Amitriptyline hydrochloride (AMT), a tricyclic antidepressant, is known to exhibit antimicrobial effects against a wide range of bacterial species. This study aims to evaluate the effect of AMT on Brucella (B.) abortus infection in RAW 264.7 cells and ICR mice, which has not yet been clearly characterized. The results showed that all tested concentrations of AMT had no direct bactericidal effect on B. abortus survival at any incubation time point. Interestingly, RAW 264.7 cells pre-treated with a non-toxic high concentration of AMT before B. abortus infection showed a significant reduction in the phagocytosis of B. abortus at 20 min post-infection, compared to untreated cells. However, AMT treatment did not affect the intracellular replication of B. abortus compared to the control cells. Based on the reduced bacterial uptake observed in-vitro, an in-vivo experiment was conducted to assess whether daily oral administration of AMT at a dose of 20 mg/kg could inhibit B. abortus growth in ICR mice. The results showed that AMT treatment slightly increased both organ weights and bacterial loads, suggesting possible systemic effects of prolonged AMT exposure. In summary, these preliminary results provide initial insight into the potential effects of AMT on B. abortus infection both in-vitro and in-vivo. Therefore, further study should focus on dose optimization in-vivo and exploration of the underlying cellular mechanisms involved in AMT-mediated inhibition of phagocytosis during Brucella infection.

목적 : 본 연구에서는 회절격자가 광원의 깊이에 따라 회절영상을 주기적으로 생성하는 광학적 특성을 이용하여 중간 범위의 깊이를 측정할 수 있는 시스템을 연구하였다. 방법 : 첫 번째로 회절격자 영상시스템을 기하광학적으로 분석합니다. 두 번째로 회절격자의 주기적인 결상특성 을 파동광학적으로 분석하였다. 세 번째로 주기적인 함수사이의 콘볼루션 특성을 이용하여 깊이에 대응하는 공간주 기를 도출하였다. 네 번째로 1 m에서 4 m까지 1 m마다 타켓을 설치한후 회절격자, 카메라, 레이저로 구성된 회절 격자 영상시스템을 이용하여 회절영상을 획득하는 광학실험을 수행하였다. 다섯 번째로 첫 번째 과정에서 세 번째 과정까지 수행한 이론적 분석을 네 번째 과정에서 획득된 회절영상에 적용하여 깊이측정을 수행하였다. 마지막으로 결과 분석을 통해 시스템의 깊이해상도를 도출하였다. 결과 : 깊이 측정 실험을 수행하였고 제안된 방법의 타당성을 검증하였다. 회절영상배열의 공간주기는 타겟물체 와 회절격자사이의 거리가 멀어질수록 증가한다, 그러나 그 증가하는 비율은 물체거리에 반비례하였다. 결론 : 회절격자의 특성을 응용한 깊이 측정 시스템을 제안하였으며 이론분석과 광학실험을 통해 타당성을 검증 하였다.

The moth species and bugs are highly polyphagous that feed on a wide range of field and vegetables crops, including soybean in Korea. The monitoring study was conducted in soybean fields in Southern area Miryang from 2021-2023. Commercial funnel and rocket traps housing species-specific baiting with pheromone were used for monitoring of moth species (Spodoptera frugiperda, Spodoptera litura, Agrotis ipsilon, Helicoverpa armigera, Ostrinia furncalis, Spodoptera exigua, and Mythimna separata) and bean bugs (Riptortus pedestris), respectively. The higher number of Spodoptera species (S. litura and S. exigua) were captured in August to September with several seasonal population peaks. The higher number of R. pedestris was captured in July to September. Climatic components were found as critical and influential factors in the population dynamics of moth species and bean bug. The population information of this study can be used to understand the spring emergence along with number of generations, and integrated into management programs of moths and bugs.

The primary therapeutic approach for Brucella species infections has mainly been based on antibiotic treatment. However, the development of vaccines for brucellosis control remains controversial. Furthermore, there is currently no licensed vaccine available for human brucellosis. This study aims to evaluate the effect of a combination of recombinant protein vaccines against Brucella (B.) abortus infection using a mouse model. Two B. abortus genes, namely dapB and gpm, were cloned and expressed in competent Escherichia (E.) coli DH5α using the pCold-TF vector. Successfully cloned vectors were subjected to PCR amplification using specific primer pairs. The apparent sizes of dapB and gpm were detected at 807 bp and 621 bp, respectively. Besides, the purified recombinant proteins dapB and gpm were detected using SDS-PAGE electrophoresis with correct sizes of 82.86 kDa and 87.61 kDa, respectively. These recombinant proteins were used to immunize mice as a combined subunit vaccine (CSV) to elicit host immunity against B. abortus infection. Mice immunized with CSV exhibited increased proliferation of CD4+ and/or CD8+ T cells at week 7th and 9th before sacrifice, in comparison to the control group. Notably, CSV immunization showed a significant decrease in bacterial burden in the spleen compared to the control group. Altogether, CSV using dapB and gpm induced host adaptive immune response against Brucella infection, suggesting its potential as an effective new subunit vaccine candidate.

