death agonist (BID), an apoptosis-inducing molecule, to contain a specific cleavage site recognized by the NS3/NS4A protease. Cleavage of the BID precursor molecule by the viral protease activated downstream apoptotic molecules of the mitochondrial pathway and triggered cell death. We extended this concept to cells transfected with an infectious HCV genome, hepatocytes containing HCV replicons, a Sindbis virus model for HCV, and finally HCV-infected mice with chimeric human livers. Infected mice injected with an adenovirus vector expressing modified BID exhibited HCV-dependent apoptosis in the human liver xenograft and considerable declines in serum HCV titers.

The lung is exposed to high oxygen tension and oxygen free radicals have been implicated in many pathologies of the organ. Extracellular superoxide dismutase occurs in high concentration in the lung and protects against hyperoxia-induced inflammation. We hypothesized that the enzyme might ameliorate other types of inflammation as well as aging-related changes of the organ. Tracheal instillation of endotoxin plus zymosan into extracellular superoxide dismutase knockout and wild-type mice resulted in a marked neutrophilic inflammation and increases in inflammatory cytokines, protein, and lactate dehydrogenase activity in the bronchoalveolar lavage fluid. There were no significant differences between the genotypes. Repeated challenges with ovalbumin caused an allergic inflammation with increases in eosinophils, interleukin-5, protein, and lactate dehydrogenase activity in the bronchoalveolar lavage fluid. Only minimal differences between the genotypes were found. In lungs from 2-year-old mice, marginal increases in inflammatory variables and fibrosis were found in the knockout mice. In conclusion, extracellular superoxide dismutase had a negligible role in the present inflammation and allergy models and for the long-term integrity of the organ.

Background: The intraarterial administration of thrombolytic agents is associated with clinical benefits in patients with acute peripheral arterial occlusion, and urokinase has been the agent that has become the standard of care in the United States. Recombinant prourokinase (r-ProUK) offers potential as a novel agent with improved fibrin specificity and, as such, may offer advantages as an attractive alternative to urokinase., Methods: A randomized, double-blind, parallel, phase II, prospective multicenter trial was undertaken to compare three doses of intra-arterial, catheter-directed r-ProUK (2 mg, 4 mg, or 8 mg/hr for 8 hrs, then 0.5 mg/hr) versus one dose of tissue-culture urokinase (4,000 IU/min for 4 hrs, then 2,000 IU/min) for the treatment of acute lower extremity arterial occlusion of 14 days' duration or less (n = 241). The primary endpoint was complete (>95%) lysis of the occluding thrombus after 8 hours of infusion., Results: Increased clot lysis at 8 hours, decreased fibrinogen concentration, and an increased rate of hemorrhagic events were observed as the r-ProUK dose was increased from 2 mg/hr to 8 mg/hr. Similarly, a decreased duration of study drug infusion was seen, decreasing from 16.7 +/- 0.90 hours in the 2 mg/hr group to 12.7 +/- 0.97 hours in the 8 mg/hr group. The results for the urokinase group decreased to a level between those observed for the 2 mg and 8 mg r-ProUK group with respect to clot lysis at 8 hours, fibrinogen decrement, and bleeding complications, approximating those observed in the 4 mg/hr r-ProUK group. These results were achieved with a relatively low rate of major bleeding events and no episodes of intracranial hemorrhage., Conclusions: The 8 mg/hr dose of r-ProUK was associated with an increased rate of thrombolysis relative to the other treatment groups, associated with a slightly increased frequency of bleeding complications and decrements in fibrinogen concentration. Conversely, the 2 mg/hr r-ProUK dose was associated with a slightly slower rate of thrombolysis, but bleeding complications and fibrinogenolysis were diminished. r-ProUK is a novel thrombolytic agent with a dose-related safety and efficacy profile. As such, it offers potential as a useful tool in the treatment of peripheral vascular occlusion.

