Streptococcus mutans has been implicated as primary causative agents of dental caries by insoluble glucan (IG) in human and experimental animals. An attempt was made to search for the ${\alpha}$-1,3 glucanase that degrades IG produced by S. mutans. ${\alpha}$-1,3 glucanase was detected in the culture supernatant of microorganisms, which are isolated from soils on agar medium containing IG as a sole carbon source. This Streptomyces sp. hydrolysed IG produced by immobilized S. mutans and was named as Y9373. This enzyme required ${\alpha}$-1,3 glucan (IG) as an inducer. The optimum conditions for enzyme production were studied. The enzyme was purified by 30~70% $(NH_4)_2SO_4$ precipitation, anion exchange chroma tography on DEAE-cellulose and gel filtration on Sepadex G-75. The purified enzyme has a specific activity of 7840.0 U/mg protein giving 32.1-fold purification and final yield of 0.53%. The molecular weight was estimated to be about 22.5 kDa by SDS-PAGE. The optimum pH and temperature for enzyme reaction were 6.5 and 37$^{\circ}C$, respectively and the enzyme was relatively stable at the temperature below 60$^{\circ}C$. The activity of purified enzyme was enhanced by adding $Co^{2+},\;Mn^{2+}\;and\;Mg^{2+}$ into the medium, whereas inhibited by adding $Hg^{2+},\;Zn^{2+}$ and SDS. The $K_m\;and\;V_{max}$ value of ${\alpha}$-1,3 glucanase for IG were estimated to be 2.50 mM and 0.0431 mM/min, respectively. The thin layer chromatographic analysis of hydrolysates from IG with ${\alpha}$-1,3 glucanase showed that glucose was the main product of reaction. This enzyme activity was about 14 times higher than marketing dextranase as preventive agent against artificial dental caries by S. mutans in TH medium including 5% sucrose after 30 minutes.
Toxoplasma gondii has been shown to result in life-threatening encephalitis in immunocompromised patients after reactivation of dormant parasites. In order to obtain information on immune responses related to this phenomenon, BALB/c mice were infected with 25 cysts of the 76K strain of T. gondii, then, treated orally with dexamethasone (Toxo/Dexa-treated group) in order to reactivate the chronic toxoplasmosis. None of the T. gondii-infected mice died during the experimental periods, whereas the Toxo/Dexa-treated mice evidenced a significant attenuation of survival periods. Toxoplasma-specific IgG2a, IgA and IgM titers in sera were significantly depressed in the Toxo/Dexa-treated mice; however, the IgG1 sera titers were similar to those seen in the Toxoplasma-infected mice. The percentages of CD4+ and $CD8\alpha+$ T cells in the Toxo/Dexa-treated mice were significantly reduced 2 weeks after dexamethasone treatment. $IFN-\gamma$ and IL-10 production levels in the Toxo/Dexa-treated mice were depressed significantly, whereas IL-4 production was increased temporarily. The expression levels of the Toxoplasma-specific P30 and B1 genes were found to have been increased in the Toxo/Dexa-treated mice in comparison with the Toxoplasma-infected mice. Collectively, the findings of this study demonstrate that reactivation of murine toxoplasmosis as the result of dexamethasone treatment induced a depression in Th1 immune responses, whereas Th2 immune responses were not significantly influenced.
Immunogenicity of dendritic cell-derived exosomes stimulated with Toxoplasma gondii lysates (TLA exo), mixed with cholera toxin as an adjuvant, was investigated in mice immunized via 2 mucosal routes (ocular vs intranasal). BALB/c mice were injected 3 times with TLA exo vaccine at 2 week interval, and the levels of IgG in serum and IgA in tear, saliva, feces, and vaginal wash were measured. To observe the expression of T. gondii-specific B1 gene, mice infected with ME49 T. gondii cysts were immunized with TLA exo or PBS exo (not stimulated with TLA), and their brain tissues were examined. The mice vaccinated via intranasal route elicited significantly higher humoral and mucosal immune responses compared with mice treated with PBS alone. Also, mice immunized via ocular route (by eyedrop) induced significantly higher T. gondii-specific IgG in serum and IgA in tear and feces in comparison with PBS controls. B1 gene expression was significantly lower in TLA exo vaccinated mice than in PBS or PBS exo vaccinated mice. These results demonstrated that ocular immunization of mice with TLA exo vaccine has the potential to stimulate systemic or local antibody responses. This study also highlighted an advantage of an eyedrop vaccine as an alternative for T. gondii intranasal vaccines.
