RESUMEN
Chemical catalysts are being replaced by biocatalysts in almost all industrial applications due to environmental concerns, thereby increasing their demand. Enzymes used in current industries are produced in microbial systems or plant seeds. We report here five newly launched leaf-enzyme products and their validation with 15 commercial microbial-enzyme products, for detergent or textile industries. Enzymes expressed in chloroplasts are functional at broad pH/temperature ranges as crude-leaf extracts, while most purified commercial enzymes showed significant loss at alkaline pH or higher temperature, required for broad range commercial applications. In contrast to commercial liquid enzymes requiring cold storage/transportation, chloroplast enzymes as a leaf powder can be stored up to 16 months at ambient temperature without loss of enzyme activity. Chloroplast-derived enzymes are stable in crude-leaf extracts without addition of protease inhibitors. Leaf lipase/mannanase crude extracts removed chocolate or mustard oil stains effectively at both low and high temperatures. Moreover, leaf lipase or mannanase crude-extracts removed stain more efficiently at 70 °C than commercial microbial enzymes (<10% activity). Endoglucanase and exoglucanase in crude leaf extracts removed dye efficiently from denim surface and depilled knitted fabric by removal of horizontal fibre strands. Due to an increased demand for enzymes in the food industry, marker-free lettuce plants expressing lipase or cellobiohydrolase were created for the first time and site-specific transgene integration/homoplasmy was confirmed by Southern blots. Thus, leaf-production platform offers a novel low-cost approach by the elimination of fermentation, purification, concentration, formulation and cold-chain storage/transportation. This is the first report of commercially launched protein products made in leaves and validated with current commercial products.
Asunto(s)
Detergentes , Lipasa , Hojas de la Planta , Industria Textil , Cloroplastos/enzimología , Cloroplastos/genética , Detergentes/normas , Estabilidad de Enzimas , Lipasa/genética , Lipasa/aislamiento & purificación , Lipasa/metabolismo , Lipasa/normas , Hojas de la Planta/enzimología , Hojas de la Planta/genética , Temperatura , Industria Textil/métodos , beta-Manosidasa/genética , beta-Manosidasa/aislamiento & purificación , beta-Manosidasa/metabolismo , beta-Manosidasa/normasRESUMEN
PURPOSE: Pancreatic enzyme replacement therapy (PERT) involves exogenous enzyme supplementation and is used in the treatment of pancreatic exocrine insufficiency. Clinical efficacy of PERT preparations is a function of physical properties and release kinetics that vary between commercially available products. In this study, we evaluated the physical properties, in vitro dissolution, and release kinetics of commercially available pancreatic enzyme preparations available in the Indian market. METHODS: Physical properties such as particle size distribution and water content of the capsules were measured by dynamic light scattering and Karl-Fischer titration method, respectively. An analytical procedure based on the European pharmacopoeia (EP) method was used to determine lipase activity, and a modified United States pharmacopoeia (USP)-based method was used for dissolution studies. Enzyme release was ascertained under gastroduodenal conditions in buffered media. RESULTS: Considerable variations in physical properties such as particle size and water content were observed between pancreatic enzyme preparations. Some preparations failed to meet the labeled lipase content as per USP standards (>90% label claim) and showed inconsistent release behavior (>5% relative standard deviation). CONCLUSION: Differences exist between pancreatic enzyme preparations in terms of physical properties, dissolution, and release behavior that can affect their clinical efficacy. The present study suggests, therefore, that these preparations should not be used interchangeably.
Asunto(s)
Fármacos Gastrointestinales/análisis , Lipasa/análisis , Pancreatina/análisis , Cápsulas , Liberación de Fármacos , Terapia de Reemplazo Enzimático , Fármacos Gastrointestinales/química , Fármacos Gastrointestinales/normas , Humanos , India , Lipasa/química , Lipasa/normas , Pancreatina/química , Pancreatina/normas , Tamaño de la PartículaRESUMEN
CONTEXT: Acute pancreatitis is diagnosed on the basis of clinical features, biochemical tests and imaging studies. Normal serum amylase level has been reported in the setting of acute pancreatitis but normal serum lipase level in acute pancreatitis is extremely rare. CASE REPORT: Herein, we present a case series of acute pancreatitis with normal serum lipase levels along with a review of the topic. CONCLUSION: In appropriate clinical setting, the diagnosis of acute pancreatitis should be entertained even with normal serum amylase and lipase levels.
