• Journal of Ginseng Research
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• 제43권2호
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• pp.218-225
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• 2019

Background: Streptococcus pneumoniae, more than 90 serotypes of which exist, is recognized as an etiologic agent of pneumonia, meningitis, and sepsis associated with significant morbidity and mortality worldwide. Immunization with a pneumococcal pep27 mutant (${{\Delta}}pep27$) has been shown to confer comprehensive, long-term protection against even nontypeable strains. However, ${{\Delta}}pep27$ is effective as a vaccine only after at least three rounds of immunization. Therefore, treatments capable of enhancing the efficiency of ${{\Delta}}pep27$ immunization should be identified without delay. Panax ginseng Mayer has already been shown to have pharmacological and antioxidant effects. Here, the ability of Korean Red Ginseng (KRG) to enhance the efficacy of ${{\Delta}}pep27$ immunization was investigated. Methods: Mice were treated with KRG and immunized with ${{\Delta}}pep27$ before infection with the pathogenic S. pneumoniae strain D39. Total reactive oxygen species production was measured using lung homogenates, and inducible nitric oxide (NO) synthase and antiapoptotic protein expression was determined by immunoblotting. The phagocytic activity of peritoneal macrophages was also tested after KRG treatment. Results: Compared with the other treatments, KRG significantly increased survival rate after lethal challenge and resulted in faster bacterial clearance via increased phagocytosis. Moreover, KRG enhanced ${{\Delta}}pep27$ vaccine efficacy by inhibiting reactive oxygen species production, reducing extracellular signal-regulated kinase apoptosis signaling and inflammation. Conclusion: Taken together, our results suggest that KRG reduces the time required for immunization with the ${{\Delta}}pep27$ vaccine by enhancing its efficacy.

• BMB Reports
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• 제44권4호
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• pp.256-261
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• 2011

PEP-1 peptide has been used for transduction of native protein into mammalian cells. This work describes the findings that the fusion of PEP-1 to target proteins led to protein truncation likely in a non-protein-specific manner. Approximately 75% of PEP-1-MsrA fusion protein was truncated in the N-terminal region of MsrA between Lys-27 and Val-28 during expression in Escherichia coli and purification. This large protein truncation was also observed in another PEP-1 fused protein, PEP-1-MsrB2, in the N-terminal region of MsrB2. The full-length PEP-1-MsrA protein was rapidly transduced into keratinocyte cells within 15 min. The transduced PEP-1-MsrA was functionally active and could protect skin cells against oxidative stress- and ultraviolet radiation-induced cell death. Collectively, our data demonstrated the protective roles of MsrA in skin cells and, moreover, may raise a concern of protein truncation caused by fusion of PEP-1 about the general use of this peptide for protein transduction.

• Journal of Microbiology and Biotechnology
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• 제21권12호
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• pp.1345-1351
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• 2011

Vancomycin therapy failure due to the emergence of tolerance in pneumococci is increasing. The molecular mechanism of tolerance is not clear, but lytA and $pep^{27}$ are known to be involved. Our aim was to evaluate the expression of both genes in vancomycin-tolerant Streptococcus pneumoniae (VTSP) strains. Eleven VTSP strains from a total of 309 clinical isolates of S. pneumoniae from 1997 to 2006 were classified according to the criteria of Liu and Tomasz. All VTSP strains were evaluated for susceptibility according to CLSI criteria, serotype by the Quellung test, and clonality by PFGE. The expressions of lytA and $pep^{27}$ were analyzed in different growth phases by RT-PCR with and without vancomycin. Eighty-two percent of VTSP strains showed resistance to penicillin, and 100% were sensitive to vancomycin and cefotaxime. The most frequent serotypes of VTSP strains were 23F (4/11) and 6B (3/11). Clonal relationship was observed in only two strains. No significant changes were observed in $pep^{27}$ expression in the three phases of growth in VTSP strains with and without vancomycin. Interestingly, $pep^{27}$ expression in the stationary phase in the non-tolerant reference strain R6 was significantly higher. However, no significant differences in lytA expression were observed between VTSP and R6 strains during the phases of growth analyzed. The absence of changes in $pep^{27}$ expression in VTSP strains in the stationary phase may be related to their ability to tolerate high antibiotic concentrations, and thus, they survive and remain in the host under the antibiotic selective pressure reflected in therapeutic failure.

