The author has studied the beta-glucuronidase activity in several tissues such as liver, stomach and small intestine of the male and female rabbits infected with different doses of metacercariae of Clonorchis sinensis. The metacercariae of Clonorchis sinensis were isolated from Pseudorasbora parva caught in Kim Hae by digestion technic. The experimental animals were sacrificed in the period of 1, 7, 14, 21, 28 and 35th days following the infection. The results obtained were summarized as follows: In the groups infected with 100 metacercariae, Beta-glucuronidase activity was slightly increased during the entire periods than control rabbits. It was the highest in the first day with 1.535 and 1.421 mu/g, 14th days with 2.521 and 2.200 mu/g, and then lowered by the time, gradually. In the groups infected with 500 metacercariae, Beta-glucuronidase activity was highly increased on the first day with 1.535 and 1.866 mu/g than that 100 metacercariae groups according to each organs. It was the highest on the 7th day and 14th day. In the groups infected with 1,000 metacercariae, beta-glucuronidase activity was remarkably increased in the first and 14th days according to each organs, and then lowered gradually day by day. beta-glucuronidase activity of all organs was more increased than that of normal organs and the highest activity in the liver with 2.521 mu/g, intestine(1.612) and stomach (1.581) respectively. beta-glucuronidase activity of rabbits was higher in the female than in the male. On the basis of these results, it was suggested that beta-glucuronidase activity was affected by the duration of infection and by the number of Clonorchis sinensis, according to the organs and sex of the rabbits.
parasitology-helminth-trematoda ; Clonorchis sinensis ; beta-glucuronidase ; biochemistry

OBJECTIVETo explore the incidence of various types of mucopolysaccharidosis (MPS) and their clinical characteristics.

METHODSA total of 75 children highly suspected as having MPS underwent quantitative and electrophoretic analysis of urinary glycosaminoglycans (GAGs) and enzymatic analysis of seven types of MPS from January 2009 to December 2011. Fluorescence assay was used to measure the activities of α-L-iduronidase, iduronate-2-sulfatase, α-N-acetylglucosaminidase, galactosamine-6-sulfatase, β-galactosidase, arylsulfatase B and β-glucuronidase in the white blood cells.

RESULTSA total of 52 cases were confirmed with MPS based on clinical, radiological, and enzymatic examinations. The 52 cases, with a mean age of 4.0 ± 2.2 years, included 5 cases of MPS I (10%), 20 cases of MPS II (38%), 20 cases of MPS IVA (38%), 6 cases of MPS VI (12%) and 1 case of MPS VII (2%). No MPS IV B cases or MPS IIIB cases were found. Compared with healthy children of the same age, the GAG/Cr ratio was significantly elevated in 50 confirmed cases of MPS (two MPS IVA cases having no increased ratio). All children with increased urinary GAGs had a confirmed diagnosis of MPS. The age of onset was between 1 and 2 years after birth in most cases, and often complicated by hernia and valvular heart disease. Children with MPS I, MPS II, and MPS VI presented with ugly and unsmooth face, short stature, joint stiffness, and limitation of motion, while children with MPS IVA presented with short stature, skeletal dysplasia, and joint laxity.

CONCLUSIONSType IVA and type II are the most common in MPS cases, followed by type VI and type I. MPS children are characterized by special appearances including ugly and unsmooth facial appearance, short stature and skeletal dysplasia. Quantitative analysis of urinary GAG, as a simple, rapid, and reliable method, is recommended for screening of MPS.

Acetylglucosaminidase ; blood ; Child ; Child, Preschool ; Creatinine ; urine ; Female ; Glucuronidase ; blood ; Glycosaminoglycans ; urine ; Humans ; Iduronidase ; blood ; Infant ; Magnetic Resonance Imaging ; Male ; Mucopolysaccharidoses ; diagnosis ; enzymology ; pathology ; beta-Galactosidase ; blood

