• Parasites, Hosts and Diseases
• /
• 제40권2호
• /
• pp.93-99
• /
• 2002

The effect of a secretory proteinase from the pathogenic amoebae Acanthamoeba castellanii on host's defense-oriented or regulatory proteins such as immunoglobulins, interleukin-1, and protease inhibitors was investigated. The enzyme was found to degrade secretory immunoglobulin A (slgA), IgG, and IgM. It also degraded $interleukin-1{\alpha}$ ($IL-l{\alpha}$) and $IL-l{\beta}$. Its activity was not inhibited by endogenous protease inhibitors, such as ${\alpha}$2-macroglobulin, ${\alpha}l-trypsin$ inhibitor, and ${\alpha}2-antiplasmin$. Furthermore, the enzyme rapidly degraded those endogenous protease inhibitors as well. The degradation of host's defense-oriented or regulatory proteins by the Acanthanoeba proteinase suggested that the enzyme might be an important virulence factor in the pathogenesis of Acanthamoeba infection.

• 대한기관식도과학회:학술대회논문집
• /
• 대한기관식도과학회 1993년도 제27차 학술대회 초록집
• /
• pp.67-67
• /
• 1993

기관 및 후두의 점막표면은 중이나 이관과 마찬가지로 점막섬모 정화작용, 면역글로린, 항세균 효소 등의 다양한 점막 방어기전에 의해 보호되고 있는데 기관 및 후두의 분비선과 분비세포의 형태에 관한 보고는 많으나 분비활성도와 연관된 분비조직의 발생에 대한 연구는 충분치 못하다. 저자들은 백서 기관 및 후두의 분비조직의 발생을 알아보고 향후 기관 및 후두의 발생형태학적 연구의 기초자료로 삼기 위해서 임신 16일부터 생후 21사이의 백서를 이용하여 H & E 염색, AB-PAS 염색과 lysozyme의 면역조직화학적 방법을 통하여 백서 기관 및 후두분비조직의 발달을 연구하였다. 그 결과 백서 기관 및 후두의 분비기능은 출생후 폐의 통기와 함께 활성화되는 것으로 해석되었다.

• Journal of Microbiology and Biotechnology
• /
• 제21권4호
• /
• pp.412-420
• /
• 2011

Secretion of cellobiase occurred in a brefeldin A (BFA) uninhibited manner in the filamentous fungus Termitomyces clypeatus. Fluorescence confocal microscopy revealed that application of the drug at a concentration of 50 ${\mu}g$/ml caused arrest of Spitzenkorper assembly at the hyphal tip. This resulted in greater than 30% inhibition of total protein secretion in the culture medium. However, the cellobiase titer increased by 17%, and an additional 13% was localized in the vacuolar fraction en route secretion. The secretory vacuoles formed in the presence of the drug were also found to be bigger (68 nm) than those in the control cultures (40 nm). The enzyme secreted in the presence and absence of BFA revealed a single activity band in both cases in native PAGE and had similar molecular masses (approx. 120 kDa) in SDS-PAGE. The BFA enzyme retained 72% of native glycosylation. It also exhibited a higher stability and retained 98% activity at $50^{\circ}C$, 93.3% activity at pH 9, 63.64% activity in the presence of 1M guanidium hydrochloride, and 50% activity at a glucose concentration of 10 mg/ml in comparison to 68% activity, 75% activity, 36% activity, and 19% activity for the control enzyme, respectively. The observations collectively aimed at the operation of an alternative secretory pathway, distinct from the target of brefeldin A, which bypassed the Golgi apparatus, but still was able to deliver the cargo to the vacuoles for secretion. This can be utilized in selectively enhancing the yield and stability of glycosidases for a successful industrial recipe.

