• 대한수의학회지
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• 제63권1호
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• pp.6.1-6.9
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• 2023

Stroke is a major cause of death and long-term disability. Chlorogenic acid is a phenolic compound with a potent neuroprotective effect. γ-enolase is a phosphopyruvate hydratase found in mature neurons and plays an important role in neuronal survival. This study investigated whether chlorogenic acid regulates the expression of γ-enolase during cerebral ischemia. Middle cerebral artery occlusion (MCAO) was performed to induce cerebral ischemia. Adult male rats were used and chlorogenic acid (30 mg/kg) or phosphate buffered saline (PBS) was injected intraperitoneally 2 hours after MCAO surgery. Cerebral cortical tissues were collected 24 hours after MCAO surgery. Our proteomic approach identified the reduction of γ-enolase caused by MCAO damage and the mitigation of this reduction by chlorogenic acid treatment. Results of reverse transcription-polymerase chain reaction and Western blot analyses showed a decrease in γ-enolase expression in the PBS-treated MCAO group. However, chlorogenic acid treatment attenuated this decrease. Results of immunofluorescence staining showed the change of γ-enolase by chlorogenic acid treatment. These results demonstrated that chlorogenic acid regulates the γ-enolase expression during MCAO-induced ischemia. Therefore, we suggest that chlorogenic acid mediates the neuroprotective function by regulating the γ-enolase expression in cerebral ischemia and may be used as a therapeutic agent for brain diseases including stroke.

• 한국식품과학회지
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• 제54권1호
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• pp.1-7
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• 2022

본 연구에서는 형광등 빛 조사 하에 각종 감광제와 아미노산의 조합에 의한 NBT의 환원 특성을 조사하였다. 기존 SOD 활성 측정에 이용된 Rb-Met 반응계는 증류수나 Tris보다는 PBS 용매 상에서 가장 우수한 효과를 나타냈다. 빛 조사 하에 Rb에 의한 NBT 환원을 위해서는 Met과 His 등의 아미노산이 필요하며, 감광제로서는 isoalloxazine 계열의 Rb와 thiazine 계열의 MB가 효과적인 환원 반응을 유도했다. 하지만 각종 감광제나 아미노산의 조합, 그리고 반응 용매에 따라 NBT 환원정도가 상이하며, 특히 Rb-Met in PBS와 MB-His in Tris 반응계가 가장 큰 반응을 유도하였다. Rb에 의해 유도된 NBT 환원반응은 SOD 및 gallic acid에 의해 효과적으로 저해되었으나, Tris 상에서 MB-His에 의한 NBT 환원은 SOD 및 gallic acid에 의한 저해효과가 미미하여 Rb와는 다른 기작에 의해 NBT 환원이 유도되는 것으로 보인다. 본 결과는 감광제와 아미노산에 따라 빛 조사 하에 다양한 상호작용이 발생하며, NBT 환원을 유도하는데 superoxide anion 뿐만 아니라 다른 요인도 관여할 수 있음을 시사한다.

• Journal of Veterinary Science
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• 제23권1호
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• pp.7.1-7.14
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• 2022

Background: Enterotoxigenic Escherichia coli (ETEC) infection is a primary cause of livestock diarrhea. Therefore, effective vaccines are needed to reduce the incidence of ETEC infection. Objectives: Our study aimed to develop a multivalent ETEC vaccine targeting major virulence factors of ETEC, including enterotoxins and fimbriae. Methods: SLS (STa-LTB-STb) recombinant enterotoxin and fimbriae proteins (F4, F5, F6, F18, and F41) were prepared to develop a multivalent vaccine. A total of 65 mice were immunized subcutaneously by vaccines and phosphate-buffered saline (PBS). The levels of specific immunoglobulin G (IgG) and pro-inflammatory cytokines were determined at 0, 7, 14 and 21 days post-vaccination (dpv). A challenge test with a lethal dose of ETEC was performed, and the survival rate of the mice in each group was recorded. Feces and intestine washes were collected to measure the concentrations of secretory immunoglobulin A (sIgA). Results: Anti-SLS and anti-fimbriae-specific IgG in serums of antigen-vaccinated mice were significantly higher than those of the control group. Immunization with the SLS enterotoxin and multivalent vaccine increased interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) concentrations. Compared to diarrheal symptoms and 100% death of mice in the control group, mice inoculated with the multivalent vaccine showed an 80% survival rate without any symptom of diarrhea, while SLS and fimbriae vaccinated groups showed 60 and 70% survival rates, respectively. Conclusions: Both SLS and fimbriae proteins can serve as vaccine antigens, and the combination of these two antigens can elicit stronger immune responses. The results suggest that the multivalent vaccine can be successfully used for preventing ETEC in important livestock.