This study was conducted to assess the nutritional status of the elderly from low income areas and its relationship with the indices of immunity. Ninety nine(male : 28, female : 71) healthy, subjects aged over 65 years staying in homes in Seoul were assessed with regard to their dietary intake and biochemical measurements. Their mean daily energy intakes were 1256kcal and 109kcal for males and females respectively. Mean serum IgG were 2356mg/dl in men and 2161mg/dl for women, higher than the normal range. Mean serum $C_3$ were 120.9mg/dl and 130.6mg/dl for males and females respectively and mean $C_4$ were 30.6mg/dl and 31.9mg/dl. The mean $C_3$ and $C_4$ were within the normal range and the proportion of subjects whose $C_3$ and $C_4$ were at a higher level than the upper limit were 15.4$\%$ and 16.3$\%$ respectively. Complement $C_3$ showed significant correlation with serum albumin(p〈0.01), serum cholesterol(p〈0.00l, serum TG(p〈0.05), LDL-C(p$\ll$$\ll$$\ll$$\ll$$\ll$$\ll$$\ll$$\ll$$\ll$$\ll$$\ll$$\ll$$\ll$q0.001), VLDL-C(p$\ll$0.05), serum copper(p$\ll$0.05) and serum Ca(p$\ll$0.01). (Korean J Community Nutrition 2(4) : 547-555, 1997)
In order to obtain more specific antigenic preparation for the diagnosis of human paragonlmiasis, crude saline extract of whole worm (=PwWWE), secretory.excretory components (PwSEC) and secretion-excretion-free somatic extract (PwSM) of 12 week-old ParagoninBus westermani were filtrated through Sephadex G-200 gel column. The adult Paragonimus worms were obtained from expevimentally infected doge. A total of 11 antigenic solutions was Iyophilised or diluted to adjust protein content of 1mg/ml. To evaluate the antigenicity of crude antigens and fractions, micro-ELISA was done with the sera from P westermani in(ected cases, C. sinensis infected cases and non-infected control cases to detect Paragonimus specific IgG antibody. The results were as follows: 1. When the PwWWE was filtrated through Sephadex G-200 gel, it was separated into three fractions; PwWWE Fr. 1, PwWWE Fr. 2 and PwWWE Fr. 3. The percentage of protein content was 28.0%, 21.6% and 50.4% respectively. The PwSM was also. separated into three fractions; PwSM Fr. 1, PwSM Fr. 2, PwSM Fr. 3. and their percentage of protein content was 41.3%, 38.6% and 20.1%. However, the PwSEC showed different fractionation pattern; i.e. fraction 1 (=PwSEC Fr.1) and 3 (PwSEC Fr. 3) without fraction 2. The percentage of protein content was 14.0% in PwSEC Fr. 1 and 86.0% in PwSEC Fr. 3. 2. When the antigenicity of each Paragonimus crude antigen and fractionated antigen was evaluated for specific IgG aritibody by micro-ELISA in 10 human paragonimiasis sera, PwSEC Fr, 1 was the most potent antigen showing the mean absorbance 1.98. The PwWWE Fr. 1, PwSEC, PwWWE were next to that: their mean absorbance were 1.72, 1.38 and 0.83 respectively. The antigenicity of fractions 2 and 3 was much weaker in binding specific IgG antibody. 3. When the antigens were reacted in micro-ELISA with 10 human clonorchiasis sera, most antigens showed weak reactivity. Each fraction 1 of crude antigens reacted higher than other fractions or crude antigens; the mean absorbance was 0.17 in fraction 1, but in others the absorbances were about 0.06. 4. With non-infected control sera, the result of micro-RLISA revealed almost same pattern with those of the clonorchiasis sera. From the above results, it became apparent that PwWWE Fr. 1, especially PwSEC Fr. 1 was the most potent antigen reacted with Paragonisfaus specific IgG antibody.
The present study is intended to observe the chronologic changes of experimental sparganosis by histopathological observation and detection of circulating anti-sparganum IgG antibody using ELISA. Each of 25 mice was infected with aye spargana, and they were examined after 1, 2, 4, 10 weeks or 6 months from infection. The followings are summarized results. - 1. The plerocercoids were detected in the subcutaneous tissue of the trunk, neck or axilla, but a few often extended into the skeletal muscle. The recovery rates were 72% at the first week, 80% at the second week, 95% at the fourth week, 92% at the tenth week and 100% at .the sixth month. The larvae grew slowly in both length and weight until 6 months. 2. Histopathologically, most of the larvae were observed alive in the soft tissue or skeletal muscle. Numerous eosinophils, neutrophils, Iymphocytes and plasma cells were infiltrated focally around the worms by the second week, but they surrounded the worms to form a layer of inflammatory reaction after 4 weeks of infection. Also histiocytes and fibroblasts began to appear around the inflammatory cells at 4 weeks. After 10 weeks, the worms encircled by a thin fibrous layer were found. After 6 months, the worms were surrounded by either fibrous tissue or active inflammatory cells. The inflammation looked more severe in the tracks left by the worms, rather than around the worms. 3, The level of anti-sparganum IgG antibody in the serum showed an increase by the fourth week, and a rapid and continuous increase was observed thereafter by the tenth week after infection. The high level of the IgG antibody was maintained up to 6 months forming a plateau curve. The present results suggest that the tissue reaction and antibody production in subcutaneous sparganosis become distinctive by the fourth week after infection.
