The objective of this study was to evaluate the relationship between beta-galactosidase activity in urine, in serum and urinary N-acetyl-beta-glucosarminidase activity as an early indicator of renal effect and mercurT concentration in urine and blood, reflecting the intensity of exposure to or the amount of body burden of mercury. This study was carried out among 70 workers exposed to mercury vapor and 63 non-exposed workers as a reference. The results were as follows ; 1. The mean concentration of urinary mercury (43.5 microgram/1) in exposed subjects was about nine times higher than that of non-exposed subjects, but the mean values of blood mercury were not different from each other 2. The mean values of beta-galactosidase activity in urine (119.7micromoleMU/h/g creatinine) and in blood (73.7 moIMU/H/l) of mercury-exposed subjects were significantly higher than those of non-exposed subjects. 3. In mercury-exposed subjects, beta-galactosidase activities in urine (r=0.38, p<0.01) and in serum (r=0.26, p<0.05) were correlated to urinary mercury concentration, but not to blood mercury concentration. The urinary excretion of beta-galactosidase activity was closely associated with urinary mercury concentration in the result of the multiple regression analysis. 4. The urinary beta-galactosidase activity in exposed subjects increased as the urinary mercury increased, and in the exposed subjects with more than 50microgram/1 of urinary mercury was highly related to urinary beta-galactosidase activity(r=0.47, p<0.05). 5. Among exposed subjects with more than 50microgram/l of urinary mercury, 20.0% of them showed abnormal value of urinary beta-galactosidase activity.
beta-Galactosidase* ; Body Burden

Genomic DNA and cDNA sequences of lactase from Aspergillus candidus were cloned. Sequences analysis revealed that the genomic DNA was 3458 bp containing eight introns, cDNA was 3015 bp and encoding a polypeptide of 1005 amino acid residues. Signal peptide was 19 amino acid residues, eleven potential N-glycosylation sites were assumed. Comparing the gene cDNA and amino acid sequences with other lactase sequences from various sources, it showed a very low homology with most of other sequences. Although the gene had a higher homology to Aspergillus oryzae lactase sequence, characterization of both enzymes exhibited obvious difference. The gene from Aspergillus candidus was a promising new gene for food industry.
Amino Acid Sequence ; Aspergillus ; enzymology ; genetics ; Cloning, Molecular ; Lactase ; Molecular Sequence Data ; Sequence Homology ; beta-Galactosidase ; chemistry ; genetics

OBJECT: The aim of this study is to determine whether exposure to chlorpromazine causes mutagenicity and genetic disorders. METHOD: Ames (Salmonella typhimurium) test and Rec assay (Bacillus subtilis) were used as indicators for DNA damage. Furthermore, the levels of umu operon expression by measuring the beta-galactosidase activity were monitered with the SOS umu test using S. typhimurium 1535 containing plasmid pSK1002. And the host-mediated assay was used to investigate the muta-genicity of chlorpromazine after the activation with in vivo metabolic systems. RESULTS: From the results, chlorpromazine did not affect DNA of S. typhimurium and B. subtilis strains and showed no mutagenicity at the all concentrations tested. These phenomena was also similar to that after metabolic activation of chlorpromazine in in vivo system. CONCLUSION: These results suggested that chlorpromazine did not show the mutagenicity and genotoxicity by four different methods used in this study.
beta-Galactosidase ; Biotransformation ; Chlorpromazine* ; DNA ; DNA Damage ; Operon ; Plasmids

ESWL is a safe and effective treatment for renal calculi. However, on destruction of the stone, the pressure of shock waves produces renal microdamage. Urinary enzyme testing has been used to diagnosis and monitor various types of renal injured. Three urinary enzymes, N-acetyl-beta- glucosaminidase, beta-galactosidase, gamma-glutamyl transferase in 24 hour urine were monitored in 20 patients before and after ESWL and in 5 controls. The ESWL, a single session treatment, was performed using EDAP LT-01. The amount of the shock wave treated was same regardless of stone size and location. The results were obtained as follows. 1. Post-ESWL N-acetyl-beta-glucosaminidase level was higher than that of control on day 1 and showed a rapid decrease on day 3, and returned to normal by day 28. 2. Post-ESWL beta-galactosidase level was higher than that of control on day 1 and was still higher on day 7, and returned to normal by day 28. 3. Post-ESWL gamma-glutamyl transferase level reached to a peak on day 1 and showed a rapid decrease on day 3, and returned to normal by day 28. In summary, ESWL for renal stones elevates urinary enzymes transiently, which was on peak one day after the treatment and returned to nearly normal level by 1 month after the treatment. Therefore, urinary enzymes seem to be useful in evaluation of the functional recovery of the kidney after ESWL.
beta-Galactosidase ; Diagnosis ; Hexosaminidases ; Humans ; Kidney ; Kidney Calculi ; Shock ; Transferases