The physiological role of prorenin is unknown; however, the possibility that prorenin inhibits renin locally has been suggested. We tested the hypothesis that prorenin may be an endogenous competitor for renin uptake in the tissue. We also investigated whether prorenin can be activated to active renin and affect mean arterial pressure (MAP). Isolated perfused hindquarters of rats transgenic for human angiotensinogen were infused with human renin and/or prorenin. The plateau phase of angiotensin (Ang) I release 15 minutes after cessation of infusions was used as a parameter for renin uptake. Renin (10 ng/mL for 15 minutes) caused sustained release of Ang I (153+/-16 fmol/mL). Coinfusion with a 15-fold excess of prorenin did not affect local Ang I formation (153+/-19 fmol/mL). Prorenin infusion alone showed no activation to active renin. In addition, we investigated MAP and plasma Ang II levels after injection of saline (DeltaMAP, -1+/-2 mm Hg; 40+/-5 fmol/mL Ang II), 9 ng renin (DeltaMAP, +37+/-3 mm Hg; 378+/-39 fmol/mL), and 144 ng prorenin (DeltaMAP, +10+/-5 mm Hg; 61+/-5 fmol/mL) and the coinjection of renin and prorenin (DeltaMAP, +41+/-4 mm Hg; 305+/-23 fmol/mL) in anesthetized rats. The data show that prorenin was not activated to active renin and did not affect MAP in short-term experiments. Renin-induced Ang formation was not affected by prorenin. Renin may have been taken up specifically because of its physical and chemical properties or because of nonspecific sequestration in the extravascular space. We conclude that prorenin does not act as an endogenous antagonist for the long-lasting effects of renin in the vascular wall. Moreover, prorenin does not affect acute renin-related effects on blood pressure.

Background and Purpose: To test the safety and recanalization efficacy of intra-arterial local delivery of plasminogen activators in acute ischemic stroke, a randomized trial of recombinant pro-urokinase (rpro-UK) versus placebo was undertaken in patients with angiographically documented proximal middle cerebral artery occlusion., Methods: After exclusion of intracranial hemorrhage by CT scan, patients with abrupt onset of symptoms of focal ischemia likely to receive treatment within 6 hours who satisfied all clinical eligibility criteria underwent carotid angiography. Patients displaying Thrombolysis in Acute Myocardial Infarction grade 0 or 1 occlusion of the M1 or M2 middle cerebral artery were randomized 2:1 to receive rpro-UK (6 mg) or placebo over 120 minutes into the proximal thrombus face. All patients received intravenous heparin. Recanalization efficacy was assessed at the end of the 2-hour infusion, and intracerebral hemorrhage causing neurological deterioration was assessed at 24 hours., Results: Of 105 patients who underwent angiography, 59 were excluded from randomization. Among the 46 patients randomized, 40 were treated with rpro-UK (n=26) or placebo (n=14) a median of 5.5 hours from symptom onset. Recanalization was significantly associated with rpro-UK (2P=.017). Hemorrhagic transformation causing neurological deterioration within 24 hours of treatment occurred in 15.4% of the rpro-UK-treated patients and 7.1% of the placebo-treated patients (2P=.64). Both recanalization and hemorrhage frequencies were influenced by heparin dose., Conclusions: Intra-arterial local rpro-UK infusion was associated with superior recanalization in acute thrombotic/ thromboembolic stroke compared with placebo. In this regimen, heparin dose influenced hemorrhage frequency and recanalization. Although symptomatic hemorrhage remains a concern, this study suggests that recanalization is enhanced with rpro-UK and heparin.

Thrombolytic enzymes are widely used in the treatment of vascular diseases of the eyes and of intraocular hemorrhages. We studied the pharmacokinetics of recombinant prourokinase (proRUK) in ocular structures after its intravitreal administration and subtenon's implantation of collagen infusion system (SICIS). Kinetic parameters of accumulation of labeled proRUK in ocular structures were obtained. After intravitreal administration, the proRUK half-life (T1/2) was 7.9 +/- 1.44 h, with proRUK losing none of its enzymatic activity while in the vitreous body. The maximum accumulation of proRUK in the vitreous administered by the SICIS is observed after 4-5 h and is 1.5-2.0% of the dose administered. In the sclera the maximum accumulation of proRUK is 10-15% (4-6 h after administration), in the vascular membrane 0.8% (4 h after administration). We believe that combination of intravitreal and SICIS-aided administration of proRUK to patients with extensive hemorrhages (subtotal and total hemopthalmia) will improve the efficacy of therapy.

