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PubMed1987

Migration of neutrophils across monolayers of cultured microvascular endothelial cells. An in vitro model of leucocyte extravasation.

Monolayers of bovine microvascular endothelial cells (BMECs) grown on connective tissue derived from human amniotic membrane were used to examine the transendothelial migration of human neutrophils in vitro. Neutrophils placed above these cultures migrated in response to a chemotactic gradient generated by placing 10(-7) M-formyl-methionyl-leucyl-phenyl-alanine (fMLP) below the cultures. Under these conditions, an average of 29 +/- 12% of the total population of neutrophils migrated beneath the endothelium after 1 or 2 h of incubation. Neutrophil migration in the absence of fMLP or in the presence of equal concentrations of fMLP above and below the cultures was less than 8% of the response to a 10(-7) M-fMLP gradient. Migration was a rapid event. Neutrophils began adhering to the apical surface of the endothelium within 2 min following exposure to an fMLP gradient; Ca2+ was required for this initial adhesion. Within 10 min, the majority of neutrophils associated with the BMEC-amnion cultures had migrated beneath the endothelial monolayer. Ultrastructural studies revealed that the initial adhesion between migrating neutrophils and endothelium was characterized by close contact between the two types of cell in focal areas. This close association was maintained as the neutrophils traversed the clefts between endothelial cells. Following their migration across the endothelium, neutrophils often were observed lying between the endothelium and its basement membrane. With time, the neutrophils penetrated the basement membrane and moved into the underlying amniotic connective tissue. To test the role of neutrophil proteinases in breaching endothelial and subendothelial barriers, migration was allowed to proceed in the presence of a variety of proteinase inhibitors, including p-nitrophenyl p'-guanidinobenzoate, soybean trypsin inhibitor, 6-aminocaproic acid, alpha 1-proteinase inhibitor, leupeptin, antipain and methoxysuccinyl alanine-alanine-proline-valine chloromethyl ketone. None of these had a significant effect on the number of neutrophils that migrated or the depth to which they penetrated the amniotic tissue as compared with controls. In contrast, pepstatin and chymostatin reduced migration in response to fMLP to 7% and 52% of control values, respectively. However, these two inhibitors did not affect migration in response to another chemoattractant, leukotriene B4. Migration was neither enhanced nor inhibited by the following treatments: (1) removal of plasminogen from the calf serum used in the assay medium and addition of polyclonal antibody to plasminogen; (2) addition of monoclonal or polyclonal antibody to plasminogen activator.(ABSTRACT TRUNCATED AT 400 WORDS)

Calcium↗
PubMed1991

Purification and some properties of inducible lysine decarboxylase from Vibrio parahaemolyticus.

Inducible lysine decarboxylase from Vibrio parahaemolyticus AQ 3627 was purified to apparent homogeneity and characterized. The enzyme displayed a molecular weight of 531000 by gel filtration and 79000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The enzyme required pyridoxal phosphate as a cofactor, and the pH optimum was 5.5. The Km value for L-lysine was 3.2 mM, and the enzyme was inhibited by 6-aminocaproic acid and alpha-fluoromethyl analogs of cadaverine. delta-Hydroxylysine and S-aminoethyl-L-cysteine was active as substrates to a lesser extent than L-lysine. The amino-terminal amino acid sequence was determined to be Met-Asn-Ile-Phe-Ala-Ile-Leu. These properties were compared with those of other bacterial lysine decarboxylases.

Amino Acid Sequence↗
PubMed1985

Inability of rat anaphylatoxin to induce histamine release in rats.

