Utilisation of ferroproteins by Candida albicans during candidastasis by apo-ovotransferrin

한용문 동덕여자대학교 종합약학연구소 2001 동덕약학연구지 Vol.5 No.-

암포테리신 B의 Candida albicans 성장 억제에 대한 베르베린의 상승효과

한용문 동덕여자대학교 종합약학연구소 2002 동덕약학연구지 Vol.6 No.-

In this study we tested a synergistic effect of berberine isolated from Coptidis Rhizoma against growth of Candida albicans. Berberine was extracted in 80% methanol. By agar diffusion susceptibility assay our berberine extract showed growth-inhibitory activity against Candida albicans. With regard to amount of this compound, the anticandidal effect was not significant as compared to anticandidal effect by amphotericin B (Am B). However, in case Am B was combined with berberine, anticandidal activity of the Am B was enhanced up to 4 times more than Am B alone. These data indicate that berberine was synergistic with Am B. Further, the data suggest the combination effect can lessen side effects due to toxicity of Am B at high doses.

Effects of Sugar Additives on Protein Stability of Recombinant Human Serum Albumin during Lyophilization and Storage

한용문,진병석,Sang-Bong Lee,Yeowon Sohn,Jee-Won Joung,이주희 대한약학회 2007 Archives of Pharmacal Research Vol.30 No.9

Few researches on the protein stabilization of recombinant human serum albumin (rHSA) have been done. In the present study, we assessed the impact of sugar lyoprotectants on the protein stability of lyophilized rHSA (65 KDa) in the solid state. For the assessment, rHSA was formulated with sucrose and trehalose, respectively, alone or in combination with mannitol, which were lyophilized and stored at 35oC. Degradation and aggregation of the resulting lyophilized formulations was analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Induction of amorphous state by the lyophilactants with rHSA was determined by differential scanning calorimetry (DSC). The protein secondary structure of the rHSA in the formulations was analyzed by Fourier transform infrared spectroscopy (FT-IR). Results from SDS-PAGE analysis displayed that mannitol formulation caused aggregation resulting in a few bands that were greater than 65 KDa, whereas sucrose and trehalose formulations revealed no such aggregation. However, the aggregation of the protein decreased when mannitol was combined with sucrose or trehalose. DSC measurement supported the electrophoresis data showing that sucrose and trehalose formed complete amorphous state, but mannitol induced a partial amorphous state. These data indicate during lyophilization the most effective protein protection against aggregation was provided by sucrose and trehalose. The protection lasted during 4 months storage at 35oC. FT-IR analysis displayed that the sucrose formulation inhibited deamidation. In conclusion, our data suggest that sucrose and trehalose as additives seems to be sufficient to protect from lyophilization of rHSA protein and also maintain its stability in the solid state during storage.

18β-Glycyrrhetinic Acid의 면역보조제효능에 의한 항 전신성캔디다증 효과

한용문 대한약학회 2008 약학회지 Vol.52 No.6

The role of antibody in the fungal infections is controversial. However, our previous reports showed a certain epitope in Candida albicans cell wall (CACW) induces protective antibody. A major problem is that the epitope isolation requires tremendous time with high cost. This aspect led us to investigate a simple way inducing protective antibodies against C. albicans. In the present study, we determined if 18β-glycyrrhetinic acid (18β-GA) from Glabrae Radix (a family of Leguminosae) has immunoadjuvant activity. Data displayed that the 18β-GA suppressed proliferations of both T- and Blymphocytes at high concentrations, whereas below 20 μM concentration the compound supported the proliferations. These observations indicate that 18β-GA has immunoregulatory activity. Based on this observation, an immunoadjuvant effect was examined at the low concentration. Results from animal experiments showed that CACW combined with or without 18β- GA produced the anti-C. albicans antiserum in mice. Nevertheless, the CACW combined with 18β-GA formula only protected mice against disseminated candidiasis (P<0.05). These data implicate that 18β-GA has immunoadjuvant activity, which may provoke the CACW antigen to induce protective antibody. Currently, we are investigating possible mechanism of how the 18β-GA provokes such protective immunity against the disseminated disease.