The past year has seen tremendous progress in developing new therapies aimed at reversing the effects of acute stroke. Thrombolytic therapy with various agents has been extensively studied in stroke patients for the past 7 years. Tissue plasminogen activator (t-PA) received formal US Food and Drug Administration approval in June 1996 for use in patients within 3 hours of onset of an ischemic stroke. Treatment with t-PA improves neurologic outcome and functional disability to such a degree that, for every 100 stroke patients treated with t-PA, an additional 11-13 will be normal or nearly normal 3 months after their stroke. The downside of t-PA therapy is a 6% rate of symptomatic intracerebral hemorrhage (ICH) and a 3% rate of fatal ICH. Studies are under way to determine whether t-PA can be administered with an acceptable margin of safety within 5 hours of stroke, to evaluate the therapeutic benefits of intraarterial pro-urokinase, and to assess the use of magnetic resonance spectroscopy to identify which patients are most likely to benefit from thrombolysis. Combination thrombolytic-neuroprotectant therapy is also being studied. In theory, patients could be given an initial dose of a neuroprotectant by paramedics and receive thrombolytic therapy in the hospital. We are now entering an era of proactive, not reactive, stroke therapies. These treatments may reverse some or all acute stroke symptoms and improve functional outcomes.

Peptides with sequences based on the leader sequence of yeast cytochrome c oxidase subunit IV (pCOX IV-(1-25)) activate the electrophoretic uptake of K+ and other cations such as tetraethylammonium and lysine by rat liver mitochondria with EC50 = 11-15 microM. Uptake of these cations is dependent on respiration and is prevented by uncoupling agents, and the Vmax for K+ is 1.2-1.5 micromol/min/mg. Albeit more slowly, the non-electrolytes mannitol and sucrose are also transported by this pathway. Treatment of the peptides with proteinase K eliminates the stimulatory effect. Since the stimulated rate is not inhibited by ATP or by cyclosporin, we conclude that this pathway is not related to the mitochondrial KATP channel or the Ca2+-dependent permeability transition pore. Transport is stimulated by pCOX IV-(1-23), pCOX IV-(1-22), and pCOX IV-(1-12)Y, but not by a 13-amino acid peptide representing the nuclear location sequence of the SV40 large T antigen, which is responsible for directing that protein to the nucleus. Spermine, which has four positive charges, also has no stimulatory effect, and an amphiphilic 22-residue peptide derived from antithrombin III with seven net charges is only one-twentieth as effective as pCOX IV-(1-22). Thus, these data indicate that the sequence/structure is important for activation of transport. We also demonstrate that mitochondrial uncoupling, previously reported to be induced by these peptides, actually reflects coupled accumulation of salt. In view of our findings, it is also likely that the lytic effects attributed to these peptides are secondary to swelling and are not due to membrane damage per se. Finally, we show that, in non-ionic media, the peptide is an inhibitor of cytochrome c oxidase.

A 69-year-old woman with combined valvular heart disease (mitral regurgitation and aortic regurgitation), ascending aortic aneurysm, and atrial fibrillation underwent double valve replacement (DVR) and, ascending aortic wall plication. The postoperative thrombo-test level was around 20%. The ST elevation on ECG (II, III, aVFm, V4 approximately V6) with chest pain were recognized on the 13 th postoperative day. She was diagnosed as having acute myocardial infarction, and percutaneous transluminal coronary recanalization was performed immediately. The coronary angiogram showed occlusion at the left anterior descending branch (#8). This lesion could be recanalized by 6,000 U plasminogen pro activator (pro-UK) administration. The cineangiogram on the 35th postoperative day, revealed complete recanalization of this occlusion. Several cases of acute myocardial infarction associated with valvular heart diseases has been reported previously in Japan. However, there has been no report, except for this case, demonstrating occlusion in the coronary artery after prosthetic replacement and successful PTCR. So, this case is the first report on that point.

Purpose: Despite advantages demonstrated in vitro, no single thrombolytic agent has been clearly shown to be superior to another in the clinical setting. Prourokinase has recently received attention as a new thrombolytic agent with higher fibrin specificity. The thrombolytic activity of prourokinase, however, remains ill defined. The purpose of this study was to evaluate thrombolysis with prourokinase in comparison to urokinase in vitro., Methods: We used an in vitro parallel channel perfusion model that simulates catheter-directed thrombolysis in the peripheral arterial system. Radiolabeled thrombi were subjected to 90 minutes of endhole catheter-directed infusion with either prourokinase 5000 IU/ml, urokinase 5000 IU/ml; or 5% dextrose in water at 4 ml/hr., Results: Prourokinase and urokinase were found to be equivalent with respect to thrombolytic effect. Percent lysis was maximal at 90 minutes in both the urokinase and prourokinase groups. Prourokinase and urokinase were found to be equally effective in restoring flow through thrombosed graft segments., Conclusion: Prourokinase appears to offer little benefit over urokinase with respect to thrombolytic activity in an in vitro model that closely resembles the clinical setting. If prourokinase is to be accepted as an alternative to urokinase, advantages must relate to differences in fibrin specificity.

