• Proceedings of the Korean Society of Applied Pharmacology
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• 1995.04a
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• pp.89-89
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• 1995

Surface epithelial cells isolated from hamster tracheas and grown on a thick collagen gel become a highly enriched population of mucus-secreting cells. Epithelial cells from tracheas of hamsters were collected using enzymatic procedures and cultured under various conditions. The medium used consisted of a 1:1 mixture of medium 199 and Dulbecco's modified Eagle's (DME) medium which was conditioned before use. Insulin, transferrin, hydrocortisone, epidermal growth factor, and extract from bovine hypothalamus were used as supplement. Due to relatively low basal rates of min secretion from in vitro cultures, cultures are generally radiolabeled using $^3$H-glucosamine as a metabolic precursor. The radiolabeled mucinsreleased are quantitated by precipitation with TCA/PTA. Using this cell culture system, we investigated mucin release of goblet cells by altering the media bathing the apical surface of hamster tracheal surface epithelial(HTSE) cells. Acidic media added sulfuric acid caused sigcificant increases in mucin relesse (155${\pm}$20% at pH 4 and 146${\pm}$16% at, pH 5). Ammonium hydroxide also increased mucin release at pH 9.0(156${\pm}$17%) and pH 10(295${\pm}$9%) respectively. This additional mucin release seems to be associated with cell membrane damage as indicated by release of cellular LDH. SP stimulates secretion of mucin in cultured HTSE cells(154${\pm}$16% at 1${\times}$10$\^$-6/M and 165${\pm}$25% at 1${\times}$10$\^$-5/M. PAF at 5${\times}$10$\^$-6/M and 5${\times}$10$\^$-5/M enhanced by HTSE cells in vitro 168${\pm}$34% and 259${\pm}$30% of mucin secretion, respectively. The increase in mucin release by PAF and SP was not secondary to cell damage or necrosis. SP and PAF may be in mediating mucous secretion induced by inflammation irritantion and infection.

• Journal of Microbiology
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• v.45 no.1
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• pp.21-28
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• 2007

In order to search for specific genotypes related to this unique phenotype, we used whole genomic DNA microarray to characterize the genomic diversity of Helicobacter pylori (H. pylori) strains isolated from clinical patients in China. The open reading frame (ORF) fragments on our microarray were generated by PCR using gene-specific primers. Genomic DNA of H. pylori 26695 and J99 were used as templates. Thirty-four H. pylori isolates were obtained from patients in Shanghai. Results were judged based on In(x) transformed and normalized Cy3/Cy5 ratios. Our microarray included 1882 DNA fragments corresponding to 1636 ORFs of both sequenced H. pylori strains. Cluster analysis, revealed two diverse regions in the H. pylori genome that were not present in other isolates. Among the 1636 genes, 1091 (66.7%) were common to all H. pylori strains, representing the functional core of the genome. Most of the genes found in the H. pylori functional core were responsible for metabolism, cellular processes, transcription and biosynthesis of amino acids, functions that are essential to H. pylori's growth and colonization in its host. In contrast, 522 (31.9%) genes were strain-specific genes that were missing from at least one strain of H. pylori. Strain-specific genes primarily included restriction modification system components, transposase genes, hypothetical proteins and outer membrane proteins. These strain-specific genes may aid the bacteria under specific circumstances during their long-term infection in genetically diverse hosts. Our results suggest 34 H. pylori clinical strains have extensive genomic diversity. Core genes and strain-specific genes both play essential roles in H. pylori propagation and pathogenesis. Our microarray experiment may help select relatively significant genes for further research on the pathogenicity of H. pylori and development of a vaccine for H. pylori.

• Development and Reproduction
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• v.3 no.1
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• pp.29-38
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• 1999

The pathogenesis of endometriosis is unknown, but retrograde menstruation is widely accepted as an etiology. Refluxed endometrium from endometriosis patients is more prone to implant and invade peritoneum possibly through the action of extracellular proteolysis. This proteolytic action may involve plasminogen activators and the collagenase system. Plasminogen activators (PAs) and matrix metalloproteinases (MMPs) play a critical role in the breakdown of extracellular matrix components and basement membrane in the processes of implantation and tumor invasion. PAs are inhibited by plasminogen activator inhibitor (PAI) and MMPs activity is inhibited by tissue inhibitor of metalloproteinase (TIMP). To test the hypothesis that lower expression of PAI-1 and TIMP-3 in endometrium from women with endometriosis, we investigated their PAI-1 and TIMP-3 expression by quantitative competitive RT PCR in endometrium from women with and without endometriosis. Endometrial tissues were obtained from 14 patients with severe endometriosis and 14 patients without endometriosis. Total RNA was extracted and reverse transcribed into cDNA, and quantitative competitive PCR (QC PCR) was performed to evaluate PAI-1 and TIMP-3 mRNA expression. Endometrium from patients with endometriosis showed decreased expression of PAI-1 and TIMP-3 mRNA compared to endometrium from control in luteal phase (p<0.05). Our results suggest that endometrium from women with endometriosis expresses lower levels of PAI-1 and TIMP-3 than endometrium from normal women. Endometrium from endometriosis patients may be more invasive and prone to peritoneal implantation than control because of higher PA and MMP enzymatic activity. Thus, increased proteolytic activity may be one of the reasons for the invasive properties of the endometrium resulting in the development of endometriosis.

