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.

This study experimentally compares the efficacy of Situational Language Teaching (SLT) versus traditional methods in Comprehensive Chinese Language Courses (CCLC). Using an independent samples t-test for data analysis, the study confirms SLT's superior efficacy in enhancing grammatical competence and productive skills over traditional methods. The findings highlight SLT's effectiveness in creating authentic communicative contexts, fostering practical language application, and supporting skill integration. The study provides empirical evidence for adopting SLT in CCLC, particularly for grammar instruction and productive skill development, while emphasizing the importance of contextually grounded teaching practices in Chinese language education.

The role of the gut microbiota in colorectal cancer (CRC) development has garnered attention, highlighting probiotics as potential adjuncts in CRC prevention and treatment. In recent years, probiotics and their derivatives have demonstrated mechanisms that may contribute to anticancer properties. This study investigates the cytotoxic effects of Bifidobacterium bifidum KCTC 3357, Lacticaseibacillus rhamnosus KCTC 5033, Limosilactobacillus reuteri VA 103, Bacillus galactosidilyticus VA 107, and Lactococcus taiwanensis VE101 on CT-26 mouse colon carcinoma cells using live cells, heat-killed cells (paraprobiotics), and cell-free supernatants (CFS, postbiotics) through an MTT assay. The results indicate that live bacterial strains, such as KCTC 3357, VA 103, and VA 107, promoted CT-26 cell viability, while heat-killed cells and CFS exhibited dose-dependent cytotoxicity. Inactivated forms of KCTC 3357 and VE 101, as well as CFS at 10 mg/mL concentration of KCTC 5033, VA 103, and VE 101, showed the strongest antiproliferative effects. These findings suggest that non-viable probiotic derivatives, such as paraprobiotics and postbiotics, offer promising therapeutic potential for CRC, providing a safer and more stable alternative to live probiotics. However, further research is required to explore their mechanisms of action, in vivo efficacy, and potential clinical applications.

Probiotics have been evaluated as therapeutic agents for cancer treatment in an increasing number of studies. This study investigated the inhibitory and cytotoxic effects of specific Lactobacillus strains on a human colorectal adenocarcinoma cell line (HT-29). The strains assessed were Limosilactobacillus (L.) reuteri VA 102, Ligilactobacillus (L.) animalis VA 105, and Limosilactobacillus (L.) reuteri KCTC 3594 (ATCC 23272). The viability of HT-29 cells was evaluated using the MTT assay. The findings revealed that cell-free supernatants (CFS) exhibited significant anticancer effects. Heat-inactivated L. reuteri VA 105 and L. reuteri KCTC 3594 induced a pronounced reduction in cell viability. Furthermore, live cultures of L. reuteri VA 105 and L. reuteri VA 102 also showed reduced cell viability compared to the control group. These results suggest that CFS and heat-inactivated cells may be more suitable for therapeutic applications than live bacteria owing to their improved safety profiles and reduced potential for adverse effects. Our findings also emphasize the potential anticancer benefits of these LAB strains.

Probiotic lactic acid bacteria are live microorganisms that provide health benefits when administered in adequate amounts and may exhibit antiproliferative effects on various cancer cell lines, including colon cancer. This study investigates the cytotoxic effects of three Lactobacillus strains - Limosilactobacillus (L.) reuteri VA 102, Ligilactobacillus (L.) animalis VA 105, and Limosilactobacillus (L.) reuteri KCTC 3594 (ATCC 23272) - on mouse colon carcinoma cells (CT-26). Live cells, heat-killed cells, and cell-free supernatant (CFS) of Lactobacillus sp. were prepared and used to treat CT-26 cells at different concentrations. The cytotoxic effect was assessed using the MTT assay. The results indicated that the CFS of all strains significantly reduced the viability of CT-26 cells in a dose-dependent manner, with the VA 102 strain showing the most pronounced effect. Heat-killed cells of L. reuteri VA 102 and L. reuteri KCTC 3594 (ATCC 23272) also reduced cell viability. These findings suggest the potential anticancer properties of these Lactobacillus strains and indicate that CFS and heat-killed cells may offer a safer and more effective alternative to live bacteria for therapeutic applications. Our study contributes to the understanding of the potential of Lactobacillus strains, particularly L. reuteri VA 102, L. reuteri KCTC 3594 (ATCC 23272), and L. animalis VA 105, as possible candidates for cancer treatment and control.

Activated carbon (AC) is a versatile and extensively employed adsorbent in environmental remediation. It possesses distinct properties that can be enhanced to selectively target specific pollutants through modifications, including chemical impregnation or incorporation into composite materials. In this study, porous calcium alginate beads (PCAB) were synthesized by incorporating AC and natural alginate through ion gelation in a Ca(II) ion-containing solution, with the addition of sodium lauryl sulfate as a surfactant. The prepared PCAB was tested for Cu(II) removal. PCAB exhibited a spherical shape with higher porosity and surface area (160.19 m2. g−1) compared to calcium alginate beads (CAB) (0.04 m2. g−1). The adsorption kinetics followed the pseudo-first-order model for PCAB and the pseudo-second-order model for CAB. The Langmuir isotherm model provided the best fit for adsorption on PCAB, while the Freundlich model was suitable for CAB. Notably, PCAB demonstrated a maximum adsorption capacity of 75.54 mg.g−1, significantly higher than CAB's capacity of 9.16 mg. g−1. Desorption studies demonstrated that 0.1 M CaCl2 exhibited the highest efficiency (90%) in desorbing Cu(II) ions from PCAB, followed by 0.1 M HCl and 0.1 M NaCl. PCAB showed efficient reusability for up to four consecutive adsorption– desorption cycles. The fixed-bed column experiment confirmed the match with the Thomas model to the breakthrough curves with qTH of 120.12 mg.g−1 and 68.03 mg.g−1 at a flow rate of 1 mL.min−1 and 2 mL.min−1, respectively. This study indicated that PCAB could be an effective adsorbent for Cu(II) removal, offering insights for further application and design considerations.

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.

Extensive research and testing continue to be conducted for the development of vaccines targeting zoonotic diseases such as brucellosis. In this study, the potential of the DapB as a recombinant protein vaccine to effectively combat Brucella abortus 544 infection in BALB/c mice was evaluated. Western blotting assay results showed that recombinant protein DapB reacted with Brucella-positive serum, indicating its potential immunoreactivity. In vivo results showed that the peripheral blood CD4+ and CD8+ T cell population significantly increased in the DapB-immunized mice group after the first, second and third blood collection, compared to the control group that received PBS. Additionally, at the fourth blood collection, an increase in CD4+ T cell activation was observed in three vaccination groups compared to PBS negative control group. These results indicate the potential of DapB in stimulating cellular immunity. Fourteen days after infection, the bacterial load in the spleen was evaluated. The reduction in bacterial replication in the spleen by both DapB and RB51 highlights their protective efficacy against Brucella infection. These findings contribute to the ongoing efforts in developing effective vaccines against brucellosis and provide valuable insights for further research in this field.

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.