• Proceedings of the SCSK Conference
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• 2003.09a
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• pp.285-290
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• 2003

Living cells are continuously subject to all sorts of stress such as ultraviolet rays on skin cells. Tests made in various laboratories show that when young fibroblasts (Le. at the beginning of their proliferate life) were repeatedly put under stress at subletal doses, they acquired a phenotype similar to Senescence Induced Prematurely by Stress (SIPS). The work presented hereafter was made on a new model of senescence induced prematurely by stress from ultraviolet Brays (UVB). The human fibroblast model was put under repeated UVB stress, causing SIPS. Several ageing biomarkers were used in order to characterise the cells that underwent stress:. an increase in the proportion of positive cells with senescence associated $\beta$-galactosidase activity (SA $\beta$-gal) measured by a specific coloration,. the proportion in the different morphological stages that fibroblasts undergo during culture visualised by microscopic observation,. the expression of genes known for overexpressing during senescence, particularly fibronectin and apolipoprotein J, measured by Real Time-PCR,. the common deletion of 4,977 bp in mitochondrial DNA, evaluated by nested PCR. Studying the variation of these 4 biomarkers, we have evaluated the protective effect of a Laminaria digitata extract (LDE) that can be used as a natural active ingredient for anti-ageing cosmetics.

• KSBB Journal
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• v.10 no.3
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• pp.317-322
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• 1995

Insect cells were grown and infected with baculovirus for the production of recombinant protein. Later infection gave the lower expression of recombinant protein. This indicates that the expression rate is lower at higher cell concentration. This phenomena provides a well-posed optimization problem with respect to the infection time. The optimal infection time was experimentally shown to exist for the maximum productivity of recombinant protein. Also, the expression increased with the addition of 5% silkworm hemolymph. This is considered to be due to the increase of intracellular viruses and the longevity of viable cells after the infection. The production of ${\beta}$-galaclosidase increased about ten-fold with the addition of yeastolate and silkworm hemolymph for high cell density and high expression, respectively.

• BMB Reports
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• v.28 no.1
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• pp.21-26
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• 1995

The Escherichia coli mixed-function serC-aroA operon encodes biosynthethic enzymes for unrelated pathways leading to the syntheses of serine and aromatic amino acids. It has been proposed that the operon is expressed in a cAMP-dependent manner. In this work experiments were performed to investigate the cAMP-dependent expression of the operon. Exogenous cAMP increased ${\beta}$-galactosidase synthesis in the $cya^+$ and cya strains harboring the serC-aroA-lac fusion plasmid. This enhancement was more dramatic in the $cya^-$ strain grown in a minimal medium. In a dot blot assay the serC-aroA mRNA content increased in a concentration-dependent pattern after the addition of exogenous cAMP. The activity of phosphoserine aminotransferase, encoded by the serC gene, apparently increased in E. coli cells after the addition of cAMP. All results obtained confirmed that the expression of the E. coli serC-aroA operon is positively regulated by cAMP at the level of transcription.

• The Journal of Korean Physical Therapy
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• v.23 no.5
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• pp.35-41
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• 2011

Purpose: This study was carried out to investigate the spatiotemporal localization of the amygdaloid nucleus innervating the rat stomach using PRV-BaBlu, which has been known to be an excellent type of neurotracer with the ability to transpass the neuronalsynaptic cleft. Methods: Ninety Sprague-Dawley rats (250~300 g) that were injected with PRV-BaBlu into the stomach were randomly divided into 3, 4 and 5 day groups (each group n=30). $2{\mu}l$ of PRV-BaBlu, a genetically modified strain of PRV-Bartha with the lac-Z gene,was injected into the rat stomach and immunostained with a mouse anti-${\beta}$-galactosidase at 3, 4 and 5 days after the virus injection. Results: The PRV-BaBlu infected the neurons in the amygdaloid nucleus, and the degree of viral infection in experimental animals showed a tendency to increase significantly with time (p<0.05). The neurons between the left and right amygdaloid nucleus significantly differ (p<0.05). Conclusion: This showed that PRV-BaBlu was an excellent neurotracer for localizing the amygdaloid nucleus, and the amygdaloid nucleus has a sensory input and motor output on stomach movement, influencing emotional behavior.

• Biotechnology and Bioprocess Engineering:BBE
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• v.5 no.4
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• pp.288-305
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• 2000

A review on the main aspects associated with yeast flocculation and its application in biotechnological processes is presented. This subject is addressed following three main aspects-the basics of yeast flocculation, the development of "new" flocculating yeast strains and bioreactor development. In what concerns the basics of yeast flocculation, the state of the art on the most relevant aspects of mechanism, physiology and genetics of yeast flocculation is reported. The construction of flocculating yeast strains includes not only the recombinant constitutive flocculent brewer's yeast, but also recombinant flocculent yeast for lactose metabolisation and ethanol production. Furthermore, recent work on the heterologous $\beta$-galactosidase production using a recombinant flocculent Saccharomyces cerevisiae is considered. As bioreactors using flocculating yeast cells have particular properties, mainly associated with a high solid phase hold-up, a section dedicated to its operation is presented. Aspects such as bioreactor productivity and culture stability as well as bioreactor hydrodynamics and mass transfer properties of flocculating cell cultures are considered. Finally, the paper concludes describing some of the applications of high cell density flocculating bioreactors and discussing potential new uses of these systems.e systems.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.12
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• pp.1808-1812
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• 2002

The present study was carried out to examine the efficiency of microcapsules and a stability of lactase in vitro in the simulated gastric and intestinal conditions. As a coating materials, medium-chain triacylglycerol (MCT) and polyglycerol monostearate (PGMS) were used. The highest efficiency of microencapsulation was found in the ratio of 15:1 as coating to core material with both MCT (91.5%) and PGMS (75.4%). In a subsequent experiment, lactose content was measured to study a microcapsule stability. Lysis of microcapsules made by MCT in simulated gastric fluid was proportionally increased such as 3% in pH 5 and 11% in pH 2 for 20 min incubation. In the case of PGMS microcapsulation, 11-13% of lactose was hydrolyzed at 20 min in all pHs and also very little amount (less than 3%) of lactose was hydrolyzed after 20 min in all pHs. The highest percentages of lactose hydrolysis in MCT and PGMS microcapsules were 68.8 and 60.8% in pHs 7 and 8 during 60 min, respectively. Based on our data, the lactase microcapsules seemed to be stable when they stay in the stomach, and hydrolyzed rapidly in small intestine where the bile acid was excreted.

