RESUMEN
BACKGROUND AND AIMS: Shoulder capsular surgery is nowadays usually performed arthroscopically, and the proportion of arthroscopic method has rapidly increased during the last two decades. We assessed the incidence of shoulder capsular surgery procedures in Finland between 1999 and 2008. MATERIAL AND METHODS: We gathered the shoulder capsular surgery procedures for all kinds of shoulder instability in Finland between 1999 and 2008 from National Hospital Discharge Register and limited the patient material to include only certain diagnosis (International Classification of Diseases, 10th Edition) and Nordic Medico-Statistical Committee procedure code combinations. We analyzed the data in the whole country, between different age groups, and in university hospital districts. RESULTS: The total incidence of shoulder capsular surgery procedures in Finland increased from 17 to 33 per 100,000 person-years. The incidence of arthroscopic procedures increased from 11 to 30 per 100,000 person-years and the proportion of arthroscopic procedures increased from 63% to 92% between years 1999 and 2007. The incidence of shoulder capsular surgery procedures increased on average around 90% in almost all age groups and particularly in the older age groups. We observed no significant geographical variation between university hospital districts. CONCLUSION: The incidence of shoulder capsular surgery procedures increased on average round 90% in almost all age groups. It seems to be difficult to support the rapidly increased rates of shoulder capsular surgery procedures or the arthroscopic method based on scientific evidence. While also older patients are treated with shoulder capsular surgery, well-defined indications for surgical intervention are needed so that the operations are conducted for the symptomatic patients benefitting most regardless of patients' age.
Asunto(s)
Artroscopía/estadística & datos numéricos , Cápsula Articular/cirugía , Inestabilidad de la Articulación/cirugía , Sistema de Registros , Articulación del Hombro/cirugía , Adolescente , Adulto , Niño , Femenino , Finlandia/epidemiología , Humanos , Incidencia , Inestabilidad de la Articulación/diagnóstico , Inestabilidad de la Articulación/epidemiología , Masculino , Persona de Mediana Edad , Estudios Retrospectivos , Adulto JovenRESUMEN
BACKGROUND AND AIMS: The indications for operative treatment of lateral tibial plateau fractures are still controversial. The objective of this study was to determine whether residual articular surface depression and valgus malalignment of plated lateral tibial plateau fractures at medium-term follow-up affect the clinical and radiographic outcomes. MATERIAL AND METHODS: A chart review of patients with operatively treated (AO type B3.1) tibial plateau fractures that were admitted to our level I trauma center between 2002 and 2008 was performed. Out of 123 patients, 73 were available to participate in a clinical and radiographic follow-up examination. The mean follow-up time was 54 months. Patients were clinically assessed and completed the Lysholm knee score and Western Ontario and McMaster Universities Osteoarthritis Index. Maximal articular surface depression, radiological mechanical axis, and degree of posttraumatic osteoarthritis were evaluated from standing radiographs. RESULTS: Patients with valgus malalignment of 5° or greater at follow-up developed more advanced osteoarthritis (Kellgren-Lawrence grade 3-4) than patients with a normal mechanical axis (p = 0.006). Similarly, patients with articular depression greater than 2 mm at follow-up also developed more advanced osteoarthritis compared to patients with a depression of 2 mm or less (p = 0.001). The degree of valgus malalignment or articular depression had no effect on the Western Ontario and McMaster Universities Osteoarthritis Index or Lysholm scores. CONCLUSIONS: The postoperative articular congruity and normal mechanical axis of the lower leg after plate fixation in lateral tibial plateau fractures seem to have a role in prevention of posttraumatic osteoarthritis but does not appear to predict clinical outcome at medium-term follow-up. The role of initial dislocation and associated cartilage damage in the development of osteoarthritis following these fractures is still unknown.