The coeneal endothelium is essential for the maintenance of normal corneal hydration, thickness, and transparency. However, corneal endothelial cells are incapable of significant proliferation in vivo. As we age, the density of corneal endothelial (CEN) cells gradually decreases. The goal of our study is to explore the possibility of enhancing the proliferation of corneal endothelial cells by introduction of SV 40 large T antigen, a transforming protein. To this end, introduction of protein into CEN cells was assessed by liposome assisted beta-galactosidase transfection in vivo, ex vivo, and in vivo. In all cases, cells treated with liposome-protein complex have shown dramatic blue stain in beta-galactosidase activity staining. This result convinced us that we could artificially introduce a foreign protein into a cell. To ascertain where SV 40 large T antigen is localized in the cell, purified SV 40 large T antigen was transfected into the cells using liposome and its presence was determined immunohistochemically. We show that the liposome delivered SV 40 large is localized in the nucleus and mitotic figures which may suggest its functional activity.
Antigens, Viral, Tumor* ; beta-Galactosidase ; Endothelial Cells* ; Endothelium ; Liposomes ; Transfection*

OBJECTIVETo study the mechanism of lactose intolerance (LI) by cloning the mouse lactase cDNA and recombining a vector. METHODS Total murine RNA was isolated from the small intestine of a 4-week-old BALB/c mouse (d). Gene-specific primers were designed and synthesized according to the cDNA sequences of lactase-phlorizin hydrolase (LPH) in human, rat, and rabbit. A coding sequence (CDS) fragment was obtained using RT-PCR, and inserted into a clone vector pNEB-193, then the cDNA was sequenced and analyzed using bioinformatics.

RESULTSThe cDNA from the BALB/c mouse with 912 bp encoding 303 amino acid residues. Analysis of the deduced amino acid sequence using bioinformatics revealed that this cDNA shared extensive sequence homology with human LPH containing a conserved glycosyl hydrolase family 1 motif important for regulating lactase intolerance.

CONCLUSIONBALB/c mouse LPH cDNA (GenBank accession No: AY751548) provides a necessary foundation for study of the biological function and regulatory mechanism of the lactose intolerance in mice.

OBJECTIVES: This study was performed to evaluate the relationship between radiation induced senescence-like changes of endothelial cells and blood-brain barrier disruption. MATERIALS AND METHODS: Radiation of 15 Gy was applied to a monolayer culture of bovine aortic endothelial cells (BAEC). The morphological changes were observed over 19 weeks. An artificial blood-brain barrier (BBB) model was constructed using the Transwell(R) and co-culture of the BAEC with C6 glioma cells. After treatment with the same dose of radiation, changes in the BBB were observed by measurement of the trans-endothelial electrical resistance (TEER). RESULTS: Senescence-like changes of the endothelial cells appeared 1 week after irradiation; it was most prominent during the third week and replacement by normal endothelial cells was noted from the seventh week. The recovered normal endothelial monolayer was maintained until the 19th week. Senescence-like endothelial cells showed positive staining with senescence-associated beta-galactosidase (SA-beta-gal). In the Transwell(R), the TEER began to decrease 1 week after irradiation, and the decreased resistance reached its peak 18 days after irradiation, and then began to recover to some degree. CONCLUSION : After application of radiation (15 Gy), senescence-like changes of the endothelial cells were observed in the monolayer culture. These findings demonstrated good correlation with the disruption of blood-brain barrier in an in-vitro model of the BBB.
Aging ; beta-Galactosidase ; Blood-Brain Barrier ; Coculture Techniques ; Electric Impedance ; Endothelial Cells ; Glioma