캔디다질염에 효과가 있는 단항체에 대한 호중구의 역할

한용문 대한약학회 2003 약학회지 Vol.47 No.3

As previously reported, an immunoglobulin M (IgM) monoclonal antibody (MAb) B6.1, specific for a cell wall B-l,2-mannotriose, was protective against vaginal infection due to Candida albicans when mice were treated with the antibody. In this study, the role of neutrophil was examined in the protective effect of MAb B6.1 against vaginal infection. To deplete neutrophils, mice were given intravenously rat anti-mouse neutrophile MAb RB6-8C5 prior to intraperitoneal administration of MAb B6.1 to these mice. The mice were examined for antibody in their reproductive tract. By an ELISA, MAb B6.1 was found in the vaginal homogenates, but no antibody was detected in vaginal lavage materials. The neutropenia was induced by a single dose of the anti-neutrophil antibody, but lymphocytes were also partially depleted. The protective effect of MAb B6.1 was decreased when mice pretreated with MAb RB6-8C5 were given the anti-Candida antibody before challenge with C. albicans yeast cells intravaginally. These results show that neutrophils are involved in the MAb B6.1 protection against Candida vaginal infection.

Chlorogenic Acid와 Fluconazole 병용에 의한 항진균작용의 상승효과

한용문,유정임,Han, Yongmoon,Rhew, Zheong-Imm 대한약학회 2016 약학회지 Vol.60 No.4

In this present study, we determined the ability of chlorogenic acid (CRA), a polyphenolic compound, to potentiate the antifungal activity of fluconazole (FCZ) against Candida albicans. The determination was investigated by a microdilution susceptibility method and in a murine model of cutaneous candidiasis due to C. albicans. Results showed that CRA had a dose-dependent antifungal activity; and when combined with FCZ, the antifungal activity was synergistic as determined by the microdilition susceptibility method. For example, a combination of CRA at $25{\mu}g/ml$ plus FCZ at $0.1{\mu}g/ml$ was approximately 3 times more effective than antifungal activity of FCZ alone at the same dose (P<0.01). This potentiation by CRA was almost equivalent to antifungal activity by FCZ alone at $0.5{\mu}g/ml$, displaying that the CRA potentiates FCZ's antifungal activity by almost 5 times (P<0.01). Moreover, this combination demonstrated synergic activity against the cutaneous disease as well, resulting in approximately 3 times more greater antifungal activity than FCZ alone at $0.1{\mu}g/mouse$ (P<0.01). This suggests that the combination therapy can reduce side effects caused by FCZ in clinical situations. In summary, CRA has a synergistic antifungal activity, which can be produced by combining CRA with FCZ. Therefore, our data strongly indicate that CRA can be a potential candidate as an antifungal agent for combination therapy.

페렴구균 전신감염에 대한 협막. 표면단백질 접합백신의 효과

한용문,이주희,Han , Yong-Moon,Lee , Jue-Hee 대한약학회 2004 약학회지 Vol.48 No.6

ln the present work to determine effect of a Streptococcus pneumoniae conjugate vaccine, S.pneumoniae capsule attached to the surface protein (JY-Pol) was ex amined. This JY-Pol contained approximately 92% and 6% carbohydrate and protein, respectively. Gel electrophoresis revealed the presence of the surface protein in the JY-Pol. By the double immunodiffusion and isotyping ELISA analyses, administration of JY-Pol that was adsorbed to alum adjuvant (JY-Pol/Alum) into mice induced IgM, IgG, and IgA specific for the S.pneumoniae capsule. The ATCC capsular polysaccharide adsorbed to alum (ATCC-Pol/Alum) provoked only IgM in mice. In survival tests, mice that were immunized with the JY-Pol/Alum before intravenous challenge with live S.pneumoniae survived entire period of 46 day-observation, whereas all mice that received ATCC-Pol/Alum or only diluent instead of the vaccination died within 5 and 12 days, respectively. Results from footpad-edema test showed that JY-Pol/Alum formula provoked the cellular immunity as determined by swelling of the mouse footpad. These data indicate that the naturally conjugated JY- Pol enhances resistance of mice against disseminated pneumococcal disease due to S.pneumoniae by both humoral and cellular immune responses.