Asunto(s)
Fijación Interna de Fracturas , Articulación de la Rodilla/diagnóstico por imagen , Osteoartritis de la Rodilla/etiología , Fracturas de la Tibia/complicaciones , Adolescente , Adulto , Anciano , Femenino , Finlandia/epidemiología , Estudios de Seguimiento , Humanos , Articulación de la Rodilla/fisiopatología , Masculino , Persona de Mediana Edad , Osteoartritis de la Rodilla/diagnóstico por imagen , Osteoartritis de la Rodilla/epidemiología , Pronóstico , Radiografía , Rango del Movimiento Articular , Estudios Retrospectivos , Factores de Riesgo , Fracturas de la Tibia/diagnóstico por imagen , Fracturas de la Tibia/cirugía , Factores de Tiempo , Adulto JovenRESUMEN
The main purpose of this study was to explore how the number of home, sports, and other leisure time injuries among young people aged 15 - 25 years has developed in Finland in 1988 - 2003. In 1988, 2559 people aged 15 - 25 years were interviewed by Statistics Finland. In 1993, 1997 and 2003 those amounts were 751, 1638 and 1382. The subjects were asked in a telephone interview to report the injuries in which they had been involved during the previous 12 months. The crude injury incidence in 2003 was 364 per 1000 person years in men and 246 in women. The total number of injuries increased 49 % between the years 1988 and 2003. The number of sports injuries increased the most. The increase was greater among men than women, and the injury rate was higher among both men and women aged 15 - 19 years than those aged 20 - 25 years. More research is needed on whether and how exercise and time at home have changed and become riskier among young people. It is also essential to find out which prevention methods are the most effective among young people and implement these measures in a well-planned and targeted manner.
Asunto(s)
Traumatismos en Atletas/epidemiología , Adolescente , Adulto , Factores de Edad , Traumatismos en Atletas/prevención & control , Recolección de Datos , Femenino , Finlandia/epidemiología , Humanos , Masculino , Factores de Riesgo , Factores Sexuales , Encuestas y CuestionariosRESUMEN
Tyramine, histamine, putrescine and cadaverine, the most common biogenic amines indicating the food quality, were studied in the transglutaminase-catalyzed reaction. Transglutaminase (protein-glutamine gamma-glutamyltransferase EC 2.3.2.13) catalyzes an acyl transfer reaction between a donor substrate and an acceptor substrate (e.g. biogenic amine) and forms a cross-linkage between substrates with a release of ammonia. The reaction can be monitored by measuring the ammonia produced in the reaction. The concentration of produced ammonia was found to be proportional to the concentration of biogenic amine and could hence be used to determination of biogenic amines in food matrixes.
RESUMEN
In this paper the oxidation of milled wood lignin (MWL), catalysed by three enzymes, i.e. laccase, tyrosinase and horseradish peroxidase (HRP) was studied. The oxidation was followed by measuring the consumption of O(2) during laccase and tyrosinase treatment and of H(2)O(2) during HRP treatment. Both laccase and HRP were found to oxidise lignin effectively, whereas the effect of tyrosinase was negligible. The changes in MWL molecular-weight distributions caused in the reactions were analysed by gel permeation chromatography. Both laccase and HRP treatments were found to polymerise MWL. Peroxidase treatment was found to decrease the amount of phenolic hydroxyls in MWL, whereas no such effect could be detected in the laccase-treated sample. Both laccase and HRP treatments were, however, found to increase the amount of conjugated structures in MWL. The formation of phenoxy radicals during the treatments was studied by electron paramagnetic resonance spectroscopy. Phenoxy radicals were detected in both laccase and HRP-treated samples. The amount of the formed phenoxy radicals was found to be essentially constant during the detected time (i.e. 20-120 min after the addition of enzyme).
Asunto(s)
Peroxidasa de Rábano Silvestre/metabolismo , Lacasa/metabolismo , Lignina/metabolismo , Monofenol Monooxigenasa/metabolismo , Madera , Biotecnología/métodos , Peróxido de Hidrógeno/metabolismo , Oxidación-Reducción , Oxígeno/metabolismoRESUMEN
Overuse tendon injury is one of the most common injuries in sports. The etiology as well as the pathophysiological mechanisms leading to tendinopathy are of crucial medical importance. At the moment intrinsic and extrinsic factors are assumed as mechanisms of overuse tendon injury. Except for the acute, extrinsic trauma, the chronic overuse tendon injury is a multifactorial process. There are many other factors, such as local hypoxia, less of nutrition, impaired metabolism and local inflammatory that may also contribute to the development of tissue damage. The exact interaction of these factors cannot be explained entirely at the moment. Further studies will be necessary in order to get more information.