Asunto(s)
Lipasa/sangre , Pancreatitis/sangre , Pancreatitis/diagnóstico , Enfermedad Aguda , Adulto , Anciano , Femenino , Humanos , Lipasa/normas , Masculino , Persona de Mediana Edad , Valores de ReferenciaRESUMEN
Objectives of this work are to study changes in the immunoreactive HTGL mass during storage under various conditions. In addition, the shelf-life of the HTGL ELISA kit was confirmed. The immunological reactivity of HTGL in PHP stored in the liquid, frozen, or lyophilized state was monitored using purified human PHP-HTGL as the standard material. Furthermore, the long-term stability of the HTGL ELISA kit was ascertained. The immunoreactive HTGL mass in the lyophilized PHP maintained its initial immunological reactivity for at least 26 months at 4 degrees C or lower. The other reagents included in the HTGL ELISA kit also have a long shelf-life when they are stored at 4 degrees C or less. HTGL in PHP was stabilized by lyophilization and can be used as the standard material for HTGL ELISA; the HTGL ELISA kit has a long shelf-life, i.e., more than two years.
Asunto(s)
Ensayo de Inmunoadsorción Enzimática/métodos , Lipasa/análisis , Anticuerpos Monoclonales/inmunología , Estabilidad de Medicamentos , Almacenaje de Medicamentos , Liofilización , Humanos , Lipasa/inmunología , Lipasa/normas , Hígado/enzimología , Juego de Reactivos para Diagnóstico , Temperatura , Factores de TiempoRESUMEN
BACKGROUND: To effectively assess and correct for intermethod variability, calibration and control materials (CCMs) must show the same intermethod behavior as patient sera, i.e., they must be commutable. We describe the commutability of selected CCMs for lipase assays, the impact of noncommutability of CCMs in normalizing patient results, and characteristics of reagents that affect assay specificity and commutability. METHODS: Lipase was measured in 98 patient sera and in 29 commercial CCMs, with 2 commercial methods using different substrates and with 4 experimental methods using 1,2-o-dilauryl-rac-glycero-3-glutaric acid-(6'-methylresorufin) ester as substrate and colipase as cofactor, but differing in the stabilizing proteins used and in the size of the substrate micelles. RESULTS: The noncommutability rate, i.e., the frequency of aberrant intermethod behavior of CCMs in comparison with patient sera, was 27% for liquid CCMs and 47% for lyophilized CCMs. The normalized residuals, measuring the degree of noncommutability, were -2.3 to 2.4 for CCMs with "normal" lipase activity, and -3.5 to 21.7 for CCMs with higher lipase activity. Recalculation of patient results with CCMs as calibrators decreased or increased the original bias according to whether the CCMs were commutable. CONCLUSIONS: For the lipase methods in this study, the frequency of noncommutability of CCMs is affected by assay-specific characteristics, including size of substrate micelles and the presence or absence of added proteins.
Asunto(s)
Lipasa/normas , Calibración , Humanos , Indicadores y Reactivos , Modelos Lineales , Lipasa/sangre , Estándares de Referencia , Sensibilidad y EspecificidadRESUMEN
We have carried out an independent investigation of the lipase activity of several pancreatin preparations, including high-lipase preparations of pancreatin that have recently become available. Six preparations from three pharmaceutical companies, Cilag, Duphar, and Merck, were analyzed during the recommended shelf life of the preparation and included a standard and a high-lipase preparation from each manufacturer. All preparations studied showed lipase activity in excess of that which was stated on the packets, and in some cases it was more than twofold. This has important implications for patients and prescribers since changes in apparent enzyme requirements may reflect differences in the potency of batches with time. As it is impossible to evaluate capsule requirements with each new batch prescribed, the presence or relief of symptoms of malabsorption will probably continue to be the way in which most patients monitor their enzyme supplementation on a regular basis. We believe that clinicians need to be aware of the extent of possible intrinsic variation between individual prescriptions before attempting to define possible dose/symptom relationships.