The role of rat anaphylatoxin in histamine release and increased vascular permeability during the first thirty minute period in zymosan-air-pouch inflammation, an experimental model of inflammation induced by zymosan in an air-pouch prepared on the back of rats, was investigated. Complement depletion by cobra venom factor did not affect the histamine release nor the increased vascular permeability in the inflammation of this type. In spite of apparent anaphylatoxin activity, zymosan activated serum (ZAS) failed to cause any significant release of histamine when infused in the air-pouch on the back. Anaphylatoxin purified from rat serum activated with zymosan in the presence of an inhibitor (epsilon-aminocaproic acid) of anaphylatoxin inactivator gave a single band in both polyacrylamide gel electrophoresis (PAGE) and SDS-PAGE. The molecular weight estimated by SDS-PAGE was approx. 7,000. The purified rat anaphylatoxin failed to induce histamine release nor increased vascular permeability even at 50 micrograms/ml, although it caused contraction of guinea pig ileum at 0.8 micrograms/ml. These results suggest that rat anaphylatoxin does not participate in histamine release and increased vascular permeability in the zymosan-air-pouch inflammation.

Anaphylatoxins↗
PubMed2006

Comparison of capillary zone electrophoresis and isotachophoresis determination of dimethindene enantiomers in pharmaceuticals using charged carboxyethyl-beta-cyclodextrin as a chiral selector.

Charged cyclodextrin-mediated capillary electrophoretic methods (capillary zone electrophoresis [CZE] and isotachophoresis [ITP]) in a hydrodynamically closed separation system with suppressed electroosmotic flow and UV absorbance photometric detection (for CZE) or conductivity detection (for ITP) were developed for a highly effective separation and quantitation of dimethindene enantiomers in various pharmaceutical formulations (solution, gel, capsules). Optimized electrolyte systems were based on low-mobility buffer constituents (epsilon-aminocaproic acid, beta-alanine, potassium acetate, acetic acid), negatively charged (at pH > 4.5) carboxyethyl-beta-cyclodextrin (CE-beta-CD), serving as chiral selector, and methylhydroxyethylcellulose, serving as an electroosmotic flow suppressor. Complete enantioseparations of dimethindene in the presence of low concentrations of CE-beta-CD (2.5 mg/ml in CZE, 6.0 mg/ml in ITP) clearly indicated a role of charge of this chiral selector in enantioresolution. It also outlined the potential of charged CD-derivatives as chiral selectors for various CE techniques. The proposed methods were successfully validated, appraising parameters of sensitivity, linearity, precision, accuracy/recovery, and robustness, and then, they were applied to pharmaceutical samples. Consistent results obtained by both CE methods indicate their usefulness for routine use.

Dimethindene↗
PubMed2004

Expansion in intracerebral hematoma.

Intracerebral hemorrhage (ICH) is the last stroke subtype without a primary therapy. The major research question is whether to treat the whole chain of disease events or just stabilize one link in the process. Several candidate events that would benefit from treatment exist: hemorrhage extension, mass effect-clot removal, or delayed edema-inflammation. In this issue of Neurocritical Care, Piriyawat et al. present a pilot study using two convenience samples of ICH patients exploring the idea that blood clot stabilization with antifibrinolytic drugs would limit the early expansion of hematomas. Two groups of patients were studied in sequence: nine ICH patients who experienced two episodes of hematoma expansion (HE) served as controls, demonstrating a baseline hematoma extension event rate of 22%. A second group of five ICH patients was treated in the first 12 hours after symptoms with the antifibrinolytic drug e-aminocaproic acid. Three of the treated patients experienced HE for an event rate of 60%. On first pass, it seems that clot stabilization is not likely to occur with eanimocaproic acid. The author's institutional IRB concluded that treatment in this protocol was futile and requested the study be stopped.

Antifibrinolytic Agents↗
PubMed1985

Dental management of a patient with glycogen storage disease type I.

Patients with glycogen storage disease type I (GSD I) have varied medical problems, including a bleeding diathesis characterized by a prolonged bleeding time. A case of a patient with GSD I having a history of medical problems, some of which were related to oral and dental bleeding, was presented. The patient was cariesfree and had generalized moderate to severe periodontal breakdown. A treatment regimen in a hospital setting, using cryoprecipitate and aminocaproic acid, permitted dental care and control of oral bleeding and associated complications.