• Archives of Pharmacal Research
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• 제27권5호
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• pp.562-569
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• 2004

Self-assembling nanospheres of hydrophobized pullulan have been developed. Pullulan acetate (PA), as hydrophobized pullulan, was synthesized by acetylation. Carboxymethylated poly(ethylene-glycol) (CMPEG) was introduced into pullulan acetate (PA) through a coupling reaction using N, N'-dicyclohexyl carbodiimide (DCC). A synthesized PA-PEG-PA (abbreviated as PEP) conjugate was confirmed by Fourier transform-infrared (FT-IR) spectroscopy. Since PEP conjugates have amphiphilic characteristics in aqueous solution, polymeric nanoparticles of PEP conjugates were prepared using a simple dialysis method in water. From the analysis of fluorescence excitation spectra primarily, the critical association concentration (CAC) of this conjugate was found to be 0.0063 g/L. Observations by scanning electron microscopy (SEM) showed the spherical morphologies of the PEP nanoparticles. The particle size distribution of the PEP conjugates was determined using photon correlation spectroscopy (PCS) and the intensity-average particle size was 193.3 ${\pm}$ 13.53 nm with a unimodal distribution. Clonazepam (CNZ), as a model drug, was easy to entrap into polymeric nanoparticles of the PEP conjugates. The drug release behavior was mainly diffusion controlled from the core portion.

• Molecules and Cells
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• 제27권2호
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• pp.159-166
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• 2009

Myocardial ischemia-reperfusion injury is a medical problem occurring as damage to the myocardium following blood flow restoration after a critical period of coronary occlusion. Oxygen free radicals (OFR) are implicated in reperfusion injury after myocardial ischemia. The antioxidant enzyme, Cu, Zn-superoxide dismutase (Cu, Zn-SOD, also called SOD1) is one of the major means by which cells counteract the deleterious effects of OFR after ischemia. Recently, we reported that a PEP-1-SOD1 fusion protein was efficiently delivered into cultured cells and isolated rat hearts with ischemia-reperfusion injury. In the present study, we investigated the protective effects of the PEP-1-SOD1 fusion protein after ischemic insult. Immunofluorescecnce analysis revealed that the expressed and purified PEP-1-SOD1 fusion protein injected into rat tail veins was efficiently transduced into the myocardium with its native protein structure intact. When injected into Sprague-Dawley rat tail veins, the PEP-1-SOD1 fusion protein significantly attenuated myocardial ischemia-reperfusion damage; characterized by improving cardiac function of the left ventricle, decreasing infarct size, reducing the level of malondialdehyde (MDA), decreasing the release of creatine kinase (CK) and lactate dehydrogenase (LDH), and relieving cardiomyocyte apoptosis. These results suggest that the biologically active intact forms of PEP-1-SOD1 fusion protein will provide an efficient strategy for therapeutic delivery in various diseases related to SOD1 or to OFR.

• BMB Reports
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• 제42권3호
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• pp.136-141
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• 2009

Familial Amyotrophic lateral sclerosis (FALS) is a progressive neurodegenetative disorder induced by mutations of the SOD1 gene. Heat shock protein 27 (HSP27) is well-defined as a stress-inducible protein, however the its role in ALS protection has not yet been established. To investigate the role HSP27 may have in SOD1 mutant-mediated apoptosis, human SOD1 or HSP27 genes were fused with a PEP-1 peptide in a bacterial expression vector to produce a genetic in-frame fusion protein, which was then transduced into cells. We found the purified PEP-1-HSP27 fusion proteins can be transduced efficiently into neuronal cells and protect against cell death by enhancing mutant SOD1 activity. Moreover, transduced PEP-1-HSP27 efficiently prevents protein aggregation produced by oxidative stress. These results suggest that transduced HSP27 fusion protein may be explored as a potential therapeutic agent for FALS patients.

• Journal of Microbiology and Biotechnology
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• 제32권6호
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• pp.709-717
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• 2022

Allergic rhinitis (AR), one of the most common inflammatory diseases, is caused by immunoglobulin E (IgE)-mediated reactions against inhaled allergens. AR involves mucosal inflammation driven by type 2 helper T (Th2) cells. Previously, it was shown that the Streptococcus pneumoniae pep27 mutant (Δpep27) could prevent and treat allergic asthma by reducing Th2 responses. However, the underlying mechanism of Δpep27 immunization in AR remains undetermined. Here, we investigated the role of Δpep27 immunization in the development and progression of AR and elucidated potential mechanisms. In an ovalbumin (OVA)-induced AR mice model, Δpep27 alleviated allergic symptoms (frequency of sneezing and rubbing) and reduced TLR2 and TLR4 expression, Th2 cytokines, and eosinophil infiltration in the nasal mucosa. Mechanistically, Δpep27 reduced the activation of the NLRP3 inflammasome in the nasal mucosa by down-regulating the Toll-like receptor signaling pathway. In conclusion, Δpep27 seems to alleviate TLR signaling and NLRP3 inflammasome activation to subsequently prevent AR.