Asunto(s)
Alimentos Fortificados/análisis , Lipasa/análisis , Páncreas/enzimología , Pancreatina/química , Relación Dosis-Respuesta a Droga , Industria Farmacéutica/normas , Insuficiencia Pancreática Exocrina/tratamiento farmacológico , Humanos , Lipasa/normas , Lipasa/uso terapéutico , Síndromes de Malabsorción/tratamiento farmacológico , Pancreatina/uso terapéutico , Control de Calidad , Estándares de ReferenciaRESUMEN
In-vitro activity of 14 commercial pancreatin preparations, commonly used in the Federal Republic of Germany, were tested. All had been declared by their manufacturers to contain more than 6000 FIP (Fédération International Pharmaceutique) units of lipase and to be acid resistant. The declared lipase and amylase amounts were found to be present in 11 of the 14 preparations. Three of the 14 preparations, said to be acid resistant were found not to be so in buffer with falling pH values between 4.0 and 2.5, so that there occurred an, at times marked, loss of enzyme activity. Most noticeable was the poor solubility of most preparations at pH 6.6. Only three of the 14 liberated their total enzyme content within 60 minutes, as they should for theoretical reasons, based on the relatively short duodeno-cecal transit time.
Asunto(s)
Extractos Pancreáticos/normas , Amilasas/análisis , Amilasas/normas , Bromelaínas/análisis , Bromelaínas/normas , Ácido Deshidrocólico/análisis , Ácido Deshidrocólico/normas , Dimetilpolisiloxanos/análisis , Dimetilpolisiloxanos/normas , Combinación de Medicamentos/análisis , Combinación de Medicamentos/normas , Concentración de Iones de Hidrógeno , Lipasa/análisis , Lipasa/normas , Extractos Pancreáticos/análisis , Pancreatina/análisis , Pancreatina/normas , Solubilidad , Tripsina/análisis , Tripsina/normasRESUMEN
The so called "enzymatic preparations" Rheumajecta and Vasolastine (R & V) belong to the complementary treatments. The preparations have been used for many years in the treatment of patients with rheumatic conditions such as rheumatoid arthritis (RA) in the Netherlands and other countries of Europe, although a proper study showing efficacy was never performed. Therefore a double-blind, placebo-controlled, modified cross-over trial during two periods of 3 months was performed in 34 patients with RA. They were allocated at random to R & V or to placebo injections, all intramuscular. After 3 months of therapy each patient could opt for cross-over in the event of lack of subjective improvement. Clinical assessments including Ritchie's articular index, grip strength, the DUTCH-AIMS questionnaire, ESR and CRP were performed. R & V did not prove to be more effective than placebo. No serious side-effects were seen.
Asunto(s)
Artritis Reumatoide/tratamiento farmacológico , Catalasa/uso terapéutico , Colina O-Acetiltransferasa/uso terapéutico , Lipasa/uso terapéutico , Lipooxigenasa/uso terapéutico , Sulfato Adenililtransferasa/uso terapéutico , Sulfurtransferasas/uso terapéutico , Adolescente , Adulto , Anciano , Artritis Reumatoide/patología , Artritis Reumatoide/fisiopatología , Catalasa/administración & dosificación , Catalasa/normas , Colina O-Acetiltransferasa/administración & dosificación , Colina O-Acetiltransferasa/normas , Método Doble Ciego , Combinación de Medicamentos , Quimioterapia Combinada , Femenino , Humanos , Inyecciones Intramusculares , Lipasa/administración & dosificación , Lipasa/normas , Lipooxigenasa/administración & dosificación , Lipooxigenasa/normas , Masculino , Persona de Mediana Edad , Índice de Severidad de la Enfermedad , Sulfato Adenililtransferasa/administración & dosificación , Sulfato Adenililtransferasa/normas , Sulfurtransferasas/administración & dosificación , Sulfurtransferasas/normas , Encuestas y CuestionariosRESUMEN
Treatment of steatorrhea by lipase supplementation therapy has become more successful in the last decade due to better understanding of the physiology and pathophysiology of the digestive process. Porcine lipase has been the therapeutic standard for several decades and will continue to be the treatment of choice in pancreatic exocrine insufficiency. Modern therapeutic concepts recommend administration of 25,000-40,000 units of porcine lipase per meal using pH-sensitive pancreatin microspheres. In case of treatment failure, the dose should be increased, compliance should be checked, and other reasons for malabsorption should be excluded. Still, in most patients, lipid digestion cannot be completely normalized by current standard therapy, and future developments are needed for optimizing treatment. In this article, pathophysiologic characteristics of pancreatic exocrine insufficiency, prerequisites for use of alternative lipase sources as well as currently available lipases of nonporcine origin, and new developments are discussed. Current literature suggests that bovine lipase products present a theoretical alternative but play no major role in the western world. Fungal lipase has inferior properties compared with conventional products. Bacterial lipase products show promising potential and offer future therapeutic alternatives. Moreover, human pancreatic lipase gene transfer and application of bioengineered human gastric lipase appear on the horizon.