• 한국축산식품학회지
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• 제43권4호
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• pp.703-711
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• 2023

As an initial study to elucidate the molecular mechanism of how probiotics modulate macrophage activity, we monitored mRNA expression patterns in peritoneal macrophages (PMs) treated with two different strains of probiotics. After treatment with either Weissella cibaria WIKIM28 or Latilactobacillus sakei WIKIM50, total RNAs from PMs were isolated and subjected into gene chip analyses. As controls, mRNAs from vehicle (phosphate-buffered saline, PBS)-treated PMs were also subjected to gene chip analysis. Compared to vehicle (PBS)-treated PMs, WIKIM28-treated and WIKIM50-treated PMs exhibited a total of 889 and 432 differentially expressed genes with expression differences of at least 4 folds, respectively. Compared to WIKIM28-treated PMs, WIKIM50-treated PMs showed 25 up-regulated genes and 21 down-regulated genes with expression differences of more than 2 folds. Interestingly, mRNA transcripts of M2 macrophage polarization marker such as anxa1, mafb, and sepp1 were increased in WIKIM50-treated PMs comparing to those in WIKIM28-treated PMs. Reversely, mRNA transcripts of M1 macrophage polarization marker such as hdac9, ptgs2, and socs3 were decreased in WIKIM50-treated PMs comparing to those in WIKIM28-treated PMs. In agreement with these observations, mRNA expression levels of tumor necrosis factor-α and interleukin-1α were significantly reduced in WIKIM50-treated macrophages compared to those in WIKIM28-treated macrophages. These results may indicate that probiotics can be classified as two different types depending on their ability to convert macrophages into M1 or M2 polarization.

• Journal of Veterinary Science
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• 제24권4호
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• pp.53.1-53.12
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• 2023

Background: Mammalian orthoreovirus type 3 (MRV3), which is responsible for gastroenteritis in many mammalian species including pigs, has been isolated from piglets with severe diarrhea. However, the use of pig-derived cells as an infection model for swine-MRV3 has rarely been studied. Objectives: This study aims to establish porcine intestinal organoids (PIOs) and examine their susceptibility as an in vitro model for intestinal MRV3 infection. Methods: PIOs were isolated and established from the jejunum of a miniature pig. Established PIOs were characterized using polymerase chain reaction (PCR) and immunofluorescence assays (IFAs) to confirm the expression of small intestine-specific genes and proteins, such as Lgr5, LYZI, Mucin-2, ChgA, and Villin. The monolayered PIOs and three-dimensional (3D) PIOs, obtained through their distribution to expose the apical surface, were infected with MRV3 for 2 h, washed with Dulbecco's phosphate-buffered saline, and observed. Viral infection was confirmed using PCR and IFA. We performed quantitative real-time reverse transcription-PCR to assess changes in viral copy numbers and gene expressions linked to intestinal epithelial genes and antiviral activity. Results: The established PIOs have molecular characteristics of intestinal organoids. Infected PIOs showed delayed proliferation with disruption of structures. In addition, infection with MRV3 altered the gene expression linked to intestinal epithelial cells and antiviral activity, and these effects were observed in both 2D and 3D models. Furthermore, viral copy numbers in the supernatant of both models increased in a time-dependent manner. Conclusions: We suggest that PIOs can be an in vitro model to study the infection mechanism of MRV3 in detail, facilitating pharmaceutical development.