β-Glucosidase activity assays constitute an important indicator for the early diagnosis of neonatal necrotizing enterocolitis and qualitative changes in medicinal plants. The drawbacks of the existing methods are high consumption of both time and reagents, complexity in operation, and requirement of expensive instruments and highly trained personnel. The present study provides a simplified, highly selective, and miniaturized glucometer-based strategy for the detection of β-glucosidase activity. Single-factor experiments showed that optimum β-glucosidase activity was exhibited at 50 °C and pH 5.0 in a citric acid-sodium citrate buffer when reacting with 0.03 g/mL salicin for 30 min. The procedure for detection was simplified without the need of a chromogenic reaction. Validation of the analytical method demonstrated that the accuracy, precision, repeatability, stability, and durability were good. The linear ranges of β-glucosidase in a buffer solution and rat serum were 0.0873-1.5498 U/mL and 0.4076-2.9019 U/mL, respectively. The proposed method was free from interference from β-dextranase, snailase, β-galactosidase, hemicellulase, and glucuronic acid released by baicalin. This demonstrated that the proposed assay had a higher selectivity than the conventional dinitrosalicylic acid (DNS) assay because of the specificity for salicin and unique recognition of glucose by a personal glucose meter. Miniaturization of the method resulted in a microassay for β-glucosidase activity. The easy-to-operate method was successfully used to detect a series of β-glucosidases extracted from bitter almonds and cultured by Aspergillus niger. In addition, the simplified and miniaturized glucometer-based assay has potential application in the point-of-care testing of β-glucosidase in many fields, including medical diagnostics, food safety, and environmental monitoring.
Animals ; Aspergillus niger ; Calibration ; Cellulase/analysis* ; Chemistry, Clinical/methods* ; Dextranase/analysis* ; Enterocolitis, Necrotizing/diagnosis* ; Equipment Design ; Flavonoids/analysis* ; Glucose/analysis* ; Glucuronic Acid/analysis* ; Glucuronidase/analysis* ; Glycoside Hydrolases/analysis* ; Hydrogen-Ion Concentration ; Linear Models ; Multienzyme Complexes/analysis* ; Plants, Medicinal ; Polygalacturonase/analysis* ; Rats ; Reproducibility of Results ; beta-Galactosidase/analysis* ; beta-Glucosidase/analysis*

No abstract available.
Breast* ; Glucuronidase* ; Lipase* ; Milk, Human*

The effectiveness of three kinds of enzymes (chitinase, beta-glucuronidase, and lysing enzyme complex), employed as elicitors to enhance the beta-glucan content in the sawdust-based cultivation of cauliflower mushroom (Sparassis latifolia), was examined. The elicitors were applied to the cauliflower mushroom after primordium formation, by spraying the enzyme solutions at three different levels on the sawdust-based medium. Mycelial growth was fully accomplished by the treatments, but the metabolic process during the growth of fruiting bodies was affected. The application of a lysing enzyme resulted in an increase in the beta-glucan concentration by up to 31% compared to that of the control. However, the treatment resulted in a decrease in mushroom yield, which necessitated the need to evaluate its economic efficiency. Although we still need to develop a more efficient way for using elicitors to enhance functional metabolites in mushroom cultivation, the results indicate that the elicitation technique can be applied in the cultivation of medicinal/edible mushrooms.
Agaricales* ; Brassica* ; Fruit ; Glucuronidase ; Metabolism

The aim of this investigation was to study the effect of orthodontic force on the flow of gingival crevicular fluid and activities of arylsulfatase and brta-glucuronidase in crevicular fluid. The material consisted of 12 persons between the ages of 13 years and 22 years and all were categorized Class I, 4-4 extraction cases Crevicular fluids were sampled from distal crevis of each canine before treatment (phase 1), after bracketing (phase 2), after application of force (phase 3) and after run out of orthodontic force (phase 4). Crevicular fluid flow did not show any significant changes during the period of treatment. The activities of arylsulfatase increased significantly after setting of orthodontic appliance without application of force, but did not show any significant difference after application of force. The activities of beta-glucuronidase increased significantly after application of orthodontic force and decreased with force deminished. These indicated that beta-glucuronidase was good indicator of bone remodelling resulted from initial orthodontic force.
Gingival Crevicular Fluid* ; Glucuronidase ; Humans ; Orthodontic Appliances