$18{\beta}$-Glycyrrhetinic Acid의 항 감염성관절염효과

한용문,Han, Yong-Moon 대한약학회 2007 약학회지 Vol.51 No.6

A polymorphic fungus, Candida albicans, causes various forms of infections such as disseminated candidiasis and vaginitis. Recent reports indicate that the fungus is a main etiological agent for the arthritis. In search of new sources for treatment of the fungal arthritis, we examined $18{\beta}$-glycyrrhetinic acid ($18{\beta}$-GA) against C. albicans-caused septic arthritis. The compound is isolated from Glycyrrhizae Radix that is known to have various immunomodulating activities and is one of the most popular herbal medicines. For induction of animal model of a septic arthritis, mice were given an emulsion form of C. albicans cell wall mixed with Complete Freund's Adjuvant (CFA) via footpad-injection. To determine prophylactic and therapeutic effects, the component was given to the animals before or after the induction of the arthritis, respectively. Data showed that intraperitoneal administration of $18{\beta}$-GA resulted in reduction of the inflammation, indicating the component had both prophylactic and therapeutic activities. For investigation of mechanism of the $18{\beta}$-GA, inhibitory effects on NO (nitiric oxide) and on T-lymphocyte proliferation were determined. Results demonstrated that $18{\beta}$-GA suppressed NO production from LPS (lipopolysaccharide)-treated macrophages and also inhibited proliferation of Con A (concanavalin A)activated T-cells. Taken together, $18{\beta}$-GA, a pentacyclic triterpene, has anti-arthritic activity against C. albicans-caused septic arthritis, possibly by blocking NO production and T-cell suppression.

Apotransferrin has a second mechanism for anticandidal activity through binding of Candida albicans

한용문 대한약학회 2014 Archives of Pharmacal Research Vol.37 No.2

It has been reported that transferrin has antibacterialand antifungal activities via iron chelation in theenvironment surrounding the microbes. In the presentstudy, we investigated whether the binding of transferrin toCandida albicans mediates growth inhibition. By usingcultures that contained iron-free (apo)transferrin glycoproteineither in contact with candidal cells or separatedfrom candidal cells by a dialysis membrane, we distinguishedthe growth inhibition by transferrin–cell interactionfrom that of simple iron chelation. Maximal growthinhibition always occurred when the apotransferrin interacteddirectly with the cells. Additionally, there was partialinhibition even when candidal cells were in contact withiron-saturated transferrin. Binding studies with 59Fe3?radiolabeled-transferrin indicated that the apo-protein canbind to the candidal cell surface. The binding sites weresaturable and it was dose dependent. Chemicals (hydrogenperoxide, dithiothreitol, sodium dodecyl sulfate) blockedtransferrin binding to C. albicans, and among the three,hydrogen peroxide (HP) was the most effective for theblocking. When HP-treated yeast cells were added to theculture that was pretreated with apotransferrin, candidalcell growth increased by 5-fold as compared to the growthof HP-untreated candidal cells under apotransferrin-regulation(P\0.05). Combined all data together, it wasconcluded that transferrin has a second mechanism of anticandidalactivity that is mediated by binding to the surfaceof C. albicans yeast cells.

Utilization of ferroproteins by Candida albicans during candidastasis by apo-ovotransferrin

한용문 동덕여자대학교 종합약학연구소 2001 동덕약학연구지 Vol.5 No.-

Recent reports have shown that some pathogenic bacteria can obtain iron from ferroproteins, such as cytochrome C, ferritin, hemin, hemoglobin and myoglobin. These studies prompted to determine if the opportunistic pathogenic fungi, Candida albicans, can utilize ferroproteins to circumvent the iron-regulatory effect of ovotransferrin. Assays to measure growth stimulation in vitro by the ferroproteins were conducted as follows: C. albicans was cultured in iron-free (pretreated with apo-ovotransferrin for 24 h) culture medium. Once C. albicans yeast cell growth reached stasis from iron starvation, individual ferroproteins were added to the culture media. The results suggested that hemin, hemoglobin, and myoglobin were able to support a partial growth recovery. Additional studies with haptoglobin, a serum protein that interacts with the globin moiety of certain ferroproteins, established that C. albicans was able to obtain iron from the haptoglobin-ferroprotein complexes. This result suggests that the heme group may be the iron source from the ferroproteins.