Baicalin, a flavonoid isolated from Scutellaria baicalensis, has anti-inflammatory, antioxidant, and neuroprotective effects. Glutamate is a major neurotransmitter that plays an important role in brain function, but excessive release of glutamate causes excitotoxicity and damages cells. We investigated the neuroprotective effects of baicalin in glutamate-exposed neurons. The mouse hippocampal neuronal cell line (HT22) was cultured in a general manner, glutamate and/or baicalin were treated on the cells. Baicalin was administered 1 hr before glutamate treatment. Cells were collected 24 hr after glutamate, and cell viability was measured using MTT assay. Reactive oxygen species (ROS) and lipid peroxidation (LPO) assays were performed to measure oxidative stress. Glutamate reduced cell viability in a dose- and time-dependent manner. MTT assay showed that baicalin treatment ameliorated the decrease in cell viability due to glutamate toxicity. The effect of baicalin is dose-dependent. Glutamate caused severe nerve damage, including condensation of the cell shape, loss of dendrites and axons. However, baicalin treatment attenuated these morphological changes, and the effect of baicalin was dose-dependent. ROS and LPO analyses showed that glutamate increases oxidative stress, and baicalin attenuates this change due to glutamate toxicity. The effect of baicalin on these results was dose-dependent. We confirmed that baicalin performs an antioxidant function against glutamate toxicity in neurons. In conclusion, these results suggest that baicalin exerts neuroprotective effects on damaged neurons through antioxidant activity.

Cell culture is a widely used in vitro tool that enhances our understanding of cell biology, disease mechanisms, drug responses, and the development of tissue engineering. However, there are a number of important drawbacks to conventional two-dimensional (2D) cultures, such as the loss of polarity, altered cell shape, and disruption of cell-extracellular matrix connections. Alternatively, organoids are tissue-engineered, cell-based in vitro models derived from stem cells that can self-organize and differentiate into three-dimensional (3D) structures, recapitulating the morphology and functions of their in vivo counterparts. Bisphenol A (BPA), a ubiquitous industrial chemical, has recently gained recognition as an environmental hazard. Previous research has demonstrated that BPA negatively affects the integrity of the intestinal barrier by triggering programmed cell death and suppressing cell growth in human colonic epithelial cell lines. However, a 2D-based cellular study cannot represent its exposure to multicellular organs. This work investigates the impact of BPA on the structure and function of the intestinal barrier. We examine the effect of BPA on the proliferation and tight junction gene expression with two models: the HT-29 colon cancer cell line and an intestine organoid model and morphological changes of intestinal organoid (I/O). The proliferation was increased in a dose-dependent manner with I/O, but at the same concentration, BPA does not increase the significant number of HT-29 cell respectively. Proliferation-related gene and tight junction gene expression pattern was similar between HT-29 and I/O other than Claudin-4. Therefore, this study offers a more precise depiction of the functional and morphological alterations caused by BPA in comparison to traditional 2D cell cultures.

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.

Background: Platelet-derived growth factor receptor alpha (PDGFRα) is essential for various biological processes, including fetal Leydig cell differentiation. The PDGFRαEGFP mouse model, which expresses an eGFP fusion gene under the native Pdgfrα promoter, serves as a valuable resource for exploring PDGFRα’s expression and function in vivo. This study investigates PDGFRα expression in adult testicular cells using PDGFRαEGFP mouse model. Methods: Genotyping PCR and gel electrophoresis were used to confirm the zygosity of PDGFRαEGFP mice. Histological examination and fluorescence imaging were used to identify PDGFRα expression within testicular tissue. Immunohistochemical analysis assessed the co-expression of PDGFRα with c-Kit, ANO-1, and TASK-1 in testicular cells. Results: Genotyping confirmed the heterozygous status of the mice, which is crucial for studies due to the embryonic lethal phenotype observed in homozygotes. Histological and fluorescence imaging revealed that PDGFRα+ cells were primarily located in the interstitial spaces of the testis, specifically within Leydig cells and peritubular myoid cells (PMCs). Immunohistochemical results showed PDGFRα co-localization with c-Kit and ANO-1 in Leydig cells and a complete co-localization with TASK-1 in both Leydig cells and PMCs. Conclusions: The findings demonstrate specific expression of PDGFRα in Leydig cells and PMCs in adult testicular tissue. The co-expression of PDGFRα with c-Kit, ANO-1, and TASK-1 suggests complex regulatory mechanisms, possibly influencing testicular function and broader physiological processes.