Spodoptera species (S. exigua and S. litura) are important pests of several crops and vegetables in Korea. We investigated development processes of Spodoptera species under constant temperatures (20, 25 and 30 oC) regimes and relative humidity (RH) (30-35, 50-55, 70-75, and 90-95%) conditions. We collected eggs of Spodoptera species by releasing them into a rectangular box inner walls covered with a sheet of white paper. Temperature and RH significantly impacted on oviposition, immature survival, adult emergence and longevity of Spodoptera species. Maximum number of eggs, shorter developmental time, higher adult emergence with longer longevity were reported in 70-75% at 30 oC. Minimal eggs and larval survival were recorded in 30-35% and 90-95% RH, respectively. This results suggest that temperature and RH had individual apparent effect on the developmental processes of Spodoptera species instead interactive effect. Therefore, there is chance to cause a significant damage to field crops and vegetables in 70-75% at 30 oC.

Subunit vaccines are being developed as a potential therapy for preventing microbial pathogen infection. In this study, the immunogenicity of recombinant Brucella (B.) abortus Fe/Mn superoxide dismutase (rFe/Mn SOD) protein as a subunit vaccine against B. abortus was investigated in BALB/c mice model. Brucella Fe/Mn SOD gene was cloned into a pcold-TF DNA vector. The bacterial recombinant protein was expressed using the Escherichia coli DH5α strain with a size of 82.50 kDa. The western blotting assay showed that rFe/Mn SOD reacted with Brucella-positive serum, indicating the potential immunoreactivity of this recombinant protein. After the second and third vaccinations, the peripheral CD4+ T cell population was increased significantly in the rFe/Mn SOD-immunized mice group compared to the PBS control group. Moreover, immunization of this recombinant protein increased the CD4+ T cell population from the first vaccination to the third vaccination. Meanwhile, the CD8+ T cells were slightly enhanced after the second vaccination compared to the first vaccination and compared to control groups. Fourteen days after the bacterial infection, the splenomegaly and the number of bacteria in the spleen were evaluated. The result showed that both rFe/Mn SOD and positive control RB51 decreased the bacterial replication in the spleen and the splenomegaly compared to control groups. Altogether, these results suggested that rFe/Mn SOD could induce host immunity against B. abortus infection.

Chlorine dioxide (ClO2) has recently emerged as an ideal disinfectant and has shown a wide range of antimicrobial activities in various pathogenic microorganisms. In this study, the virucidal effect of ClO2 at low concentration (0.02 ppm) and higher concentration (0.06 – 0.09 ppm) against Adenovirus and Herpesvirus was evaluated based on the NF T 72-281 and ASTM 1053-11 standard methods at different exposure times. The virus suspension was dried onto the carrier and then exposed to gaseous ClO2 (gClO2) at 22 ± 2∘C. For Adenovirus, exposure at a low concentration of ClO2 at the middle height resulted in the average log10 reduction of 0.95, 2.65, and 5.30 after 1, 3, and 6 h post-exposure (pe), respectively. Moreover, more than 4-log10 reduction was achieved at 4 and 6 h pe with higher concentrations of ClO2. On the other hand, the antiviral activity of gClO2 at the middle height was also effective against Herpesvirus. In particular, at 1 h pe, a less than 4-log10 reduction was observed at all examined concentrations of ClO2, whereas exposure for 3 and 6 h (with low concentration) or 2 h (with higher concentration) inactivated completely viruses attached to the carrier. These results suggested that ClO2 fumigation is a potential alternative method for disinfecting healthcare facilities, high-containment laboratories, and households with a safe concentration for human health.

We investigated the effect of a synthetic complement peptide C3a on the outcome of Brucella abortus 544 infection in a murine macrophage cell line RAW264.7 cell. First, we determined the highest non-cytotoxic concentration of the peptide in the cell line. We also found that the peptide significantly increased the growth of the bacteria at 8 and 24 h. Although the number of bacterial CFU was also elevated at 48 and 72 h, the increases were not significant as compared to controls. We further investigated the effect of C3a peptide on the growth of Brucella by pre-incubating the peptide at various temperatures and found that the effect was reversed at 24 h post-incubation suggesting that incubation of peptide at high temperatures including 65°C or 95°C could inactivate its action. This also could indicate the beneficial effect of high temperature during infection. Although several studies reported the inhibitory effect of different antimicrobial peptides including C3a, the present study preliminarily revealed that it had no positive contribution on the control of B. abortus 544 infection in vitro and indirectly to its receptor, CD88, which belongs to GPCR. Moreover, the encouraged further exploration of the effect of other similar peptides would be performed for the purpose of finding Brucella-host cell interaction for the control of disease progression.