The p53 tumor suppressor gene is one of the genes with greatest therapeutic potential for cancer treatment and its growth inhibitory mechanism is thought to be mediated through the activation of its downstream mediator, WAF1/Clip1. In this study, we evaluated the effect of the replication-defective recombinant adenovirus expressing wild-type p53 gene(Ad5CMV-p53) in human glioma cell lines(U-251, LG) harboring mutant-type p53. beta-galactosidase histochemistry revealed that 90% of the U-251 and 42% the of LG cells are infected with the adenovirus at a multiplicity of infection(MOI) of 25 plaque-forming units(PFU)/cell. Immunoblot analyses showed that endogenous p53 protein is expressed at a high level, and significant exogenous p53 protein expression and WAF1/Clip 1 induction peaked on day 1 and day 3 after Ad5CMV-p53 treatment. Introduction of Ad5CMV-p53 inhibited the cell rowth of U-251(85% inhibition) and LG cells(36% inhibition), and influenced cell morphology. The optimal dose of Ad5CMV-p53 for the tumor cel ls growth inhibition was MOI of 10-40 PFU/cell. These results suggest that Ad5CMV-p53 infects human glioma cells and transduces the p53 gene with high efficiency, and could be further developed for the gene therapy of human gliomas.
Adenoviridae ; beta-Galactosidase ; Brain* ; Genes, p53* ; Genes, Tumor Suppressor ; Genetic Therapy* ; Glioma* ; Humans*

OBJECTIVEThis study is to examine the secretion effects of beta-galactosidase in Lactococcus lactis.

METHODSThe usp45 and beta-galactosidase genes were cloned and inserted into plasmid pMG36e to obtain the recombinant plasmid pMG36e-usp-lacZ. This recombinant plasmid was transformed into both Escherichia coli DH5alpha and L. lactis MG1363. The enzyme activity, gene sequencing, SDS-PAGE and hereditary stability were assessed and studied.

RESULTSThe lacZ gene inserted into plasmids pMG36e-usp-lacZ was 99.37% similar to the GenBank sequence, and SDS-PAGE revealed an evident idio-strap at 116 KDa between L. lactis MG1363/pMG36e-usp-lacZ in both supernatant and cell samples. Beta-Galactosidase activity measured 0.225 U/mL in L. lactis pMG36e-usp-lacZ transformants, and its secretion rate was 10%. The plasmid pMG36e-usp-lacZ appeared more stable in MG1363.

CONCLUSIONThe authors concluded that these new recombinant bacteria well expressed and secreted beta-galactosidase, indicating that the beta-galactosidase expression system was successfully constructed, and this might provide a new solution for management of lactose intolerance specifically and promote the use of gene-modified organisms as part of the food-grade plasmid in general.

GM1 gangliosidosis is an autosomal recessive disorder caused by galactosidase beta1 (GLB1) gene variants which affect the activity of β-galactosidase (GLB). GLB dysfunction causes abnormalities in the degradation of GM1 and its accumulation in lysosome. This article reports the clinical and genetic features of a child with GM1 gangliosidosis. The girl, aged 2 years and 5 months, was referred to the hospital due to motor developmental regression for more than one year. Physical examination showed binocular deflection and horizontal nystagmus, but no abnormality was found on fundoscopy. The girl had increased muscular tone of the extremities, limitation of motion of the elbow, knee, and ankle joints, and hyperactive patellar tendon reflex. Blood biochemical examination showed a significant increase in aspartate aminotransferase. The 24-hour electroencephalographic monitoring detected frequent seizure attacks and diffuse θ wave activity, especially in the right hemisphere. Head magnetic resonance imaging showed thinner white matter in the periventricular region and diffuse high T2WI signal with unclear boundary. Three-dimensional reconstruction of white matter fiber tracts by diffusion tensor imaging showed smaller and thinner white matter fiber tracts, especially in the right hemisphere. Genetic analysis showed that the girl had compound heterozygous mutations of c.446C>T (p.Ser149Phe) and c.101T>C (p.Ile34Thr) in the GLB1 gene from her parents, among which c.101T>C (p.Ile34Thr) had not been reported in the literatures. The girl was finally diagnosed with GM1 gangliosidosis. Her conditions were not improved after antiepileptic treatment and rehabilitation training for 2 months.
Diffusion Tensor Imaging ; Female ; Gangliosidosis, GM1 ; genetics ; Humans ; Infant ; Mutation ; Virulence ; beta-Galactosidase ; genetics