Adult↗
PubMedDate not supplied

Comparison of an indirect format ELISA on modified graphite and polystyrene surfaces against triazines.

An indirect format enzyme-linked immuno-sorbent assay (ELISA) on graphite rods ( 0.8 mm x 20 mm) and, for comparison, on microtiter plates has been developed against terbuthylazine. For this purpose, a series of 2-aminoalkyl-4-chloro-6-terbuthyl-s-triazine-2,6-diamine ELISA haptens with alkyl spacer lengths of 2, 4, 6, and 8 CH2 groups has been synthesized. The graphite rods or the microtiter plates were covered by a polymerized glutaraldehyde network, and the ELISA haptens have been coupled by imino coupling to the free aldehyde groups of that network. epsilon-Aminocaproic acid has been used as an agent to block unspecific binding sites. The ELISA was run in a competitive mode, where the immobilized ELISA hapten and the solute terbuthylazine as a target analyte compete for the solute antibody.

Enzyme-Linked Immunosorbent Assay↗
PubMed1999

Differential expression of proteases in human gestational tissues before, during and after spontaneous-onset labour at term.

A number of tightly regulated proteolytic enzyme systems, including the plasminogen activation cascade and matrix metalloproteases, play integral roles in the remodelling of extracellular matrices during pregnancy and parturition. This study assessed these labour-associated changes in protease activity in human gestational tissues. Amnion, choriodecidua and placenta collected from women before (at caesarean section, not in labour), during (at caesarean section, in labour) and after (spontaneous-onset labour, normal vaginal delivery) labour were examined on gelatin-substrate SDS-PAGE zymography. All tissues displayed major 55 kDa plasminogen-dependent activity that was abolished by the serine protease inhibitors (10 mmol phenylmethyl-sulphonylfluoride l-1, 100 mmol epsilon aminocaproic acid l-1, 1 mmol Glu-Gly-Arg chloromethylketone l-1). The enzymic activity was identified as urokinase plasminogen activator on the basis of its co-migration with reference standard and western blot analysis, and did not vary with labour status. An additional protease with an apparent molecular mass of approximately 90 kDa was detected in all tissues. Densitometric measurement of these tissues showed a significant (P < 0.05) increase in this enzyme activity with labour onset. Heavy metal chelators (1 mmol 1.10 phenanthroline l-1 and 10 mmol EDTA l-1) selectively blocked the 90 kDa activity, consistent with the proposal that it is a metalloprotease. Co-migration with reference standard and western blot analysis confirmed the identity of this protease as the matrix metalloprotease 9 (MMP-9). Immunoreactive MMP-9 protein was also significantly (P < 0.05) increased during and after labour compared with before labour in all tissues examined. It is proposed that the upregulated expression of MMP-9 is involved in fetal membrane rupture and placental separation during and after labour onset, respectively. In conclusion, the regulated repertoire of protease activities expressed by human gestational tissues implies an important role for matrix-degrading enzymes during human parturition.

Amnion↗
PubMed1998

A risk-benefit assessment of aprotinin in cardiac surgical procedures.

Aprotinin, a naturally occurring serine protease inhibitor, has found widespread application during cardiac surgical procedures as a consequence of its ability to decrease blood loss and transfusion requirements. While its efficacy in a variety of clinical situations associated with increased risk of blood loss has been well established, at the same time, various complications including anaphylaxis, renal insufficiency, graft closure and arterial thromboses have been reported in association with aprotinin administration. In order to more fully evaluate the risks and benefits associated with aprotinin usage, this review first of all examines the hazards associated with transfusion of blood and blood products. Consideration is then given to various alternatives to allogeneic transfusion, including autologous predonation, acute normovolemic hemodilution, perioperative cell salvage and intraoperative plasma sequestration. A critique of other available pharmacological therapies, specifically desmopressin, aminocaproic acid and tranexamic acid, reviewing their modes of action, efficacy and associated complications, is then made. The role of aprotinin in cardiac surgery is then discussed and its pharmacology, including consideration of its antifibrinolytic, platelet preserving and anti-inflammatory effects is reviewed. Finally, an analysis of potential complications associated with aprotinin administration is undertaken. Issues involving its influence on specific measures of anticoagulation, namely partial thromboplastin time and activated clotting time, and issues relating to graft patency, hypothermic circulatory arrest, renal function, and allergic reactions are analysed and interpreted. In summary, this review concludes that most of the risks associated with aprotinin administration primarily involve inadequate anticoagulation and those of developing an allergic reaction, particularly upon aproptinin re-exposure. The benefits of aproptinin to decrease blood loss and transfusion requirements are confirmed, and there is evidence pointing to the intriguing possibility of a potential salutary effect on perioperative central nervous system complications.