Background: Leydig cells, crucial for testosterone production, express ion channels like ANO1 that influence hormone secretion. This study investigates the expression and role of the Tandem of P domains in a weak inward rectifying K+ channel-related Acid-Sensitive K+-1 (TASK-1) channel in these cells, exploring its impact on testicular function and steroidogenesis. Methods: TASK-1 expression in Leydig cells was confirmed using immunostaining, while RT-PCR and Western Blot (WB) validated its expression in the TM3 Leydig cell line. The effect of a TASK-1 channel blocker on cell viability was assessed through live/dead staining and MTT assays. Additionally, the blocker’s effect on testosterone secretion was evaluated by measuring testosterone levels. Results: Immunohistochemical analysis revealed a predominant presence of TASK- 1, along with c-Kit and ANO-1, in Leydig cells adjacent to seminiferous tubules and also in Sertoli and spermatogenic cells. Expression levels of TASK-1 mRNA and protein were significantly higher in TM3 Leydig cells compared to TM4 Sertoli cells. In addition, blocking TASK-1 in TM3 cells with ML365 induced cell death but did not affect LHinduced testosterone secretion. Conclusions: These findings suggest that TASK-1 in Leydig cells is crucial for their viability and proliferation, highlighting its potential importance in testicular physiology.

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.

도로 건설로 인한 서식지 파편화에 대한 저감방안으로 육교형 생태통로가 건설되고 있기는 하지만 효과성에 대해서는 아직도 논쟁이 있다. 생태통로의 효과성 평가를 위해 족적트랩, 카메라트랩과 같은 모니터링 방법이 실시되고는 있으나 얼마나 많은 개체가 이용하는지 정량적으로 평가하기에는 한계가 있다. 이에 본 연구에서는 생태통로와 인근 지역을 서식지로 이용할 가능성이 큰 소형포유류인 등줄쥐를 대상으로 포획-재포획 방법으로 개체 위치 파악을 통해 생태통로 이용 정도를 도출하고, 트랩 주변 환경 특성을 이용하여 등줄쥐의 생태통로 이용에 미치는 요인을 확인하였다. 등줄쥐의 생태통로 이용도는 격자 단위의 포획지점을 연결하여 이동 거리와 경로를 확인하였고, 생태통로 이용에 미치는 환경 특성은 트랩당 포획 횟수를 종속변수로 한 일반화 선형 모형(Generalized linear model)을 이용하였다. 연구결과, 등줄쥐의 이동 거리는 선행연구와 유사하게 나타났으며, 생태통로를 횡단하는 개체가 나타나지 않음에 따라 등줄쥐는 생태통로를 통로보다는 서식지로 이용함을 확인하였다. 등줄쥐가 생태통로를 이용하는 데 영향을 미친 환경 특성은 층위별 식생피복량(1~2m, 2~8m, 8m 이상), 교목 식생, 트랩 주변 최대 수목 흉고직경, 경사도가 유의하게 나타났다. 이에 따라 생태통로 조성 시 더 많은 교목과 관목을 식재하고, 높은 경사와 절토사면 생성을 방지하여 생태계 내 먹이원으로 이용될 수 있는 등줄쥐 이용도를 높인다면 생태통로의 효과성을 더 높일 수 있을 것으로 예상된다.

This case report describes the pathological features of pulmonary alveolar proteinosis(PAP) in mouse. Grossly, multiple irregular yellow-grey patches were observed on the lung surface of a C57BL/6 mouse. Histopathologically, the alveolar walls were thickened by type II cell hyperplasia and cellular infiltrates, and the bronchioles and alveolar lumens were filled with amorphous eosinophilic lipoproteinaceous material. This material was positive for Periodic acid-Schiff stain. Gomori methenamine silver staining was negative. Immunohistochemical staining showed that cells in the alveolar wall and lumen were positive for surfactant protein B, chitinase-like protein-3, and CD68. As a result, PAP was diagnosed. This is a rare case of spontaneous PAP in mice, and we report the histopathological characteristics along with the literature.