• BMB Reports
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• v.36 no.4
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• pp.379-386
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• 2003

A mutant herpes simplex virus 1, mtHSV, was constructed by inserting the E. coli beta-galactosidase gene into the loci of icp34.5, the apoptosis-inhibiting gene of HSV. The mtHSV replicated in and lysed U251 (human glioma cells), EJ (human bladder cells), and S-180 (mice sarcoma cells), but not Wish (human amnion cells) cells. With its intact tk (thymidine kinase) gene, mtHSV exhibited susceptibility to acyclovir (ACV), which provided an approach to control viral replication. An in vivo test with mtHSV was conducted in immune-competent mice bearing sarcoma S-180 tumors, which were treated with a single intratumoral injection of mtHSV or PBS. Tumor dimensions then were measured at serial time points, and the tumor volumes were calculated. Sarcoma growth was significantly inhibited with prolonged time and reduced tumor volume. There was microscopic evidence of necrosis of tumors in treated mice, whereas no damage was found in other organs. Immunohistochemical staining revealed that virus replication was exclusively confined to the treated tumor cells. HSV-1 DNA was detected in tumors, but not in the other organs by a polymerase chain reaction analysis. From these experiments, we concluded that mtHSV should be a safe and promising oncolytic agent for cancer treatment.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.5
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• pp.705-711
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• 2004

Growth promoters were added to skim milk to retain the viability of Lactobacillus acidophilus and Bifidobacterium longum to help the product meet the "therapeutic minimum" at the time of consumption. The experiments were divided into two parts. The first part of the study used chicory inulin, isomalto-oligosaccharides and sucrose to investigate the effects of sugars on the activity of L. acidophilus and B. longum. The results indicated that the addition of isomalto-oligosaccharides stimulated growth of L. acidophilus and B. longum, resulting in a higher level of the probiotics after one month storage and yielded better $\beta$-galactosidase activity during fermentation. The second part studied the effects of three growth promoters on the viability of the probiotic cultures and the response surface method was employed to find the optimal ratio for addition of the growth promoters. The optimal ratio for added calcium gluconate, sodium gluconate and N-acetylglucosamine in fermented milk drinks were established. The response surface method proved to be a very effective way of optimizing the activity of probiotic cultures when developing a new fermented milk drink.

• Journal of mucopolysaccharidosis and rare diseases
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• v.4 no.1
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• pp.21-25
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• 2018

Fabry disease is a hereditary lysosomal storage disorder caused by the reduction or absence of lysosomal enzyme alpha-galactosidase A and the accumulation of glycosphingolipids, such as globotriaosylceramide (Gb3), in various organs, including the heart. The prevention of cardiac involvement in Fabry disease can only be achieved by enzyme replacement therapy (ERT), and the method of assessing the efficacy of ERT should be confirmed. Changes in the electrocardiogram, such as the shortening of PQ interval, prolongation of QTc and repolarization abnormalities as well as left ventricular hypertrophy in voltage criteria, can be used to identify Fabry disease patients; however, the usefulness of electrocardiograms for evaluating the efficacy of ERT is limited. The assessment of left ventricular hypertrophy using echocardiography has been established to evaluate the efficacy of ERT during long-term period. A new technique involving speckled tracking method might be useful for detecting early cardiac dysfunction and identifying the effect of ERT for a relatively short period. The estimation of left ventricular hypertrophy using cardiac magnetic resonance (CMR) is also useful for assessing the efficacy of ERT. Identifying late gadolinium enhancement in CMR may affect the effectiveness of ERT, and the new technique of T1 mapping might be useful for monitoring the accumulation of Gb3 during ERT. Histopathology in cardiac biopsy specimens is another potentially useful method for identifying the accumulation of GB3; however, the use of histopathology to evaluate of the efficacy of ERT is limited because of the invasive nature of an endomyocardial biopsy.

• Reproductive and Developmental Biology
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• v.35 no.2
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• pp.185-190
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• 2011

To evaluate the effect of spermatozoa culture on glycosidase activity of frozen-thawed spermatozoa in human, the spermatozoa were treated experimentally and assayed for activities of ${\alpha}$-L-fucosidase, ${\alpha}$-D-mannosidase, ${\beta}$-D-galactosidase and N-acetyl-${\beta}$-D-glucosaminidase (${\beta}$-GlcNAc'ase). The ${\beta}$-GlcNAc'ase activity was at least two-folds higher than other glycosidases regardless of spermatozoa incubation (p<0.05). The spermatozoa motility was decreased with incubation periods, but no effects by different glycosidases on the changes of spermatozoa motility during the various periods of incubation. In all glycosidases, the spermatozoa-zona binding rates in spermatozoa without incubation were higher than in spermatozoa incubated for 2 h (p<0.05). ${\beta}$-GlcNAc'ase is present mainly in the plasma membrane of spermatozoa frozen-thawed in human. It was also shown that the glycosidase activity was increased in all glycosidases in spite of lower sperm-zona binding by spermatozoa incubation.