Enzymes In Therapy Research Articles

Hemostatically Active Proteinase Produced by Aspergillus ochraceus: Key Specific Properties and Effect on Target Proteins.

The effect of A. ochraceus proteinase on the proteins of the human hemostasis system, fibrin, fibrinogen, plasminogen, protein C, and factor X, was studied. These proteins are key targets for proteolytic enzymes in therapy and diagnosis of thromboembolic complications. It was shown that A. ochraceus proteinase efficiently cleaves fibrin and fibrinogen, but does not act precisely, since it cuts all three subunits of these proteins. The proteinase did not have an activating effect on the plasminogen, a precursor of plasminogen and plasmin. The proteinase of A. ochraceus was shown to be the first fungal proteinase with proven activating activity towards the human hemostasis system factors protein C and factor X. For protein C activation, A. ochraceus proteinase requires Ca2+ ions. The enzyme was found to be sensitive to thrombin inhibitors, but not to plasmin inhibitors. A proteolytic action profile of the scope of this proteinase as a proteinase with activating protein C, factor X, and plasmin-like activity was proposed.

May Chronic Diseases be Treated by Intracellular Enzymes? An Opinion

More and more diseases do not find a basis at the level of causes. One of the causes is disturbances in the effectiveness of intracellular enzymes. Without their perfect function there is no life. Until now, no industrial production of these enzymes for therapy existed. This changed, because the company Citozeatec in Milano produces several enzyme preparations, which are active in the cells, mitochondria and cell nuclei. With these, success can be achieved in many chronic diseases.

Prussian Blue Nanozymes with Enhanced Catalytic Activity: Size Tuning and Application in ELISA-like Immunoassay.

Prussian blue nanozymes possessing peroxidase-like activity gather significant attention as alternatives to natural enzymes in therapy, biosensing, and environmental remediation. Recently, Prussian blue nanoparticles with enhanced catalytic activity prepared by reduction of FeCl3/K3[Fe(CN)6] mixture have been reported. These nanoparticles were denoted as ‘artificial peroxidase’ nanozymes. Our study provides insights into the process of their synthesis. We studied how the size of nanozymes and synthesis yield can be controlled via adjustment of the synthesis conditions. Based on these results, we developed a reproducible and scalable method for the preparation of ‘artificial peroxidase’ with tunable sizes and enhanced catalytic activity. Nanozymes modified with gelatin shell and functionalized with affine molecules were applied as labels in colorimetric immunoassays of prostate-specific antigen and tetanus antibodies, enabling detection of these analytes in the range of clinically relevant concentrations. Protein coating provides excellent colloidal stability of nanozymes in physiological conditions and stability upon long-term storage.

Adenosine-Metabolizing Enzymes, Adenosine Kinase and Adenosine Deaminase, in Cancer.

The immunosuppressive effect of adenosine in the microenvironment of a tumor is well established. Presently, researchers are developing approaches in immune therapy that target inhibition of adenosine or its signaling such as CD39 or CD73 inhibiting antibodies or adenosine A2A receptor antagonists. However, numerous enzymatic pathways that control ATP-adenosine balance, as well as understudied intracellular adenosine regulation, can prevent successful immunotherapy. This review contains the latest data on two adenosine-lowering enzymes: adenosine kinase (ADK) and adenosine deaminase (ADA). ADK deletes adenosine by its phosphorylation into 5′-adenosine monophosphate. Recent studies have revealed an association between a long nuclear ADK isoform and an increase in global DNA methylation, which explains epigenetic receptor-independent role of adenosine. ADA regulates the level of adenosine by converting it to inosine. The changes in the activity of ADA are detected in patients with various cancer types. The article focuses on the biological significance of these enzymes and their roles in the development of cancer. Perspectives of future studies on these enzymes in therapy for cancer are discussed.

Hydrolytic enzymes in therapy of gestosis

The clinical effectiveness o f complex enzymotherapy with preparation Vobenzim in treatment o f gestosis was studied in comparison with traditional therapy. The author analyzes the course оf gestation and delivery, and the state оf neonates in women with nephropathy o f I degree, who got common treatment (28 patients) and complex treatment including Vobenzim (17patients). Positive clinical effect o f complex enzymotherapy in case o f nephropathy is displayed in decreasing of edematic syndrome, eliminating or decreasing o f proteinuria, lowering o f blood pressure and impairment o f fetalhypoxia in labors. It goes along with the reduction o f immune complexes fixed in placenta.

Enzymes in therapy of biofilm-related oral diseases.

Biofilm-related infections of the oral cavity, including dental caries and periodontitis, represent the most prevalent health problems. For years, the treatment thereof was largely based on antibacterial chemical agents. Recently, however, there has been growing interest in the application of more preventive and minimally invasive biotechnological methods. This review focuses on the potential applications of enzymes in the treatment and prevention of oral diseases. Dental plaque is a microbial community that develops on the tooth surface, embedded in a matrix of extracellular polymeric substances of bacterial and host origin. Both cariogenic microorganisms and the key components of oral biofilm matrix may be the targets of the enzymes. Oxidative salivary enzymes inhibit or limit the growth of oral pathogens, thereby supporting the natural host defense system; polysaccharide hydrolases (mutanases and dextranases) degrade important carbohydrate components of the biofilm matrix, whereas proteases disrupt bacterial adhesion to oral surfaces or affect cell-cell interactions. The efficiency of the enzymes in in vitro and in vivo studies, advantages and limitations, as well as future perspectives for improving the enzymatic strategy are discussed.