Objectives: Three dosage regimens of a new recombinant glycosylated prourokinase (A-74187) were evaluated by measuring coronary artery patency at 90 min in patients with acute myocardial infarction., Background: Prourokinase is a thrombolytic drug with unique pharmacologic properties that may be clinically advantageous., Methods: Aspirin (325 mg), intravenous heparin and prourokinase (60- or 80-mg monotherapy or 60 mg "primed" with a preceding bolus dose of 250,000 IU of recombinant urokinase) were administered to 128 patients. Coronary angiography was performed at 60 min (wherever possible), 90 min (primary end point) and 24 h to determine arterial patency and reocclusion rates. Plasma was collected serially to measure fibrinogen, plasminogen, thrombin antithrombin III and fibrinopeptide A. Clinical events until hospital discharge were recorded., Results: The coronary artery patency rate at 90 min was similar for all three regimens, averaging 73% (95% confidence interval [CI] 64% to 80%); Thrombolysis in Myocardial Infarction (TIMI) grade 3 flow rates averaged 52% (95% CI 42% to 61%). Arterial patency at 60 min was 62% (95% CI 50% to 73%), and reocclusion occurred in 1.4% (95% CI 0.1% to 4.1%). Prourokinase demonstrated relative fibrin specificity at all doses studied. Fibrinopeptide A and thrombin antithrombin III levels were elevated at baseline and declined rapidly during the 1st 12 h. There was no difference in the baseline values of these thrombin markers between patients with patent versus closed arteries at 90 min. There was one death; no strokes occurred., Conclusions: A-74187 prourokinase is a rapid-acting, effective fibrin-specific thrombolytic agent. Reocclusion was unusual, possibly because of aggressive anticoagulation with intravenous heparin or unique features of the drug. Full definition of the clinical effectiveness of this drug merits examination in future randomized trials evaluating clinical and angiographic effectiveness.

The effects of thrombolytic treatment was studied in 109 consecutive patients 9-11 days after their first acute myocardial infarction by high-resolution electrocardiography (ECG), 24 h Holter monitoring, exercise test and radionuclide ventriculography. Thirty-seven patients were treated with intravenous thrombolytic agents. Thrombolytic treatment was assessed by clinical criteria to be successful in 22 patients and probably successful in 12 patients. Thrombolysis failed in three patients and 72 patients did not receive thrombolytic treatment (control group). Measurements made on the high-resolution and filtered (60 Hz high-pass) vectormagnitude complex included the total duration, the duration of the potential less than 40 microV, the root mean square (RMS) voltage in 10 ms intervals over the first 50 ms and RMS voltage of the last 40, 50 and 60 ms. The filtered QRS duration was significantly shorter in reperfused patients compared with the control group (83 +/- 10 vs 89 +/- 12 ms; P = 0.017). In inferior infarcts (n = 57) the filtered QRS duration was 83 +/- 11 ms in reperfused and 89 +/- 10 ms in non-reperfused patients (P = 0.044), but in anterior infarcts (n = 52) there was no difference. The RMS voltage of the initial 50 ms of the QRS was higher in the reperfused than in non-reperfused anteroseptal infarcts (38 +/- 14 v 23 +/- 10 microV; P = 0.022). Patients successfully treated with thrombolytic agents within the first 2 h had higher RMS voltage of the terminal 40 ms of the QRS than patients treated within 2-4 h (38 +/- 17 vs 27 +/- 17 microV; P = 0.03).(ABSTRACT TRUNCATED AT 250 WORDS)

In a multicenter dose-finding study, the thrombolytic potency of urokinase preactivated pro-urokinase was evaluated. Sixty-two patients were randomly assigned to receive 250,000 U of urokinase plus either 4.5 mega U (group I: n = 33) or 6.5 mega U (group II: n = 29) of pro-urokinase. Patency rates were 36.4% (20.4-54.9%) vs. 54.5% (36.3-71.9%) (n = 27) at 60 minutes and 55.6% (32.5-70.6%) vs. 62.1% (42.3-79.3%) at 90 min into thrombolysis (n.s.). In a third group of 12 patients treated with 500,000 U of urokinase plus 6.5 mega U of pro-urokinase patency was achieved in 33.3% (9.9-65.1%) and 41.7% (15.2-72.3%) at 60 and 90 min, respectively. Patency rates at 24 hr follow-up angiography (n = 35) were 78.6% (49.2-95.3%), 85.7% (57.2-98.2%), and 85.7% (42.1-99.6%). Coagulation analysis in 37 patients revealed similar alterations in the three treatment groups with minor decreases in fibrinogen levels, moderate drops in plasminogen and alpha-2-antiplasmin levels, and moderate increases in the concentrations of the total fibrinogen/fibrin degradation products, the differences between the groups not being significant. Bleeding complications were observed in 12.9%, 13.8%, and 25% of patients in groups I, II, and III, respectively, mainly related to catheter sites. Hence, the safety profile of urokinase preactivated pro-urokinase seems comparable to other thrombolytic regimens. Reopening of occluded coronary arteries, however, is achieved relatively slowly. Thus, in its use for thrombolysis in myocardial infarction, urokinase preactivated pro-urokinase does not seem to offer superior advantages.