• Pediatric Infection and Vaccine
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• 제23권3호
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• pp.180-187
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• 2016

목적: 본 연구는 병원 내에서 의도치 않게 수두-대상포진 바이러스(varicella zoster virus [VZV]) 감염 환자에 노출된 의료진과 소아 입원 환자의 사례를 대상으로 노출 후 예방 조치와 그에 따른 2차 수두 감염 발생 여부를 분석하고자 하였다. 방법: 2010년 1월부터 2015년 12월까지 서울대학교 어린이병원에 입원한 수두 혹은 대상포진 환자중 초기에 적절한 격리 조치가 이루어지지 않았던 사례와 노출자를 대상으로 하였다. 노출자의 VZV에 대한 면역력과 면역 저하 상태의 유무에 따라 노출 후 예방 조치를 시행하였다. 의무기록을 통하여 사례 환자와 노출자들의 임상 정보 및 2차 감염 발생 여부를 조사하였다. 결과: 2010년부터 2015년까지 147명의 VZV 감염 환자가 입원하였고 이 중 의도치 않게 노출되었던 환자는 13명이었다. 이 중 5명(38.5%)의 사례 환자는 수두 백신 접종력이 확인되었다. 총 86명의 환자가 다인용 병실에서 사례 환자에 노출되었고, 62.8% (54/86)에서 VZV에 대한 면역력이 있었다. 27명의 노출 환자에게 노출 후 예방 조치를 시행하였으며, VZIG를 투약받은 환자는 23명이었고 수두 백신을 접종받은 환자는 4명이었다. 2차 수두가 발병한 환자는 4명으로, 예방 조치를 받지 않은 소아 1명과 예방 조치를 받은 27명 중 3명에서 감염이 확인되었다. 이들은 모두 한 명의 사례 환자에게 노출되었다. 2차 수두 감염률은 4.7% (4/85)이었고, 노출 후 예방 조치를 받은 환자 중 2차 감염률은 11.1% (3/27)이었다. 면역 기능이 정상인 환자에서 2차 수두 감염률은 1.9%, 면역 저하 환자에서는 9.7%이었다. 결론: 수두의 진단이 지연되면 병원에서 VZV에 노출되는 사례가 발생할 수 있으며 이로 인하여 감수성이 있는 소아나 면역 저하자에게 수두가 2차적으로 발생할 수 있다. VZV에 대한 면역력 여부를 기반으로 노출 후 예방 조치 여부를 결정하는 국내 기준의 적정성을 재고할 필요가 있다.

• 식물병연구
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• 제24권1호
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• pp.66-74
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• 2018

국내 노지 고추에서의 바이러스 발생률 및 발병현황을 조사하기 위해 2015-2016년까지 포장조사를 수행하였다. 시료 채집은 6월부터 9월까지 월별로 실시하였고 노지 포장에서 바이러스 병징을 보이는 고추에 대해 2015년 424개, 2016년 368개의 시료를 채집하였다. 감염여부는 7종 바이러스(오이모자이크바이러스, 잠두위조바이러스2, 토마토반점위조바이러스, 사탕무황화바이러스, 고추모틀바이러스, 감자바이러스 Y, 고추약한모틀바이러스)에 대한 유전자 진단법을 이용하여 검정하였다. 그 결과, 바이러스 발병률은 2015년에는 91.7%, 2016년에는 98%로 조사되었고, 조사지역에 따라 발생하는 바이러스의 종류는 차이가 있었으나 7종 바이러스가 모두 발생됨을 확인하였다. 2015년 바이러스 종류별 발생률은 CMV, BBWV, BWYV, PMMoV, TSWV, PepMoV 및 PVY가 각각 73.8%, 68.3%, 46.9%, 14.6%, 12.7%, 6.6% 및 3.3%이었으며 2016년 바이러스 발생률은 CMV, BBWV2, BWYV, TSWV, PMMoV, PepMoV 및 PVY가 73.3%, 71.4%, 34.7%, 27.9%, 19.2%, 13.5% 및 3.5%로 조사되었다. 바이러스 발생양상을 분석한 결과, 전체 채집시료의 복합감염 비율이 2015년에는 83%, 2016년에는 86.7%로 단독감염보다 대부분 복합감염의 형태로 발병됨을 알 수 있었다.

• Clinical Endoscopy
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• 제56권4호
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• pp.490-498
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• 2023

Background/Aims: Post-endoscopic retrograde cholangiopancreatography pancreatitis (PEP) is the most common and serious complication of endoscopic retrograde cholangiopancreatography. To prevent this event, a unique precutting method, termed opening window fistulotomy, was performed in patients with a large infundibulum as the primary procedure for biliary cannulation, whereby a suprapapillary laid-down H-shaped incision was made without touching the orifice. This study aimed to assess the safety and feasibility of this novel technique. Methods: One hundred and ten patients were prospectively enrolled in this study. Patients with a papillary roof size ≥10 mm underwent opening window fistulotomy for primary biliary access. In addition, the incidence of complications and success rate of biliary cannulation were evaluated. Results: The median size of the papillary roof was 6 mm (range, 3-20 mm). Opening window fistulotomy was performed in 30 patients (27.3%), none of whom displayed PEP. Duodenal perforation was recorded in one patient (3.3%), which was resolved by conservative treatment. The cannulation rate was high (96.7%, 29/30 patients). The median duration of biliary access was 8 minutes (range, 3-15 minutes). Conclusions: Opening window fistulotomy demonstrated its feasibility for primary biliary access by achieving great safety with no PEP complications and a high success rate for biliary cannulation.