The Transactions of the Korean Institute of Electrical Engineers D
/
v.55
no.2
/
pp.45-51
/
2006
In this paper, we propose a new architecture of Information Granulation based genetically optimized Self-Organizing Fuzzy Polynomial Neural Networks (IG_gSOFPNN) that is based on a genetically optimized multilayer perceptron with fuzzy polynomial neurons (FPNs) and discuss its comprehensive design methodology involving mechanisms of genetic optimization, especially information granulation and genetic algorithms. The proposed IG_gSOFPNN gives rise to a structurally optimized structure and comes with a substantial level of flexibility in comparison to the one we encounter in conventional SOFPNNs. The design procedure applied in the construction of each layer of a SOFPNN deals with its structural optimization involving the selection of preferred nodes (or FPNs) with specific local characteristics (such as the number of input variables, the order of the polynomial of the consequent part of fuzzy rules, and a collection of the specific subset of input variables) and addresses specific aspects of parametric optimization. In addition, the fuzzy rules used in the networks exploit the notion of information granules defined over system's variables and formed through the process of information granulation. That is, we determine the initial location (apexes) of membership functions and initial values of polynomial function being used in the premised and consequence part of the fuzzy rules respectively. This granulation is realized with the aid of the hard c-menas clustering method (HCM). To evaluate the performance of the IG_gSOFPNN, the model is experimented with using two time series data(gas furnace process and NOx process data).
Background: Korean ginseng is a well-known medicinal herb that has been widely used in traditional medicine to treat various diseases, including asthma. Ginseng can be classified as white ginseng (WG) or red ginseng (RG), according to processing conditions. In this study, the authors compared the efficacies of these two ginseng types in a mouse model of acute asthma. Methods: To produce the acute asthma model, BALB/c mice were sensitized with ovalbumin (OVA) and aluminum hydroxide, and then challenged with OVA. WG and RG extracts were administered to mice orally. The influences of WG and RG on airway hyperresponsiveness (AHR), immune cell distributions in bronchoalveolar lavage fluid (BALF), and OVA-specific immunoglobulin E (IgE), IgG1, and IgG2a in serum were investigated. Cytokine production by lymphocytes isolated from peribronchial lymph nodes and histopathological changes was also examined. Results: In OVA-sensitized mice, both WG and RG reduced AHR and suppressed immune cell infiltration in bronchoalveolar regions. BALF OVA-specific IgE levels were significantly lower in RG-treated OVAsensitized mice than in the OVA-sensitized control group. WG and RG also suppressed inflammatory cytokine production by peribronchial lymphocytes. Histopathological findings showed reduced inflammatory cell infiltration and airway remodeling (e.g., epithelial hyperplasia) in WG- and RG-treated OVA mice compared with OVA controls. Conclusion: In this study, WG and RG showed antiasthmatic effects in an OVA-sensitized mouse model, and the efficacies of RG were found to be better than those of WG.
Pollen has been used for Prevention or treatment of certain diseases such as diabetes, arthritis, or cancer in traditional medicine. In addition, pollen is under investigation as a host cell for a gene expression. This study was undertaken to evaluate the immunologic safety of pollen intake. BALB/c mice were administered with 500, 50,5, or 0.5 mg/kg bw of lily pollen for five times a week for four weeks through gastric intubation. Comparing the control mice administered with distilled water, no significant changes were observed in body weight gain, weight of liver, spleen, lung, and his-topathological findings of liver and kidney of the mice groups administered with the pollen. Plasma level of IgG1, IgG2a, and IgE was not different among the groups. When splenic B lymphocytes were stimulated in vitro with lipopolysaccharides for 7 days, level of IgGl and IgGwa produced in the culture supernatants was not significantly different among the groups. Furthermore, no significant alteration was observed in IL-4 and $IFN{\gamma}$ producing ability with splenic T lymphocytes stimulated in vitro with phytohemagglutinins for 48 hours between the pollen-administered and the control mice. Overall, this study suggests that the lily pollen intake is Inducing no significant modulation of humoral and cell-mediated immunity in mice.
Purpose: Hepatitis B virus (HBV) infection is among etiologies of secondary membranous nephropathy (MN) in pediatric patients. We evaluated expression of phospholipase A2 receptor (PLA2R), a specific target antigen of primary MN, in pediatric HBV-related MN. Methods: We retrospectively reviewed patients with biopsy-proven HBV-related MN from the renal biopsy registry and electronic medical records of Severance Hospital, Seoul, Korea, from 1993 to 2004. Paraffin-embedded human kidney tissues were retrieved and immunohistochemically stained for PLA2R. Results: Ten pediatric patients with 13 biopsied specimens were reviewed. The predominant pathological stage was stage II-III, and second was stage II. The intensity of staining for IgG was greatest, with less intense staining for IgM, IgA, C3, C4, and C1q. All the patients had angiotensin-converting enzyme inhibitor combined with glucocorticoid, and four patients converted to cyclosporine treatment from glucocorticoid monotherapy. Urinalysis of all the patients normalized after variable period. PLA2R staining was demonstrated in the outer glomerulus in 3 out of 13 biopsies, 2 of which were obtained from the same patient over a 5-year interval. Conclusions: PLA2R was expressed in a small number of cases diagnosed as pediatric HBV-related MN, indicating that some HBV-related MN cases may be primary MN concurrent with HBV infection.
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