Theranostic Imaging of Cancer Gene Therapy.

Gene-directed enzyme prodrug therapy (GDEPT) is a promising therapeutic approach for treating cancers of various phenotypes. This strategy is independent of various other chemotherapeutic drugs used for treating cancers where the drugs are mainly designed to target endogenous cellular mechanisms, which are different in various cancer subtypes. In GDEPT an external enzyme, which is different from the cellular proteins, is expressed to convert the injected prodrug in to a toxic metabolite, that normally kill cancer cells express this protein. Theranostic imaging is an approach used to directly monitor the expression of these gene therapy enzymes while evaluating therapeutic effect. We recently developed a dual-GDEPT system where we combined mutant human herpes simplex thymidine kinase (HSV1sr39TK) and E. coli nitroreductase (NTR) enzyme, to improve therapeutic efficiency of cancer gene therapy by simultaneously injecting two prodrugs at a lower dose. In this approach we use two different prodrugs such as ganciclovir (GCV) and CB1954 to target two different cellular mechanisms to kill cancer cells. The developed dual GDEPT system was highly efficacious than that of either of the system used independently. In this chapter, we describe the complete protocol involved for in vitro and in vivo imaging of therapeutic cancer gene therapy evaluation.

Substitution therapy enzymes immobilized on nutritive fibers

Disturbances induced by substitution therapy of chronic pancreatic diseases are eliminated with the aid of various en- zyme preparations, primarily digestive hydrolases. Most promising of these are drugs containing a combination of en- zymes capable of hydrolyzing proteins, fats, and carbohy- drates. However, the efficiency of these drugs is frequently reduced by deactivation of the substitution therapy enzymes in the gastrointestinal tract, in particular, under the action of acidic gastric medium and temperature factors [1]. A possible way to remove this disadvantage of the drugs for substitution therapy is provided by enzymes immobilized on an appropriate carrier. In this connection, it would be of interest to consider the possibility to use nutritive fibers (NFs)-a combination of natural polymers with the main components including lig- nins, pectins, hemicellulose, and cellulose [2]. This choice is additionally justified by the fact that the NF proper are known to serve as regulators of metabolic processes and digestive functions in the organism. In this work we have examined the possibility to immobi- lize alkaline protease, 13-galactosidase, oraze, and solizym on a NF cartier. Our purpose was to obtain the immobilized form of not only individual enzymes, but of their combina- tion as well, and to study the functioning of NF-immobilized enzymes. EXPERIMENTAL PART The enzymatic activity was studied in duodenum of rats narcotized with urethane (1 g/kg). After opening the ab- dominal cavity, a part of the intestine was ligated and a sub- strate mixture (2 ml) with a suspension of immobilized en- zymes (0.5 ml) was cannulated. In the reference sample, the enzyme was replaced by an enzyme substance solution. All drug preparations were introduced at a dose of 25 mg/kg.

The promise of enzymes in therapy of hyperlipidemia

Treatment of hyperlipidemias must be commenced in that intestinal segment where alimentary lipoprotein aggregate generation is initiated postprandially and the aggregates are still in nascent form. This is to prevent the formation of potentially pathological units that do not equilibrate with the blood colloid. The colloid chemical state of the alimentary lipoprotein entities in the systemic circulation is decisive for maintenance of thair stability and for their disposal from the blood. The causes of impaired, “unripe,” lipoprotein aggregate formation include: (a) the action of human pancreatic lipase is even normally a restricted and vulnerable process, (b) inadequate ratio of protein to fat to stabilize the aggregates, (c) defective proteolysis in the small intestine, and (d) the dual behavior of the surface tension-lowering agent, the lecithin antagonist cholesterol. On the one hand, cholesterol represents physicochemically the weakest link in the lipoprotein interfaces. On the other hand, an excess of cholesterol in lipoprotein interfaces decreases the stability of the surface film. This leads to changes in the decay of the entities and the cell surface-active lipid constituents of the “unripe” plasma lipoprotein colloid complexes can easily be adsorbed to the endothelial cell surface plasma membrane: the excess aggregate-cholesterol molecules simply join, or dissolve in, the cell membrane-cholesterol which thus acts as its own receptor or solvent: cholesterol dimers occur. This causes rigidity of the endothelial cell surface membrane and leads to impaired cell metabolism. The intima recognizes the impairment as a foreign body and initiates a physiological defense reaction including phagocytosis. The event may be still more dangerous if the adrenergic receptors of the autonomous nervous system involved in lipolysis with subsequent plasma efflux are simultaneously stimulated. The therapeutic measure indicated is production of balanced alimentary lipoprotein aggregates. This can be achieved by oral administration of (a) protease, and (b) non-specific micellar lipase. The lipase functions without restriction and can perform all the activities that human pancreatic lipase cannot. The enzymes utilized are of mold origin and generally available.

The promise of enzymes in therapy of uremia. III. Enzyme preparation.

The promise of enzymes in therapy of uremia. III. Enzyme preparation.

The promise of enzymes in therapy of uremia. II. Theoretical basis--enzyme properties.

Apres un rappel sur certaines proprietes generales des enzymes on discute les micro-organismes avec des systemes enzymatiques adaptatifs (entrainables), les proteines des dechets, l'azote non proteique comme seule source d'azote pour les vaches laitieres, la capacite des enzymes microbiennes a decomposer l'azote non proteique