• BMB Reports
• /
• 제30권6호
• /
• pp.379-384
• /
• 1997

The effects of several ions on the specific activity and isozyme patterns of cellular and secretory isoperoxidases were studied in suspension-cultured cells of rice (Oryza sativa L.). Peroxidase release into the culture medium occurred in the absence of added calcium. The addition of calcium ion greatly stimulated the secretion of cationic isoperoxidases such as C2 and C3 into the medium: a maximum 11 fold increase of secretions occurred in the presence of 5 mM $CaCl_2$, and the secretion was accomplished within 1 hour after the addition of $CaCl_2$. About a 10 fold increase of the peroxidase secretion into the medium did occur with 0. 5% NaCl, whereas cellular isoperoxidase levels were reduced notably. About a 6 fold increase of the specific activity of cellular isoperoxidase was found in 5 mM $NiCl_2$-treated cell, while $NiCl_2$ had no effect on the secretion of peroxidase into the medium. Various concentrations of KCl did not change peroxidase secretion, but 5 mM $ZnCl_2$ reduced peroxidase secretion greatly. The major secretory isoperoxidases stimulated by $CaCl_2$, NaCl and cellulase were composed of cationic isoperoxidases C2 and C3, which were found to be localized in the cell wall of rice by examination of the enzyme in the protoplast. Furthermore, the secretion rates of secretory isoperoxidases were increased rapidly when cellulase was treated in the absence of the osmotic stabilizer of 0.4 M mannitol. These results suggest that the stimulations of secretory isoperoxidase levels seem to be due to the stimulation of secretion into the culture medium of rice.

• Restorative Dentistry and Endodontics
• /
• 제19권2호
• /
• pp.372-383
• /
• 1994

Saliva plays an important role in modulating the oral microbial ecology. And it is suggested to influence the initiation and progression of the dental caries. To evaluate the correlations between the salivary antimicrobial agents and the caries susceptibility, the 51 subjects were divided into 3 groups according to caries experience ; caries resistant group, medium caries susceptible group, and high caries susceptible group. Stimulated whole saliva was collected, and the salivary levels were measured for lysozyme, lactoferrin, and secretory-IgA to Streptococcus mutans. The lysozyme level was estimated using Micrococcus diffusion plate, lactoferrin level was determined with a non-competitive avidin-biotin enzyme immunoassay, and the titer of secretory IgA to Streptococcus mutans was assayed with ELISA. The results were as follows: 1. Lysozyme levels of each group showed no significant difference statistically (p>0.05). 2. The caries resistant group and the medium caries susceptible group had significantly higher levels of lactoferrin than the high caries susceptible group (p<0.05). But no clear difference was observed between the caries resistant group and the medium caries susceptible group(p>0.05). 3. The caries resistant group and the medium caries susceptible group showed relatively higher levels of the secretory IgA to Streptococcus mutans than the pigh caries susceptible group, but no significant difference was observed statistically (p>0.05).

• Parasites, Hosts and Diseases
• /
• 제41권3호
• /
• pp.165-169
• /
• 2003

This study describes the characterization of 80 kDa pretense showing gelationlytic property among three pretenses in the excretory/secretory proteins (ESP) from Toxoplasma gondii. The pretense activity was detected in the ESP but not in the somatic extract of RH tachyzoites. This pretense was active only in the presence of calcium ion but not other divalent cationic ions such as $Cu^{2+},{\;}Zn^{2+},{\;}Mg^{2+},{\;}and{\;}$Mn^{2+}$, implying that $Ca^{2+}$ is critical factor for the activation of the protease. The 80 kDa pretense was optimally active at pH 7.5. Its gelatinolytic activity was maximal at $37^{\circ}C$, and significant level of enzyme activity of the pretense remained after heat treatment at $56^{\circ}C$ for 30 min or $100^{\circ}C$ for 10 min, This thermostable enzyme was strongly inhibited by metal chelators, i.e., EDTA, EGTA, and 11 10-phenanthroline. Thus, the 80 kDa pretense in the ESP secreted by T. gondii was classified as a calcium dependent neutral metalloprotease.

• 대한약리학회지
• /
• 제30권2호
• /
• pp.205-215
• /
• 1994

It is well known that chronic stimulation with CCK gives rise to growth of exocrine pancreas and to increased content of enzyme proteins in pancreas. However, littls Is known about changes of the secretory function of exocrine pancreas which has been chronically stimulated with CCK, especially about the responsiveness to secretagogues such as CCK, caerulein and carbachol. The present study was performed to investigate the effect of camostat on secretory profiles and the responsiveness to secretagogues of exocrine pancreas by observing in vitro amylase release stimulated by cholecystokinin-octapeptide(CCK-8) and carbachol in dispersed isolated pancreatic acini from camostat-treated rats for 4 or 10 days. The results summarized as follows : 1) The maximal effective concentration of CCK-8 in amylase release in the camostat treated group was greater than control group, but that of carbachol was not different between groups. 2) Analysis of the stimulated amylase release as the percentage of the maximal response revealed that camostat treatment caused right-shift of the dose-response curve of CCK-8. Camostat did not cause significant changes in the dose-response curve of carbachol. 3) There were considerable increases in the amylase release in the camostat-treated group, compared to the control when acini were stimulated with CCK-8 $10^{-9}\;M$ and carbaochol $10^{-6}\;M$, and higher concentrations. 4) There was a reverse correlation between the tissue content and the maximal release(percent of the total content) of amylase. These results suggest that chronic exposure of exocrine pancreas to increased endogenous CCK can enhance the responsiveness of exocrine enzyme secretion to secretagogues, especially at higher concentrations of CCK and carbachol.