• Journal of Veterinary Science
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• 제24권6호
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• pp.74.1-74.15
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• 2023

Background: Antibiotic beads are used to treat local bacterial infections by delivering high drug concentrations to infected tissue. Objectives: This study examined the elution characteristics of metronidazole from metronidazole-calcium sulfate (MCa) and metronidazole-calcium-potassium sulfate (MCaK) beads over 20 days and the antibacterial efficacy of the beads after storage. Methods: The MCa and MCaK beads were prepared by mixing 250 mg of metronidazole and 10 g of calcium sulfate hemihydrate with water and a 3% potassium sulfate solution, respectively. The beads were placed in phosphate-buffered saline for the elution study. The metronidazole eluents were determined using high-performance liquid chromatography. The microstructures were examined by scanning electron microscopy (SEM), and the antimicrobial activity was evaluated by a microbioassay. Results: For the 20-day study, the total amount of metronidazole released was greater in the MCa beads than in the MCaK beads by 6.61 ± 0.48 mg (89.11% ± 3.04%) and 4.65 ± 0.36 mg (73.11% ± 4.38%), respectively. The amounts of eluted drugs from the MCa and MCaK beads were higher than the minimum inhibitory concentration at 0.5 ㎍/mL against anaerobic bacteria at both 20 days and 14 days. SEM showed that calcium crystals on the outer surface had dissolved after elution, and thinner calcium crystals were prominent in the MCaK beads. The MCa and MCaK beads exhibited antibacterial activity after setting, followed by storage at room temperature or 4℃ for 21 days. Conclusions: The MCa beads could release more drug than the MCaK beads, but all eluted metronidazole amounts were effective in controlling bacterial infections. Both metronidazole beads could be stored at ambient temperature or in a refrigerator.

• Journal of Dairy Science and Biotechnology
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• 제41권3호
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• pp.103-112
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• 2023

This study aimed to assess the effectiveness of 10 different pre-enrichment methods using Real-Time polymerase chain reaction (PCR) in support of the FDA method. When the initial Cronobacter spp. (Enterobacter sakazakii) inoculation was 7.2 CFU/g, the Ct values were observed in the following order: 21.37 (Enterobacteriaceae enrichment [EE] broth), 21.95 (brain heart infusion [BHI]), 22.72 (tryptic soy broth [TSB]), 23.02 (violet red bile lactose [VRBL]), 22.31 (TSB-0.1% sodium pyruvate [SP]), 23.43 (distilled water [DW]), 24.34 (phosphate buffered saline [PBS]), 24.95 (nutrient broth [NB]), 25.82 (TSB-0.6% yeast extract [YE]), and 28.27 (violet red bile glucose [VRBG]). For an inoculation of 1.82% CFU/g of Cronobacter spp. (E. sakazakii), the Ct values were recorded in this sequence: 20.34 (EE broth), 22.16 (TSB-0.6% YE), 22.37 (BHI), 22.71 (VRBL), 22.88 (TSB), 23.01 (DW), 23.19 (NB), 23.79 (TSB-0.1% SP), 24.66 (VRBG), and 24.70 (PBS). Finally, when the inoculum of Cronobacter spp. (E. sakazakii) was 0.182 CFU/g, the Ct values followed this order: 21.93 (VRBL), 23.07 (TSB-0.6% YE), 23.31 (DW), 23.47 (PBS), 23.70 (BHI), 24.14 (TSB-0.1% SP), 25.14 (TSB), 29.00 (VRBG), 31.55 (EE broth), and were undetected in the case of NB. Consequently, these results indicate that there were no significant differences among the 10 different pre-enrichment broths. Future studies should focus on exploring pre-enrichment broths that can improve the limit of detection at very low Cronobacter spp. (E. sakazakii) concentrations and enhance the selective recovery of Cronobacter spp. (E. sakazakii) under acid, antibiotic, cold, and heat damage conditions.

• 한국축산식품학회지
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• 제44권1호
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• pp.204-215
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• 2024

This study was designed to examine the effect of Lactilactobacillus curvatus LB-P9 on hair regeneration. The treatment of LB-P9 conditioned medium increased the proliferation of both hair follicle dermal papilla cells and hair germinal matrix cells (hGMCs). Moreover, the expression levels of hair growth factors such as vascular endothelial growth factor (VEGF) and fibroblast growth factor 7 were significantly elevated in hGMCs co-cultured with LB-P9. After time-synchronized depilation, mice were orally administered with either 4×10⁷ colony forming unit (CFU) of LB-P9 (low dose) or 4×10⁸ CFU of LB-P9 (high dose), once daily for 4 weeks. Compared with the vehicle (phosphate-buffered saline)-administrated group, the LB-P9-treated groups exhibited accelerated hair regrowth rate and enhanced hair thickness in a dose-dependent manner. Supporting this observation, both hair follicle numbers and the dermal thickness in skin tissues of the LB-P9-treated groups were increased, compared to those of the vehicle-treated group. These results might be explained by the increased level of β-catenin and number of hair follicle stem cells (CD34⁺ CD49f⁺ cells) in the skin tissues of mice administered with LB-P9, compared to the vehicle-treated mice. Also, increased serum levels of hair growth factors such as VEGF and insulin-like growth factor-1, and superoxide dismutase were found in the LB-P9-treated groups, compared to those of the vehicle-treated group. Taken together, these results might demonstrate that the oral administration of LB-P9 promotes hair regeneration by the enhancement of dermal papilla proliferation through the stimulation of hair growth factor production.