Animals↗
PubMed1982

Adverse effects of drugs on muscle.

A variety of drugs used in clinical practice may cause myopathy or interfere with neuromuscular transmission. The precise incidence of such disorders is not known, but it is almost certainly higher than is generally suspected. An important aspect of drug-induced muscular disorders is their reversibility if the offending agent is withdrawn, whereas failure to do so may lead to unnecessary morbidity. The study of drug effects on muscle provides a means of investigating the pathological reactions of muscle, and of producing experimental models of naturally occurring myopathies. Drug-induced myopathies may result from a direct toxic effect, which may be local when the drug is injected into a muscle or more diffuse when the drug is taken systemically, or may be secondary to electrolyte disturbances, muscle compression, ischaemia, neural activation or to the development of an immunological reaction directed against muscle. Repeated injections of antibiotics or drugs of addiction may lead to severe muscle fibrosis and contractures. A variety of drugs may cause an acute or subacute painful necrotising myopathy which may be associated with myoglobinuria, at times leading to acute renal failure. Clofibrate and epsilon aminocaproic acid are the drugs most frequently implicated, but a similar syndrome may occur in alcoholics and heroin addicts. Certain hypocholesterolaemic agents may induce myotonia by altering the sterol composition of the muscle cell membrane, while certain drugs including beta-adrenergic blockers and agonists, succinylcholine and diuretics may exacerbate or unmask pre-existing myotonia. In the syndrome of malignant hyperpyrexia, halothane, succinylcholine and various other agents may induce a potentially fatal state of muscular rigidity and hypermetabolism in susceptible individuals as a result of a defect in the calcium transport function of the sarcoplasmic reticulum and possibly of other cellular membranes. In corticosteroid myopathy, which is the most common form of drug-induced myopathy, there is selective atrophy of type 2 muscle fibres and the primary metabolic effect is an inhibition of RNA and protein synthesis, although protein degradation is also increased. Chloroquine and a number of related drugs with amphiphilic cationic properties may induce lysosomal storage myopathy, which may be associated with cardiomyopathy and with a more widespread form of lipidosis.

Abnormalities, Drug-Induced↗
PubMed1985

Production of the modified form of human plasminogen by alpha 2-macroglobulin-plasmin complexes.

It has been speculated that the modified form of plasminogen, a precursor of proteolytic enzyme plasmin in plasma, plays an important role in fibrinolysis in the blood. The present study was undertaken to examine the production by alpha 2-macroglobulin-plasmin complexes. alpha 2-Macroglobulin-plasmin complexes were purified from urokinase-activated plasma by affinity chromatography on lysine-Sepharose and gel filtration on Ultrogel AcA 22. The plasmin complex converted native plasminogen into the modified form more easily in the presence of epsilon-aminocaproic acid. The modification of native plasminogen by alpha 2-macroglobulin-bound plasmin was completely inhibited by aprotinin, and partly by soybean trypsin inhibitor. alpha 2-macroglobulin-bound plasmin produced modified plasminogen in human plasma where potent plasmin inhibitors exist, though the degree of production was small. The present results support the speculation of the important role of the modified form in vivo.

Animals↗
PubMed1994

Suppression of pentosidine formation in galactosemic rat lens by an inhibitor of aldose reductase.