• Journal of Microbiology and Biotechnology
• /
• 제24권2호
• /
• pp.197-208
• /
• 2014

A novel protease gene from Bacillus gibsonii, aprBG, was cloned, expressed in B. subtilis, and characterized. High-level expression of aprBG was achieved in the recombinant strain when a junction was present between the promoter and the target gene. The purified recombinant enzyme exhibited similar N-terminal sequences and catalytic properties to the native enzyme, including high affinity and hydrolytic efficiency toward various substrates and a superior performance when exposed to various metal ions, surfactants, oxidants, and commercial detergents. AprBG was remarkably stable in 50% organic solvents and retained 100% activity and stability in 0-4 M NaCl, which is better than the characteristics of previously reported proteases. AprBG was most closely related to the high-alkaline proteases of the subtilisin family with a 57-68% identity. The secretion and maturation mechanism of AprBG was dependent on the enzyme activity, as analyzed by site-directed mutagenesis. Thus, when taken together, the results revealed that the halo-solvent-tolerant protease AprBG displays significant activity and stability under various extreme conditions, indicating its potential for use in many biotechnology applications.

• 한국치위생학회지
• /
• 제16권3호
• /
• pp.463-469
• /
• 2016

Objectives: The purpose of the study is to investigate the effect of chewing gum containing Prunus mume extract(PME) on the change of saliva ingredients. On the basis of the biological background of molecules and diagnostic indices in the use of saliva, the mastication effect of chewing gum containing PME was demonstrated in terms of secretory IgA concentration and total protein concentration in stimulated saliva. Methods: This study is an experimental research on the use of a research design before and after applying a randomized control group. Participants were distributed randomly to the experimental group and the control group, respectively. The experiment group was instructed to masticate the chewing gum containing PME for 10 minutes for one month after each meal within 30 minutes. Salivary secretion was collected by the participants between 8 and 10 a.m in the morning in the research office. For the measurement of secretory IgA and total protein concentrations in the saliva, indirect enzyme-linked immunosorbent assay(ELISA) was used. Results: The salivation stimulation rate was significantly increased after four weeks of masticating chewing gum containing PME after each meal(p<0.001). Mastication of chewing gum containing PME for four weeks decreased the concentration of secretory IgA much more significantly than that after mastication for one week(p=0.003). The concentration of total protein in the saliva was decreased after four weeks in the experimental and control groups. Conclusions: Mastication of chewing gum containing PME stimulated salivary secretion and led to oral disease prevention in patients with xerostomia. Furthermore, it seems to be urgent to seek measures that can be utilized in intervention for patients with xerostomia.

• 대한수의학회지
• /
• 제39권3호
• /
• pp.538-544
• /
• 1999

To elucidate the involvement of interleukin-$1{\beta}$ converting enzyme (ICE) in the course of experimental autoimmune encephalomyelitis (EAE), we induced EAE by immunizing rats with an emulsion of rat spinal cord homogenate with complete Freund's adjuvant supplemented with Mycobacterium tuberculosis (H37Ra, 5mg/ml) and then examined the expression of ICE in the spinal cord of rats with EAE. In normal rat spinal cords, ICE is constitutively, but weakly, expressed in ependymal cells, neurons, and some neuroglial cells. In EAE, many inflammatory cells are positive for ICE, and the majority of ICE+ cells were identified as ED1+ macrophages. During this stage of EAE, the number of ICE+ cells in brain cells, including neurons and astrocytes, increased and these cells also had increased ICE immunoreactivity. These findings suggest that the upregulation of ICE in both brain cells and invading hematogenous cells is stimulated by a secretory product from inflammatory cells, and that this enzyme is involved in the pathogenesis of EAE via the production of IL-1 beta.