Enhancement by anticoagulation of thrombolysis with infused or bolus-injected saruplase (r-scu-PA) has been studied using heparin and the thrombin inhibitor argatroban. In a rabbit femoral artery thrombosis model infusion of saruplase (3 - 12 mg/kg, 60 min) caused a dose-dependent thrombolysis. Reperfusion rate after infusion of 3 mg/kg saruplase alone was 3/6, reperfusion time 42 +/- 3 min and reocclusion rate 2/3; final patency rate at 120 min was 17%. Combination of 3 mg/kg saruplase with heparin (150 U/kg + 100 U/kg.hr i.v.; 5.3-fold PTT-prolongation) resulted in a reperfusion rate of 6/6 after a reperfusion time of 39 +/- 7 min; reocclusion rate was 3/6 and final patency rate was 50%. Argatroban (1 mg/kg + 3 mg/kg.hr i.v.; 2.3-fold PTT prolongation) in combination with saruplase resulted in a reperfusion rate of 6/6 after 26 +/- 5 min; no reocclusion occured and final patency rate was 100% (p less than 0.05 vs saruplase alone). Bolus injection of 6 mg/kg saruplase achieved reperfusion in 5/6 arteries after 15 +/- 3 min, but reocclusion rate was 4/5; final patency rate was 17%. Combination of bolus-injected saruplase with heparin resulted in a reperfusion rate of 4/6 after 8 +/- 3 min and no reocclusion occured; patency rate was 67%. With combination of argatroban and bolus-injected saruplase 6/6 arteries were reperfused after 8 +/- 3 min; reocclusion was prevented and final patency rate was 100% (p less than 0.05 vs saruplase-bolus alone). Systemic fibrinogenolysis was more pronounced with bolus injection than infusion of saruplase. The results indicate that arterial thrombolysis with saruplase can be enhanced by heparin and the thrombin inhibitor argatroban. The bolus injection of saruplase resulted in persistent reperfusion when simultaneous anticoagulation was performed. Despite less PTT prolongation, enhancement of saruplase-induced thrombolysis was more effective with argatroban than with heparin in rabbit femoral artery thrombosis.

When in the experiment on rats the jointly immobilized on a carbonic cloth proteases and lytic enzyme complex were used, the reduced period of the suppurative wound cleaning and healing was noted as compared to that in use of the immobilized on a carbonic cloth proteases and jointly immobilized on a gause proteases and lytic enzyme complex.

Coronary thrombolysis with plasminogen proactivator, GE-0943, was undertaken in patients with acute myocardial infarction and compared with urokinase. Fifty patients were given 6000 units of GE-0943 intracoronary and the totally occluded infarct-related arteries were recanalized in 92.0%. Fifty-four patients who were given 3000 units of GE-0943 intracoronary demonstrated a recanalization rate of 68.2%, while intracoronary administration of urokinase in 54 patients showed a recanalization rate of 70.4%. There was only one patient with hemorrhagic complication among GE-0943 3000-units group, none among GE-0943 6000-units group, and 5 among urokinase group. Thus, intracoronary administration of GE-0943 is very effective and safe with a dose up to 6000 units.

A precursor form of cathepsin B (Mr 45-47 kd) was purified from ascitic fluids of patients with ovarian adenocarcinomas. Following pepsin activation, this precursor produced a 33 kd cathepsin B-like proteinase closely related to lysosomal cathepsin B. A similar activation was found using the 52 kd pro-cathepsin D secreted by the MCF7 human breast cancer cells. This activation was a time, dose and pH dependent process. These results suggest that the 52 kd pro-cathepsin D may be involved in the early steps of the "metastatic cascade", activating pro-cathepsin B in an acidic environment.