• Restorative Dentistry and Endodontics
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• 제49권1호
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• pp.6.1-6.16
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• 2024

Objectives: This study aimed to investigate the elemental analysis and microhardness of a bioactive material (Activa) and marginal tooth structure after storage in different media. Materials and Methods: Fifteen teeth received cervical restorations with occlusal enamel and gingival dentin margins using the tested material bonded with a universal adhesive, 5 of them on the 4 axial surfaces and the other 10 on only the 2 proximal surfaces. The first 5 teeth were sectioned into 4 restorations each, then stored in 4 different media; deionized water, Dulbecco's phosphate buffered saline (DPBS), Tris buffer, and saliva. The storage period for deionized water was 24 hours while it was 3 months for the other media. Each part was analyzed by scanning electron microscopy-energy dispersive spectroscopy (SEM-EDS) analysis for different substrates/distances and the wt% of calcium, phosphorus, silica, and fluoride were calculated. The other 10 teeth were sectioned across the restoration, stored in either Tris buffer or saliva for 24 hours or 3 months, and were evaluated for microhardness of different substrates/areas. Data were analyzed using analysis of variance and Tukey's post hoc test. Results: Enamel and dentin interfaces in the DPBS group exhibited a significant increase in calcium and phosphorus wt%. Both silica and fluoride significantly increased in tooth structure up to a distance of 75 ㎛ in the 3-month-media groups than the immediate group. Storage media did not affect the microhardness values. Conclusions: SEM-EDS analysis suggests an ion movement between Activa and tooth structure through a universal adhesive while stored in DPBS.

• Radiation Oncology Journal
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• 제28권4호
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• pp.219-223
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• 2010

목적: 국내에서 합성된 히스톤 탈아세틸 효소 억제제인 SK-7041의 생체내 방사선 감수성 증진 효과를 확인하고자 하였다. 대상 및 방법: 6주령의 웅형 C3H 마우스에 RIF-1 세포를 주입한 후 대조군 약물군 방사선군, 방사선 및 약물군으로 분류하였다. 약물군과 방사선 및 약물군에는 SK-7041 4 mg/kg을 12시간 간격으로, 6회에 걸쳐 복강 내 투여하였다. 대조군 및 방사선군에는 동일 용량의 phosphate buffered saline (PBS)를 같은 방법으로 투여하였다. 방사선군과 방사선 및 약물군에는 4번째 투약 후 6시간 후에 5 Gy를 1회 조사하였다. 이후 2~3일 간격으로 종양의 크기를 측정하여 종양 성장 연기 곡선을 산출하였다. 결과: 접종된 종양이 $1,500mm^3$까지 자라는 데 소요된 기간은 대조군, 약물군, 방사선군, 방사선 및 약물군에서 각각 10일, 9일, 9일, 12일이고, 14일째의 체적은 각각 $276.7mm^3$, $279.9mm^3$, $292.5mm^3$, $185.5mm^3$로, 각 군간의 차이는 통계학적으로 유의하였다(p=0.0004). 아울러, 대조군과 약물군의 차이와 방사선군과 방사선 및 약물군의 차이를 비교하였을 때, 통계학적으로 유의한 경향을 보였다(p=0.0650). 결론: 히스톤 탈아세틸 효소 억제제인 SK-7041 의 RIF-1 세포주에 대한 마우스내 방사선 감수성 증진 효과를 저농도에서 확인할 수 있었으며, 방사선과 SK-7041의 상호작용은 상승 작용일 가능성을 시사하였다. 향후 환자에서의 적용 가능성에 대한 연구가 필요할 것으로 생각한다.