Asunto(s)
Traumatismos en Atletas/fisiopatología , Trastornos de Traumas Acumulados/fisiopatología , Traumatismos de los Tendones/fisiopatología , Tendón Calcáneo/lesiones , Tendón Calcáneo/patología , Tendón Calcáneo/fisiopatología , Traumatismos en Atletas/diagnóstico , Traumatismos en Atletas/etiología , Trastornos de Traumas Acumulados/diagnóstico , Trastornos de Traumas Acumulados/etiología , Humanos , Rotura , Tendinopatía/diagnóstico , Tendinopatía/etiología , Tendinopatía/fisiopatología , Traumatismos de los Tendones/diagnóstico , Tendones/patología , Tendones/fisiopatologíaRESUMEN
Most smokers begin smoking in adolescence. It is less well known how young people quit smoking and the factors that are associated with this process. A 15-year follow-up study on the North Karelia Youth Project has made it possible to assess these factors using a longitudinal study design. The project began in 1978 with students in Grade 7 of junior high school (age 13 years) and concluded in 1980 when the students reached Grade 9 (age 15 years). The follow-up study included four additional surveys. The present analyses are based on the data collected at ages 15, 21 and 28. The original sample comprised 903 students and the response rate of the last survey was 71%. A quarter (26%) of daily smokers and about half (46%) of occasional smokers at age of 15 had quit by the age of 28. The cessation rate was higher among females than males (P = 0.006). The cessation rate was higher among married (P = 0.015), employed (P = 0.01) and white-collar workers (P = 0.006). Cessation was less prevalent among those who had friends (P < 0.001) and family (P = 0.012) members who smoked. The cessation rate was lower among those who consumed fatty milk (P = 0.050), had less leisure-time physical activity (P = 0.032) and consumed more alcohol (P < 0.001). One-third of all teenage smokers stop smoking before the age of 28, averaging a 2.3% annual decline. Cessation is greater among occasional than daily smokers and greater overall among females.
Asunto(s)
Cese del Hábito de Fumar/estadística & datos numéricos , Adolescente , Conducta del Adolescente , Adulto , Factores de Edad , Femenino , Finlandia/epidemiología , Humanos , Estudios Longitudinales , Masculino , Factores de Riesgo , Encuestas y CuestionariosRESUMEN
The Achilles tendon is the strongest tendon in the human body. Because most Achilles tendon injuries take place in sports and there has been an common upsurge in sporting activities, the number and incidence of the Achilles tendon overuse injuries and complete ruptures have increased in the industrialized countries during the last decades. The most common clinical diagnosis of Achilles overuse injuries is tendinopathy, which is characterized by a combination of pain and swelling in the Achilles tendon accompanied by impaired ability to perform strenuous activities. Most patients with Achilles tendon injury respond favorably to conservative treatment and only those who fail to respond to carefully followed nonoperative treatment should undergo surgery for repair. A complete rupture of the Achilles tendon usually occurs in sports that require jumping, running, and quick turns. Although histopathologic studies have shown that ruptured Achilles tendons include clear degenerative changes before the rupture, many of the Achilles tendon ruptures occur suddenly without any preceding signs or symptoms. Neither conservative nor operative treatment is a treatment of choice for the ruptured Achilles tendon. It is generally accepted that surgery should be performed on ruptured Achilles tendons in young, physically active patients and in those patients for whom the diagnosis or the treatment of the rupture has been delayed, whereas the results of conservative treatment are an acceptable outcome in older patients with sedentary lifestyles. Many important issues still remain unanswered concerning the cause, pathogenesis, diagnosis, and management of the Achilles tendon disorders. Only when these issues have been solved by well-controlled studies can tailored treatment protocols be created.