Macrophages secrete various cytokines and inflammatory mediators, resulting in playing critical roles in inflammation and immunity. In this study, we investigated anti-inflammatory and immune enhancing properties of PB203, which is a water-soluble extract powder from the fruit of Actinidia polygama, in macrophages. A. polygama is a medicinal plant traditionally known to treat abdominal pain, stroke and rheumatoid arthritis. However, the molecular mechanism for the immune modulation of PB203 is still unclear. Therefore, we assessed the effects of PB203 on the lipopolysaccharide (LPS)-induced inflammation and immune activation, and elucidated its action mechanism in mouse macrophage, RAW264.7 cells. PB203 significantly suppressed not only the levels of nitric oxide (NO), prostaglandin E2, tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β), but also the mRNA expression of inducible NO synthase, cyclooxygenase-2, TNF-α and IL-1β in LPS-stimulated RAW264.7 cells. We also found that these anti-inflammatory activities of PB203 were mediated through the inhibition of toll-like receptor 4 and nuclear factor kappa B (NF-κB) induced by LPS. On the other hand, in normal macrophages, PB203 dose-dependently elevated the gene expression of immunomodulators including granulocyte-macrophage colony-stimulating factor, granulocyte colony-stimulating factor, monocyte chemoattractant protein-1 and TNF-α in a statistically significant manner. The expression of IL-10, IL-1β, IL-6, and interferon-γ were also remarkably upregulated by the treatment of 500 μg/mL PB203. In addition, PB203-mediated production of NO and TNF-α was attenuated by NF-κB inhibition in RAW264.7 cells. Interestingly, PB203 promoted the production of nuclear factor erythroid-2-related factor 2, resulting in the increased level of heme oxygenase-1, which is a representative antioxidant enzyme, in both LPS-stimulated and normal RAW264.7 cells. Taken all together, these results suggest that PB203 may have great potential as the candidate of anti-inflammatory agent for improving inflammatory diseases or immune enhancing agent for preventing infectious diseases. Keywords: Actinidia polygama extract (PB203); macrophages; immunomodulator; nuclear factor kappa B (NF-κB); heme oxygenase-1 (HO-1)

목적 : 세 가지 컴퓨터 마우스 작업 시 보조기 유형(미착용, 손목보조기, 팔꿉보조기)에 따른 근활성도 차이를 비교하는 것이다. 연구방법 : 부산광역시에서 근무하는 20∼40대 사무직 근로자 36명을 대상으로 보조기 유형(미착용, 손목보조기, 팔꿉보 조기)에 따라 세 가지 컴퓨터 마우스 작업에 대한 근활성도를 비교하였다. 세 가지 마우스 작업은 과제 1(8개의 칸에 마우 스 왼쪽 버튼을 이용하여 지그재그 클릭하기), 과제 2(검지를 이용하여 6페이지 분량의 문서를 마우스 휠을 돌려서 가장 마지막 페이지로 이동하였다가 다시 되돌아 오기), 과제 3(모니터 바탕화면에 깔린 아이콘 별로 폴더에 드래그하여 옮기기 로 구성하였다. 과제 수행 중 대상자의 근활성도는 표면 근전도 장비를 사용하여 등세모근, 손가락굽힘근, 긴노쪽손목폄 근, 자쪽손목폄근을 측정하였고, 결과분석은 반복측정 분산분석을 사용하였다. 결과 : 과제 1에서 손목보조기 착용 시 긴노쪽손목폄근의 근활성도가 유의하게 감소되었고, 팔꿉보조기 착용 시에는 긴노 쪽손목폄근 및 자쪽손목폄근의 근활성도가 유의하게 감소되었다. 과제 2에서는 손목보조기 착용 시 및 팔꿉보조기 착용 시 모두 긴노쪽손목폄근과 자쪽손목폄근의 근활성도가 유의하게 감소되었다. 과제 3에서는 미착용, 손목보조기 착용, 팔 꿉보조기 착용 간 통계학적으로 유의한 근활성도 차이가 없었다. 결론 : 과제 유형에 따라 손목보조기 또는 팔꿉보조기의 착용 효과가 근육별로 다르게 나타났으며, 일부 과제에서 보조기 착용이 긴노쪽손목폄근 또는 자쪽손목폄근의 근활성도 감소에 효과가 있는 것으로 사료된다.

Haploidization in somatic cells is the process of reducing the diploid somatic chromosomes to haploid. Several studies have attempted somatic haploidization using oocytes in mice and humans. Some researchers showed partial somatic haploidization, but none observed embryo development. Our study attempted somatic haploidization using the modified somatic nuclear transfer (SCNT) protocol with various combinations of chemicals or proteins in mice. This study induced the proper segregation of somatic homologous chromosomes and full embryo development in vitro. Furthermore, somatic haploid embryos established embryonic stem cells and produced live births. The current review summarizes this recent study on the success of somatic haploidization and provides an overview of other related studies on somatic haploidization.