• Journal of Biomedical Engineering Research
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• v.25 no.5
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• pp.329-334
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• 2004

The electronic nose (E-nose) recently finds its applications in medical diagnosis, specifically on detection of diabetes, pulmonary or gastrointestinal problem, or infections by examining odors in the breath or tissues with its odor characterizing ability. The odor recognition performance of E-nose can be improved by manipulating the sensor array responses of vapors in time-profile forms. The different chemical interactions between the sensor materials and the volatile organic compounds (VOC's) leave unique marks in the signal profiles giving more information than collection of the conventional piecemal features, i.e., maximum sensitivity, signal slopes, rising time. In this study, to use them in vapor recognition task conveniently, a novel time-profile method was proposed, which is adopted from digital image pattern matching. The degrees of matching between 8 different vapors were evaluated by using the proposed method. The test vapors are measured by the silicon-based gas sensor array with 16 CB-polymer composites installed in membrane structure. The results by the proposed method showed clear discrimination of vapor species than by the conventional method.

• Journal of Life Science
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• v.27 no.11
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• pp.1340-1348
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• 2017

Sanguinarine is a benzophenanthridine alkaloid derived from the roots of Sanguinaria canadensis L., which is used for the purpose of treating various diseases. Although studies of anticancer activities have been performed using various cancer cell lines, the phenomenon of inducing apoptosis in cancer cells by using sanguinarine requires more research. Therefore, this study investigated the anti-cancer activities and related mechanisms of sanguinarine used with Hep3B human hepatocellular carcinoma cells in terms of the regulation of apoptosis. Sanguinarine inhibited the proliferation of Hep3B cells in a concentration-dependent manner, which was associated with the induction of apoptosis. Sanguinarine also increased the activity of caspase-3, which is a typical effector caspase, and the activities of caspase-8 and caspase-9, which are key when initiating extrinsic and intrinsic apoptosis pathways, respectively. In addition, sanguinarine increased the expression of death receptor-related genes and pro-apoptotic BAX, which belongs to the Bcl-2 family, while suppressing the expression of anti-apoptotic Bcl-2. Sanguinarine promoted the truncation of Bid and enhanced the release of cytochrome c from the mitochondria to the cytoplasm due to a loss of mitochondrial membrane potential. Furthermore, the reduction of a survival rate that was induced by sanguinarine and the induction of apoptosis disappeared with the inhibition of artificial caspase activity. Therefore, the results of the study indicated that sanguinarine-induced apoptosis in Hep3B cells involves both extrinsic and intrinsic pathways; such apoptosis is a caspase-dependent phenomenon.

• Journal of Life Science
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• v.28 no.5
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• pp.627-637
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• 2018

Motility and chemotaxis are crucial for disease development in many motile pathogens, including spirochetes. In many bacteria, motility is provided by flagella rotation, which is controlled by a chemotaxis-signal-transduction system. Thus, motility and chemotaxis are inextricably linked. Spirochetes are a unique group of bacteria with distinctive flat-wave morphology and corkscrew-like locomotion. This unusual motility pattern is believed to be important for efficient motility within the dense tissues through which these spirochetes preferentially disseminate in a host. Unlike other externally flagellated bacteria-where flagella are in the ambient environment-the flagella of spirochetes are enclosed by the outer membrane and thus are called periplasmic flagella or endoflagella. Although motilityand chemotaxis-associated genes are well studied in some bacteria, the knowledge of how the spirochete achieves complex swimming and the roles of most of the putative spirochetal chemotaxis proteins are still elusive. Recently, cutting-edge imaging methods and unique genetic manipulations in spirochetes have helped to unravel the mystery of motility and chemotaxis in spirochetes. These contemporary advances in understanding the motility and chemotaxis of spirochetes in a host's persistence and disease process are highlighted in this review.

• Korean Journal of Food Science and Technology
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• v.31 no.2
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• pp.410-415
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• 1999

Yakju was sterilized with high-voltage pulses of short time of a continuous pulsed electric field (PEF) system. The initial microbial counts of Yakju were $2.2{\times}10^{5}$ CFU/mL for total aerobes. The pH, acidity and electric conductivity of Yakju were 3.82, 0.37% and 1.24 mS/cm, respectively. Yakju was treated with exponential-wave formed electric pulses of 100 Hz for $0{\sim}4000{\mu}s$ under the field strength of $20{\sim}35\;kV/cm$. The lethal effect of electric fields on microorganisms was resulted from the breakdown of the cell membrane induced by the transmembrane electric potential. The critical values of the external field for the sterilization were 16.0 kV/cm for total aerobes. Logarithmic survival rates decreased linearly at low electric field strength, but curvilinearly at high electric field strength with treatment time. The sterilization of Yakju was more largely affected by the electric field strength than by the treatment time. Any changes in pH, acidity, and the growth of microorganisms were not found in the PEF treated Yakju during the storage at both $4^{\circ}C\;and\;30^{\circ}C$.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.41 no.3
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• pp.243-252
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• 2015