Asunto(s)
Tendón Calcáneo/lesiones , Tendón Calcáneo/fisiopatología , Tendón Calcáneo/cirugía , Artritis Reumatoide/complicaciones , Traumatismos en Atletas/etiología , Traumatismos en Atletas/fisiopatología , Traumatismos en Atletas/terapia , Humanos , Dolor/fisiopatología , Rotura , Rotura EspontáneaRESUMEN
To determine the long-term outcome of patients treated nonoperatively for acute or subchronic (duration of the symptoms before initiation of the treatment less than 6 months) Achilles tendinopathy, we performed a follow-up analysis on 83 of 107 patients an average 8 +/- 2 (SD) years after the initial contact. The analysis included a questionnaire, clinical examination, performance tests, muscle strength measurement, and ultrasonographic examination. Twenty-four of the 83 patients (29%) had to be operated on during the follow-up period. Seventy patients (84%) had full recovery of their activity level, and at 8 years' follow-up 78 patients (94%) were asymptomatic or had only mild pain with strenuous exercise. However, a clear side-to-side difference between the involved and the uninvolved sides was observed on the performance test, clinical examination, and ultrasonography. Also, 34 patients (41%) started to suffer from overuse symptoms in the initially uninvolved Achilles tendon. The results of our 8-year follow-up showed that the long-term prognosis of patients with acute-to-subchronic Achilles tendinopathy is favorable as determined by subjective and functional assessments. In the clinical and ultrasonographic examinations, mild-to-moderate changes were observed rather frequently in both the involved and initially uninvolved Achilles tendons, but the occurrence of these changes was not clearly related to the patients' symptoms.
Asunto(s)
Tendón Calcáneo/lesiones , Traumatismos de los Tendones/patología , Tendón Calcáneo/diagnóstico por imagen , Tendón Calcáneo/patología , Tendón Calcáneo/cirugía , Actividades Cotidianas , Adulto , Ejercicio Físico , Femenino , Estudios de Seguimiento , Humanos , Masculino , Análisis y Desempeño de Tareas , Traumatismos de los Tendones/cirugía , Resultado del Tratamiento , UltrasonografíaRESUMEN
We analyzed the complications after surgical treatment of Achilles tendon overuse injuries in 432 consecutive patients. The patients underwent a clinical examination 2 weeks, and 1, 2, and 5 months after the surgery. If a complication appeared, the patient was followed up clinically for at least 1 year. There were 46 (11%) complications in the 432 patients: 14 skin edge necroses, 11 superficial wound infections, 5 seroma formations, 5 hematomas, 5 fibrotic reactions or scar formations, 4 sural nerve irritations, 1 new partial rupture, and 1 deep vein thrombosis. Fourteen patients with a complication had reoperations: four patients for skin edge necrosis, two for superficial wound infection, two for seroma formation, one for hematoma formation, two for fibrotic reaction or scar formation, two for sural nerve irritation, and one for a new partial rupture. About every 10th patient treated surgically for chronic Achilles tendon overuse injury suffered from a postoperative complication that clearly delayed recovery. However, the majority of patients with a complication healed and returned to their preinjury levels of activity. To reduce this morbidity, it is essential that the surgeon be continuously aware of the possibility of postoperative complications and use proper surgical techniques.
Asunto(s)
Tendón Calcáneo/lesiones , Tendón Calcáneo/cirugía , Traumatismos en Atletas/cirugía , Complicaciones Posoperatorias , Adolescente , Adulto , Anciano , Femenino , Estudios de Seguimiento , Humanos , Masculino , Persona de Mediana Edad , Morbilidad , Ortopedia/métodos , Rotura , Resultado del TratamientoRESUMEN
New types of antimicrobial compounds were identified in the culture filtrate of Lactobacillus plantarum VTT E-78076. Activity was detected in the low molecular mass fraction separated by gel chromatography. This fraction totally inhibited the growth of the Gram-negative test organism, Pantoea agglomerans (Enterobacter agglomerans) VTT E-90396. Characteristic compounds from this fraction were identified by GC/MS-analysis and the identification was confirmed using pure commercial reference compounds in identical chromatographs and in antimicrobial tests. The active fraction included benzoic acid (CAS 65-85-0), 5-methyl-2,4-imidazolidinedione (CAS 616-03-5, methylhydantoin), tetrahydro-4-hydroxy-4-methyl-2H- pyran-2-one (CAS 674-26-0, mevalonolactone) and 3-(2-methylpropyl)-2,5-piperazinedione (CAS 5845-67-0, cyclo(glycyl-L-leucyl)). These compounds in concentrations of 10 ppm inhibited growth of the test organism by 10-15% when acting separately, but 100% when all were applied together with 1% lactic acid. The inhibition was 40% by 1% lactic acid alone. The compounds were also active against Fusarium avenaceum (Gibberella avenacea) VTT-D-80147. The inhibition was 10-15% by separate compounds in concentrations of 10 ppm and maximally 20% in combinations. Fungal growth was not inhibited by lactic acid. Inhibition by unfractionated Lact. plantarum culture filtrate was 37% and by the low molecular mass fraction, 27%.