This study aims to investigate the effects of exogenous succinic acid (SCA) on Brucella (B.) abortus infection in macrophage RAW 264.7 cells and ICR mice. Firstly, the in vitro experiment was conducted by MTT cytotoxicity and bacterial internalization assay to evaluate the uptake of B. abortus into macrophage cells. Two non-cytotoxic concentrations of SCA demonstrated attenuated invasion of Brucella into macrophages at 30 and 45 min post- infection (pi). Secondly, ICR mice were treated with SCA and infected with B. abortus. On day-14 pi, spleen and blood serum were collected to evaluate the bacterial burden and total spleen weight as well as the production of cytokine/chemokine, respectively. The results showed that SCA treatment promoted bacterial growth and reduced the total spleen weight in mice. Furthermore, SCA treatment increased the level of IL-10 cytokine in the sera, while dampening the production of MCP-1 chemokine compared to the control. The results of bacterial load in spleen and spleen weight together with cytokine/chemokine production profile in the sera indicated that SCA induced the host anti-inflammatory response which is beneficial for the survival of Brucella. Therefore, these findings suggest that SCA contributed to host immunity against Brucella infection and the emerging potential topic-immunometabolism should be invested for further investigations.

We investigated the cytotoxic potential of three different commercially available absorbent feminine hygiene products and one transdermal patch using direct contact and extract exposure methods. Two different cell lines were used – mouse fibroblast L929 and normal human skin fibroblast CCD-986sk cell lines. The test samples were extracted using three different methods in accordance to International Organization for Standardization (ISO). Viability of cells was analyzed using MTT assay and morphology of the cells were also observed using phase contrast microscopy. Overall, the direct contact method using L929 cells showed that all the test samples had no toxic effect when exposed to extracts for 1 h. For the exposure method, no toxic effect was observed in both L929 and CCD986sk cells incubated with all the test samples regardless of the extraction methods used.

This study investigated the efficacy of four Brucella (B.) abortus recombinant proteins, namely adenylate kinase (Adk), nucleoside diphosphate kinase (Ndk), 50S ribosomal protein (L7/L12) and preprotein translocase subunit (SecB), as a combined subunit vaccine (CSV) against B. abortus infection in BALB/c mice. Immunoblotting assay showed that these four recombinant proteins as well as pcold-TF vector reacted individually with Brucella-positive serum, but not with Brucella-negative serum. The peripheral blood CD4+ T cell population was increased in CSV-immunized mice compared to PBS and pcold-TF vector groups. In addition, CSV and pcold-TF groups displayed induced IgG1 and IgG2a antibodies production compared to PBS and RB51 group, whereas IgG2a titer was higher than IgG1 titer in CSV group. The secretion profiles of IgG1 and IgG2a production together with an enhancement of CD4+ T cell population suggested that CSV did not only induce T helper 1 (Th1) T cell immunity but also humoral immunity. Therein, Th1 T cell immunity is more predominant in eliminating intracellular bacteria B. abortus. Furthermore, CSV immunization significantly reduced the bacterial burden in the spleen as well as the spleen weight in comparison to PBS and pcold-TF groups. Altogether, combination of these antigens could be potential to induce protective immunity against B. abortus infection in animals.

BALB/c mice were vaccinated with Brucella (B.) abortus recombinant protein L27 (50S ribosomal protein L27) cloned into a pMal vector system. L27 was induced, purified and injected intraperitoneally (IP). Mice were vaccinated on 0-, 15- and 35-day. Serum cytokines were evaluated on 36- and 49-day from first vaccination. Mice were intraperitoneally infected with 5×104 CFU of virulent B. abortus 544 on day-50 and sacrificed after two weeks from infection. Bacterial burden from the spleen was quantified and showed a 0.7- and 0.9-log reduction in vaccinated mice in comparison to PBS and MBP (maltose binding protein) groups respectively. Cytokines in the serum demonstrated increased interferon-gamma (IFN-γ) and other pro-inflammatory cytokines such as macrophage chemoattractant protein-1 (MCP-1) and interleukin 6 (IL-6). On the other hand, interleukin 10 (IL-10) was attenuated in the sera of vaccinated mice. This cytokine profile is indicative of a cell-mediated type of immune response which is favorable for the eradication of intracellular infections. The current study showed the potential of another B. abortus ribosomal protein in inducing protective immunity against B. abortus infection.