Recent work from our laboratory revealed a correlation between the degree of protein pigmentation in human cataractous lens and the advanced Maillard reaction as reflected by pentosidine formation. Although the data suggested a role for ascorbate in pentosidine formation in senile cataractous lenses, elevated pentosidine levels in diabetic cataracts suggested that glucosylation may be involved directly in pentosidine biosynthesis. To clarify this issue, we quantified pentosidine in lenses from rats with experimental galactosemia with and without aldose reductase inhibitor treatment. At 12 months, pentosidine-like fluorescence (335/385 nm) was three to six times higher (P < 0.0001) in water soluble and insoluble crystallins of galactosemic compared with nongalactosemic rats. Actual pentosidine levels increased shortly after onset of galactosemia. Contents in water-insoluble crystallins were 6.32 +/- 2.2 and 1.40 +/- 0.66 pmol/mg protein in galactosemic and control lenses, respectively (P < 0.001). Fluorescence and pentosidine were suppressed to almost control levels upon treatment with sorbinil. Incubation experiments showed that pentosidine could form slowly from galactose, but much more rapidly from ascorbate and its oxidation products. Its formation could be inhibited partly by both reduced and oxidized glutathione or epsilon-aminocaproic acid. The requirement of oxygen for pentosidine formation suggests that oxidative stress associated with glutathione depletion and ascorbate oxidation are plausible mechanisms for rapid pentosidine formation upon onset of galactosemia. In contrast, Maillard reaction by glycoxidation products may account for the sustained increase in pentosidine. Both these events may be linked to the newly recognized pseudohypoxic state of cells exposed to high sugar concentrations.

Aldehyde Reductase↗
PubMed1976

A controlled investigation of the value of anticoagulant therapy in cases of prostatectomy.

Prior to prostatectomy, 182 patients were classified into two groups of 90 and 92 patients. The classification was performed so as to make the groups mutually comparable with respect to factors considered significant in connection with the risk of thrombo-embolism. The group with 90 patients was anticoagulated with marcoumar. The other group served as a control. There was no significant difference in mortality or thrombosis incidence between the two groups. There was a significantly larger loss of blood in the anticoagulated group. Five of the patients in the anticoagulated group died, while in the control group, 3 patients died. Epsilon-aminocaproic acid was not used on a routine basis.

Aged↗
PubMedDate not supplied

Successful multimodal therapy for kaposiform hemangioendothelioma complicated by Kasabach-Merritt phenomenon: case report and review of the literature.

We present the management challenge provided by a patient with kaposiform hemangioendothelioma associated with Kasabach-Merritt phenomenon. A female child presented at 14 months of age with an ecchymotic swelling of her right upper arm and axilla. Subsequently, she developed profound thrombocytopenia and hypofibrinogenemia (Kasabach-Merritt phenomenon). Biopsy of the lesion revealed kaposiform hemangioendothelioma, which has been reported as the predominant pathologic diagnosis associated with Kasabach-Merritt phenomenon. To achieve involution of the lesion and preserve function of the arm, the following interventions were involved: embolization, systemic interferon, cyclophosphamide, epsilon aminocaproic acid, and compression therapy. The clinical management of this patient was formidable until we arrived at the proper combination of therapies. Multimodal intervention may be required to manage fastidious hemangioendotheliomas of childhood, achieve clinical improvement, and prevent further morbidity.

Female↗
PubMed1979

Catheptic carboxypeptidase B as a major component in "T-cell activating factor" of macrophages.

TAF (T-cell activating factor), produced by peritoneal macrophages and assayed by its ability to potentiate DNA synthesis in macrophage-depleted lymph node cells stimulated with Phytochemagglutinin-concanavalin A, was shown to contain catheptic carboxypeptidase B (CPB) which accounted almost quantitatively for its potentiating activity. TAF action was inhibited by CPB inhibitors and could be mimicked by commercial pig pancreas CPB. The activity was absorbed from active supernatants on EACA-Sepharose and could be eluted with free EACA (epsilon aminocaproic acid).