Asunto(s)
Antiinfecciosos/metabolismo , Enterobacteriaceae/efectos de los fármacos , Fusarium/efectos de los fármacos , Lactobacillus/metabolismo , Antibacterianos , Antiinfecciosos/aislamiento & purificación , Antiinfecciosos/farmacología , Ácido Benzoico/metabolismo , Ácido Benzoico/farmacología , Enterobacteriaceae/crecimiento & desarrollo , Fusarium/crecimiento & desarrollo , Calor , Hidantoínas/metabolismo , Hidantoínas/farmacología , Ácido Láctico/farmacología , Ácido Mevalónico/análogos & derivados , Ácido Mevalónico/metabolismo , Ácido Mevalónico/farmacología , Pruebas de Sensibilidad Microbiana , Neuropéptidos/metabolismo , Neuropéptidos/farmacología , Péptidos Cíclicos/metabolismo , Péptidos Cíclicos/farmacologíaRESUMEN
PURPOSE: To assess the value of US in the diagnosis of various Achilles tendon disorders. MATERIAL AND METHODS: Pre-operative US was compared with surgical findings in 79 patients with an Achilles tendon complaint. RESULTS: US was highly reliable for verifying a complete Achilles tendon rupture, only one false-negative US examination was found in the 26 surgically verified cases. For diagnosing retrocalcanear bursitis, US was accurate: 6 out of the 8 cases of bursitis were found and there were no false-positive cases. There were also no false-positive US findings in patients with peritendinitis/tendinitis, but 7 false-negative US cases among the 40 surgically verified peritendinitis/tendinitis patients indicated that a negative US finding in a clinically suspected case of peritendinitis/tendinitis is unreliable. US also seemed to be inadequate for differentiating partial tendon rupture from a focal tendon degeneration. Nevertheless, the occurrence and location of such a lesion could be adequately determined by US. CONCLUSION: US can reliably be used for locating the Achilles tendon abnormality, estimating its severity, and determining most of the conditions requiring surgical intervention. However, US is not completely reliable for diagnosing peritendinitis and tendinitis, and it cannot be used to differentiate partial tendon ruptures from focal degenerative lesions.
Asunto(s)
Tendón Calcáneo/lesiones , Bursitis/diagnóstico por imagen , Tendinopatía/diagnóstico por imagen , Traumatismos de los Tendones/diagnóstico por imagen , Tendón Calcáneo/diagnóstico por imagen , Tendón Calcáneo/cirugía , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Bursitis/cirugía , Niño , Diagnóstico Diferencial , Errores Diagnósticos , Femenino , Estudios de Seguimiento , Humanos , Masculino , Persona de Mediana Edad , Reproducibilidad de los Resultados , Estudios Retrospectivos , Rotura , Tendinopatía/cirugía , Traumatismos de los Tendones/cirugía , UltrasonografíaRESUMEN
QT dispersion is a measure of heterogeneity in ventricular repolarization. Increased ECG QT dispersion is associated with life-threatening ventricular arrhythmias. We studied if magnetocardiographic (MCG) measures of QT dispersion can separate postmyocardial infarction patients with and without susceptibility to sustained VT. Manual dispersion measurements were compared to a newly adapted automatic QT interval analysis method. Ten patients with a history of sustained VT (VT group) and eight patients without ventricular arrhythmias (Controls) were studied after a remote myocardial infarction. Single-channel MCGs were recorded from 42 locations over the frontal chest area and the signals were averaged. QT dispersion was defined as maximum-minimum or standard deviation of measured QT intervals. VT group showed significantly more QT and JT dispersion than Controls. QTapex dispersions were 127 +/- 26 versus 83 +/- 21 ms (P = 0.004) and QTend dispersions 130 +/- 37 versus 82 +/- 37 ms (P = 0.013), respectively. Automatic method gave comparable values. Their relative differences were 9% for QTapex and 27% for QTend dispersion on average. In conclusion, increased MCG QT interval dispersion seems to be associated with a susceptibility to VT in postmyocardial infarction patients. MCG mapping with automated QT interval analysis may provide a user independent method to detect nonhomogeneity in ventricular repolarization.