Melanosome, the pigment granule in melanocyte, determines the color of skin when it moves into the keratinocyte. Inhibition of melanosome transfer from melanocyte to keratinocyte results in skin depigmentation. Protease activated receptor-2 (PAR-2) is involved in signal transduction systems via cell membrane and increases the melasome transfer when it is activated by cleavage of their extracellular amino acid sequence by trypsin or by a peptide such as SLIGKV. Here, we showed that lobaric acid inhibited PAR-2 activation and affected the mobilization of $Ca2^+$. The uptake of fluorescent microspheres and isolated melanosomes from melan-a melanocytes to keratinocytes induced by SLIGKV were inhibited by lobaric acid. Also, confocal microscopy studies illustrated a decreased melanosome transfer to keratinocytes in melanocyte-keratinocyte co-culture system by lobaric acid. In addition, lobaric acid induced visible skin lightening effect in human skin tissue culture model, melanoderm$^{(R)}$. Our data suggest that lobaric acid could be an effective skin lightening agent that works via regulation of phagocytic activity of keratinocytes.

• Food Science and Preservation
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• v.11 no.4
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• pp.530-537
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• 2004

This study was conducted to determine the inactivation effect of electrolyzed water and organic acids either alone or in combination on L. monocytogenes or natural microflora on lettuce. Acidic electrolyzed water completely inactivated L. monocytogenes in broth system within 60 sec, but alkalin electrolyzed water caused approximate 1.7 log CFU/g reduction. However, acidic electrolyzed water reduced only 2.5 log CFU/g of L. monocytogenes on lettuce, and similar antimicrobial effect was observed with alkalin electrolyzed water. In the meantime, acidic and alkaline electrolyzed water caused approximately 2 log CFU/g reduction compared to control, whereas both electrolyzed water combined with $1\%$ organic acids ranged from 2.6 to 3.7 log CFU/g reduction. Among the organic acids, both electrolyzed water combined with $1\%$ citric acid showed the strongest synergistic antimicrobial effect to reduce L. monocytogenes on lettuce as well as total counts, yeast and molds. When antimicrobials, alone or in combination were treated into L. monocytogenes inoculated lettuce at $5^{\circ}C\;and\;15^{\circ}C$ for designed periods, the combined alkalin electrolyzed water with $1\%$ citric acid showed the greatest potential to inhibit growth of the bacteria. According to Scanning Electron Microscopy(SEM), the treatment of electrolyzed alkali water in combination with $1\%$ citric acid highly reduced the growth of the L. monocytogenes compared to single treatment and resulted in causing the destruction of cell membrane.

• Animal cells and systems
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• v.12 no.4
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• pp.313-319
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• 2008

The ultrastructure of oocytes during oogenesis and oocyte degeneration associated with follicle cells in female Sinonovacula constricta(Lamarck, 1818) were investigated by electron microscope observations. Ovarian follicles are surrounded by a matrix of vesicular connective tissue cells(VCT cells). VCT cells contain large quantities of glycogen particles and several lipid droplets in their cytoplasm. It is suggested that VCT cells act as a source of nutrients for vitellogenesis during oogenesis. In early vitellogenic oocytes, several coated vesicles, which appear at the basal region of the oocyte, lead to the formation of membrane-bound vesicles via endocytosis. The uptake of nutritive materials in coated vesicles formed by endocytosis appears through the formation of coated pits on the oolemma during vitellogenesis. During the late stage of oogenesis, yolk precursors(yolk granules), mitochondria and lipid droplets are present in the cytoplasm of late vitellogenic oocytes. In particular, proteinaceous yolk granules containing several different components are intermingles and form immature yolk granules. In the mature oocyte, small immature yolk granules are intermingled and form large mature yolk granules. Vitellogenesis occurs through a process of autosynthesis, involving combined activity of the Golgi complex, mitochondria and rough endoplasmic reticulum in the cytoplasm of vitellogenic oocytes. The process of heterosynthesis is where extraovarian precursors are incorporated into oocytes by endocytosis at the basal region of early vitellogenic oocytes before the formation of the vitelline coat. Follicle cells appear to play an important role in vitellogenesis and oocyte degeneration. The functions of attached follicle cells to the oocyte during oocyte degeneration are phagocytosis and digestion of phagosomes originating from oocyte degeneration. After digestion of phagosomes, it is assumed that the function of follicle cells can permit a transfer of yolk precursors necessary for vitellogenesis and allows for the accumulation of glycogen and lipid during oocyte degeneration, which can be employed by vitellogenic oocytes. Follicle cells of S. constricta may possess a lysosomal system for induction of oocyte breakdown and might resorb phagosomes in the cytoplasm for nutrient accumulation during oocyte degeneration.