OBJECTIVES: The purpose of this study was to get help in order to diagnose orthopaedic disease, measure its activity and determine treatment plan by measuring the beta-glucuronidase activity in urine, serum and joint fluid. METHODS: The beta-glucuronidase activity was determined in the serum, urine and joint fluid of the patients with degenerative arthritis, rheumatoid arthritis, osteomyelitis and osteogenic sarcoma, and some other disease to study the change of the enzyme activity. These values of each specimen were calculated by standard curve and treated by statistical analysis. RESULTS: The results obtained were summarized as follows. 1. The beta-glucuronidase activity in the serum, urine and joint fluid was increased in patients with degenerative arthritis, rheumatoid arthritis, osteomyelitis and osteogenic sarcoma etc. 2. The increased beta-glucuronidase activity in the serum and joint fluid of each disease does not show a specific finding about respective disease, but the increased beta-glucuronidase activity was statistically significant in the urine of all disease groups(male:p=0. 0041, female:p=0. 0001). CONCLUSIONS: On the basis of these results, it was suggested that beta-glucuronidase activity was affected by the orthopaedic disease and differed according to each specimen.
Arthritis, Rheumatoid ; Glucuronidase* ; Humans ; Joints ; Osteoarthritis ; Osteomyelitis ; Osteosarcoma

No abstract available.
Breast* ; Glucuronidase* ; Infant, Newborn ; Jaundice, Neonatal* ; Milk* ; Milk, Human*

An efficient genetic transformation system and suitable promoters are essential prerequisites for gene expression studies and genetic engineering in streptomycetes. In this study, firstly, a genetic transformation system based on intergeneric conjugation was developed in Streptomyces rimosus M527, a bacterial strain which exhibits strong antagonistic activity against a broad range of plant-pathogenic fungi. Some experimental parameters involved in this procedure were optimized, including the conjugative media, ratio of donor to recipient, heat shock temperature, and incubation time of mixed culture. Under the optimal conditions, a maximal conjugation frequency of 3.05×10^-5 per recipient was obtained. Subsequently, based on the above developed and optimized transformation system, the synthetic promoters SPL-21 and SPL-57, a native promoter potrB, and a constitutive promoter permE^* commonly used for gene expression in streptomycetes were selected and their activity was analyzed using gusA as a reporter gene in S. rimosus M527. Among the four tested promoters, SPL-21 exhibited the strongest expression activity and gave rise to a 2.2-fold increase in β-glucuronidase (GUS) activity compared with the control promoter permE^*. Promoter SPL-57 showed activity comparable to that of permE^*. Promoter potrB, which showed the lowest activity, showed a 50% decrease in GUS activity compared with the control permE^*. The transformation system developed in this study and the tested promotors provide a basis for the further modification of S. rimosus M527.
Conjugation, Genetic ; Glucuronidase/genetics* ; Promoter Regions, Genetic ; Streptomyces rimosus/genetics*

OBJECTIVETo construct a lentiviral RNA interference system targeting heparanase (HPSE) based on miR30 and to test its silencing effect.

METHODSThree heparanase-shRNA structures were designed based miR30. The targeting fragments were obtained by PCR, then inserted into the vector LV PP-GFP to construct the recombinant lentiviral vector LV PP-GFP/miR-HPSE-shRNA, which was identified by PCR and sequencing. The 293T cells were co-transfect with LV PP-GFP/miR-HPSE-shRNA, pHelper 1.0 vector and pHelper 2.0 vector to produce lentiviruses, with which A375 cells were infected. Real-time fluorescence quantitative PCR and Western blot were performed to evaluate the expression of heparanase RNA and protein.

RESULTSThe lentiviral miR30-based RNAi vector targeting heparanase was constructed and confirmed by PCR and sequencing. The results of real-time fluorescence quantitative PCR and Western blot showed that the expression levels of both heparanase mRNA and protein in infected A375 cells were decreased significantly than those in control group.

CONCLUSIONThe lentiviral miR30-based RNAi vector targeting heparanase was been constructed successfully, which can be used for further study on RNAi-mediated oncolytic viruses.

Genetic Vectors ; Glucuronidase ; genetics ; Lentivirus ; genetics ; MicroRNAs ; genetics ; RNA Interference ; RNA, Small Interfering ; genetics