Asunto(s)
Electrocardiografía/métodos , Magnetismo , Infarto del Miocardio/complicaciones , Procesamiento de Señales Asistido por Computador , Taquicardia Ventricular/diagnóstico , Algoritmos , Estudios de Casos y Controles , Femenino , Humanos , Masculino , Persona de Mediana Edad , Proyectos Piloto , Reproducibilidad de los Resultados , Taquicardia Ventricular/etiologíaRESUMEN
OBJECTIVES: This study evaluated the long-term effects of a school- and community-based smoking prevention program in Finland. METHODS: Four intervention schools from North Karelia and two control schools from another province were chosen for the evaluation. Students who received the intervention were taught to resist social pressures to smoke. The program began in 1978 with seventh-grade students and ran through 1980, with a 15-year follow-up. In North Karelia, a community-based smoking cessation program for adults was also carried out. RESULTS: Mean lifetime cigarette consumption was 22% lower among program subjects than among control subjects. Smoking and prevalence were lower up to the age of 21. CONCLUSIONS: Long-term smoking prevention effects can be achieved if a school-based program using a social influence model is combined with community and mass media interventions.
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Educación en Salud , Prevención del Hábito de Fumar , Adolescente , Niño , Femenino , Finlandia , Estudios de Seguimiento , Educación en Salud/métodos , Humanos , Masculino , Prevalencia , Servicios de Salud Escolar , Fumar/epidemiología , Encuestas y CuestionariosRESUMEN
We have used in situ hybridization and Northern blot analysis with oligonucleotide probe to characterize the site of pituitary adenylate cyclase-activating polypeptide (PACAP) synthesis in the rat testis. We observed strong hybridization signal in one third of the cross-sections of the seminiferous tubules, whereas some tubules were devoid of hybridization signal, thus suggesting that PACAP mRNA is expressed in a stage-specific manner. More detailed analysis showed that PACAP mRNA was present in round spermatids at stages III-VII of the cycle. Northern blot hybridization to RNAs extracted from samples of seminiferous tubules at different stages of the epithelial cycle confirmed that expression of PACAP mRNA is restricted to specific stages of the cycle. The highest amount of PACAP mRNA was detected at stages V to early -VII of the cycle, whereas very low levels of mRNA were present at stages I-II and IX-XIV. The present results demonstrate that PACAP mRNA is expressed in the developing germ cells. This suggests that PACAP may function as a paracrine or autocrine regulatory factor for the Sertoli and germ cells, with a specific function during early spermiogenesis, shortly before the onset of nuclear elongation, at the last period of haploid gene activity.
Asunto(s)
Neuropéptidos/genética , ARN Mensajero/análisis , Túbulos Seminíferos/química , Animales , Northern Blotting , Regulación de la Expresión Génica , Hibridación in Situ , Masculino , Neuropéptidos/metabolismo , Polipéptido Hipofisario Activador de la Adenilato-Ciclasa , ARN Mensajero/genética , Ratas , Ratas Sprague-Dawley , Túbulos Seminíferos/metabolismo , Espermatogénesis/genética , Espermatogénesis/fisiología , Testículo/química , Testículo/metabolismoRESUMEN
Cellulases from Trichoderma reesei form an enzyme group with a common structural organization. Each cellulase enzyme is composed of two functional domains, the core region containing the active site and the cellulose-binding domain (CBD). To facilitate the specific detection of each domain, monoclonal antibodies (mAb) against cellobiohydrolase I (CBHI), cellobiohydrolase II (CBHII) and endoglucanase I (EGI) were produced. Five mAb were obtained against CBHI, ten against CBHII and eight against EGI. The location of the antigenic epitope for each antibody was mapped by allowing the antibodies to react with truncated cellulases, synthesized from deleted cDNA in Saccharomyces cerevisiae. Proteolytic fragments of Trichoderma cellulases, obtained by papain digestion, were used to confirm the results. Specific antibodies were detected against the core and the CBD epitopes for all three cellulases. Using the truncated enzymes, it was possible to locate the epitopes to a reasonably short region within the protein. To obtain a quantitative assay for each enzyme, a specific mAb against each antigen was chosen, based on the affinity to the corresponding antigen on Western-blot staining and on filter blots of the cellulolytic yeasts. The mAb were used to quantitative the corresponding enzymes in T. reesei culture medium. Specific quantitation of each cellulase enzyme has not been possible by biochemical assays or using polyclonal antibodies, due to their cross-reactions. Now, these mAb can be specifically used to recognize and quantitate different domains of these three important cellulolytic enzymes.