Animals↗
PubMed1982

Results and complications of surgical management of 809 intracranial aneurysms in 722 cases. Related and unrelated to grade of patient, type of aneurysm, and timing of surgery.

Data from 722 consecutive causes with intracranial aneurysms were stored in a computer and later retrieved for analysis. Results and complications (including preoperative death and morbidity) of the surgical management of these patients were correlated with the Botterell grade of the patient in individuals with a recent subarachnoid hemorrhage (SAH), with the type of aneurysm, and with the timing of the surgical procedure. Patients with no SAH within 30 days prior to hospital admission were classified as "no SAH." Approximately 30% of all patients had sustained more than one hemorrhage. Death and morbidity rates prior to surgery in good-grade patients with a recent SAH exceeded the risk of surgery itself. Rebleeding was the primary cause for death and morbidity in Grade 1 patients: 3% of Grade 1 patients died from a recurrent hemorrhage and 7% deteriorated to a lower grade. Deterioration from ischemia produced by vasospasm related or unrelated to rebleeding exceeded the risks of rebleeding in Grade 2 patients. There was an operative morbidity of 2% and mortality of 2% in patients who were classified as Grade 1 at the time of surgery, but an overall management morbidity of 3% and mortality of 6% in patients who were in Grade 1 at the time of hospital admission. Early surgery in Grade 1 patients was not associated with an increased incidence of delayed ischemia postoperatively. In Grade 2 patients, the operative morbidity and mortality was 7% and 4%, respectively, and the management morbidity and mortality 16% and 11%, respectively. Early surgery in this group was associated with a high frequency of postoperative delayed ischemia (particularly in patients with more than one SAH). Epsilon-aminocaproic acid appeared to protect against a rebleed, gut was associated with a higher incidence of postoperative pulmonary emboli. Intraoperative complications were related both to the size of the aneurysm and to its location. Repair of multiple aneurysms did not adversely affect the result. The surgical approach, the importance of using a self-retaining brain retractor, and the technical complications in these cases are discussed.

Brain Ischemia↗
PubMed2006

Sacral epithelioid angiosarcoma associated with a bleeding diathesis and spinal epidural hematoma: case report.

Epithelioid angiosarcoma of bone is a rare, high-grade lesion that is highly vascular and can be associated with a bleeding diathesis. An association has been reported in angiosarcomas in other locations with coagulopathy from tumor-related disseminated intravascular coagulopathy and fibrinolysis. The authors report the case of a rare occurrence of a primary sacral epithelioid angiosarcoma associated with a large epidural hematoma and a severe bleeding diathesis. A 25-year-old woman presented with weakness, fatigue, neck and low-back pain, and progressive left S-1 radiculopathy. Imaging studies revealed a large ventral epidural hematoma extending from the sacral region rostrally to C-2 and a vascular tumor located in the sacrum. The patient underwent a sacral laminectomy, complicated by postoperative bleeding from the wound, and required massive transfusions. Ultimately, multimodal therapy was required to obtain hemostasis, including the use of endovascular embolization, radiation therapy, and an infusion of epsilon-aminocaproic acid with heparin. This case represents the first report of a primary epithelioid angiosarcoma in the sacrum and emphasizes that the coagulopathy seen in angiosarcoma is also a feature of this epithelioid variant.

Adult↗
PubMed1987

Haemophilus influenzae type b polysaccharide-protein conjugate vaccine.

An Haemophilus influenzae type b capsular polysaccharide-protein conjugate has been prepared. The polysaccharide was coupled to the serotype II protein of group B meningococcus through the spacer 6-aminocaproic acid using cyanogen bromide and water soluble carbodiimide. The conjugate can be shown to be reproducible and is stable and highly immunogenic in mice and African green monkeys. Clinical evaluation of this conjugate in children 3 months to 4 years of age showed that it elicited an antibody titer to the polysaccharide moiety greater than 1000 ng/ml in children 8 months of age or older.

Animals↗