The purpose of this study was to investigate the effect of Cheonggukjang pills with aronia and blueberries on dextran sulfate sodium (DSS)-induced colitis in mice. There have been several reports that Cheonggukjang is effective for intestinal health, but the efficacy of Cheonggukjang containing fruits has not yet been reported. In this study, we showed the effect of cheonggukjang pills with blueberries and aronia (CPBA) on DSS-induced colitis in BALB/c mice. CPBA was obtained from Soonchang Moonokae foods and orally administered once a day for 2 weeks before DSS treatment. Colitis was induced in mice by feeding 5% (w/v) DSS drinking water for 7 days. The results showed that CPBA treatment significantly alleviated DSS-induced disease activity index associated with a decrease in colon length. CPBA improved DSS-induced histological changes and intestinal epithelial barrier integrity in mice colon. In addition, CPBA administration significantly reduced the levels of DSS-mediated interferon-γ and interleukin-6 in serum and tumor necrosis factor-α in colon tissue. Moreover, the gene expression of COX-2 and iNOS, which are factors involved in inflammatory signaling, was significantly reduced by CPBA treatment. These results suggest that CPBA have a protective effect against DSS-induced mice colitis and may be a candidate for colitis treatment.

Lipopolysaccharide (LPS) is an endotoxin factor present in the cell wall of Gram-negative bacteria and induces various immune responses to infection. Recent studies have reported that LPS induces cellular stress in various cells including oocytes and embryos. Melatonin (N-acetyl-5-methoxytryptamine) is a regulatory hormone of circadian rhythm and a powerful antioxidant. It has been known that melatonin has an effective function in scavenging oxygen free radicals and has been used as an antioxidant to reduce the cytotoxic effects induced by LPS. However, the effect of melatonin on LPS treated early embryonic development has not yet been confirmed. In this study, we cultured mouse embryos in medium supplemented with LPS or/and melatonin up to the blastocyst stage in vitro and then evaluated the developmental rate. As a result of the LPS-treatment, the rate of blastocyst development was significantly reduced compared to the control group in all the LPS groups. Next, in the melatonin only treated group, there was no statistical difference in embryonic development and no toxic effects were observed. And then we found that the treatment of melatonin improved the rates of compaction and blastocyst development of LPS-treated embryos. In addition, we showed that melatonin treatment decreased ROS levels compared to the LPS only treated group. In conclusion, we demonstrated the protective effect of melatonin on the embryonic developmental rate reduced by LPS. These results suggest a direction to improve reproduction loss that may occur due to LPS exposure and bacterial infection through the using of melatonin during in vitro culture.

Receptor tyrosine kinase c-Kit, a marker found on interstitial cells of Cajal (ICCs), is expressed in Leydig cells, which are testicular interstitial cells. The expression of other ICC markers has not yet been reported. In this study, we investigated the expression of c-Kit and anoctamin 1 (ANO1), another ICC marker, in mouse testes. In addition, the relationship between c-Kit and ANO1 expression and Leydig cell function was investigated. We observed that c-Kit and ANO1 were predominantly expressed in mouse Leydig cells. The mRNA and protein of c-Kit and ANO1 were expressed in TM3, a mouse Leydig cell line. LH induced an increase in intracellular Ca2+ concentration, membrane depolarization, and testosterone secretion, whereas these signals were inhibited in the presence of c-Kit and ANO1 inhibitors. These results show that c-Kit and ANO1 are expressed in Leydig cells and are involved in testosterone secretion. Our findings suggest that Leydig cells may act as ICCs in testosterone secretion.

This study aimed to determine whether hormonal hypersecretion could cause morphological problems in the mouse vagina and affect the ovaries and nearby extra uterine organs. All mice were synchronized to estrus before the experiment. Then human chorionic gonadotropin (hCG), progesterone, and testosterone were continuously administered for about 6 days to maintain hormone hypersecretion, and then morphological changes were analyzed, and Matrix metalloproteinases (MMPs) activity and Casp-3 expression were evaluated. As a result of the analysis, in the case of hCG, the morphological change did not show a significant difference from the vagina of normal estrus. In the case of progesterone, changes were observed in the mucosa zone and basal membrane, and it was confirmed that the activity of MMPs was increased in squamous epithelium cells. On the other hand, in the case of testosterone, overall changes in vaginal tissues were observed, and MMPs activity was increased to a very high level in all sections. The expression of Casp-3 was also the highest compared to other groups. Therefore, as a result of this study, it is thought that hormone hypersecretion affects the morphological changes of the vagina other than the ovaries and uterus and induces the activity of MMPs to cause morphological degeneration of tissues.