Asunto(s)
Anticuerpos Monoclonales , Celulosa/metabolismo , Glicósido Hidrolasas/inmunología , Trichoderma/enzimología , Anticuerpos Monoclonales/inmunología , Sitios de Unión , Western Blotting , Celulosa 1,4-beta-Celobiosidasa , Epítopos/inmunología , Expresión Génica , Glicósido Hidrolasas/genética , Papaína , Fragmentos de Péptidos/inmunología , Saccharomyces cerevisiae/enzimología , Saccharomyces cerevisiae/genéticaRESUMEN
Isoelectric focusing is introduced as a technique for the analysis of macromolecular lignin. The analysis is performed in a pH gradient from 3.5 to 10. Separated lignin fragments are visualized under uv light or by silver staining. The method can be used to distinguish between differently processed lignin preparations and to identify their components. Even the slight modification resulting from attack by ligninolytic enzymes could be detected.
Asunto(s)
Focalización Isoeléctrica/métodos , Lignina/aislamiento & purificación , Electroforesis en Gel de Poliacrilamida/métodos , Peroxidasas , Plata , Espectrofotometría Infrarroja , Coloración y Etiquetado , Rayos UltravioletaRESUMEN
We have isolated and characterized a gene coding for the laccase of the lignin-degrading fungus Phlebia radiata. The gene has nine introns and recognizable fungal promoter elements. Sequences homologous to the consensus eukaryotic heat-shock regulatory element can be found in the promoter. RNA hybridization results indicate that this gene is regulated at the transcriptional level. The derived laccase amino acid sequence shows homology to plant ascorbate oxidases, suggesting that the basic structure of the laccase is similar to the three-fold repeated beta-barrel of the ascorbate oxidases. Potential copper ligands and a residue carrying the prosthetic group pyrroloquinoline quinone (PQQ) in the laccase protein can be identified by homology. The intron/exon structure of the laccase gene suggests that this protein could have evolved by exon shuffling.
Asunto(s)
Basidiomycota/genética , Genes Fúngicos , Oxidorreductasas/genética , Secuencia de Aminoácidos , Ascorbato Oxidasa/genética , Secuencia de Bases , Basidiomycota/enzimología , Evolución Biológica , Northern Blotting , Cobre/metabolismo , Exones/genética , Regulación Fúngica de la Expresión Génica/genética , Intrones/genética , Lacasa , Datos de Secuencia Molecular , Oxidorreductasas/química , Oxidorreductasas/metabolismo , Regiones Promotoras Genéticas/genética , Conformación Proteica , Secuencias Reguladoras de Ácidos Nucleicos/genética , Homología de Secuencia de Ácido NucleicoRESUMEN
Extracellular laccase (benzenediol: oxygen oxidoreductase EC 1.10.3.2) from the lignin-degrading fungus, Phlebia radiata, was shown to contain a novel combination of electron carriers as its prosthetic groups. In addition to two copper atoms per enzyme molecule, one molecule of PQQ was included as a cofactor. The EPR spectrum exhibits features of type 1 and type 2 copper atoms. In the enzymatic reaction 4 molecules of lignin model compound, coniferyl alcohol, are oxidized per molecule of oxygen reduced to water. During the reaction coniferyl alcohol is transformed to dilignols.