Autophagy Enhancers Regulate Cholesterol-Induced Cytokine Secretion and Cytotoxicity in Macrophages

Objective@#Hypercholesterolaemia transforms macrophages into lipid-laden foam cells in circulation, which can activate the immune response. Compromised autophagy and inflammatory cytokines are involved in the pathogenesis and progression of metabolic diseases.The aim of this study was to identify the role of autophagy as a modulator of the inflammatory response and cytotoxicity in macrophages under hypercholesterolaemic conditions. @*Methods@#High cholesterol-induced cytokine secretion and alteration of autophagyassociated molecules were confirmed by cytokine array and western blot analysis, respectively. To confirm whether autophagic regulation affects high cholesterol-induced cytokine release and cytotoxicity, protein levels of autophagic molecules, cell viability, and cytotoxicity were measured in cultured macrophages treated autophagy enhancers. @*Results@#Cholesterol treatment increased cytokine secretion, cellular toxicity, and lactate dehydrogenase release in lipopolysaccharide (LPS)-primed macrophages. Concomitantly, altered levels of autophagy-related molecules were detected in LPS-primed macrophages under hypercholesterolaemic conditions. Treatment with autophagy enhancers reversed the secretion of cytokines, abnormally expressed autophagy-associated molecules, and cytotoxicity of LPS-primed macrophages. @*Conclusion@#Autophagy enhancers inhibit inflammatory cytokine secretion and reduce cytotoxicity under metabolic disturbances, such as hypercholesterolaemia. Modulation of autophagy may be a novel approach to control the inflammatory response observed in metabolic diseases.

Bile duct ligation of C57BL/6 mice as a model of hepatic encephalopathy

Background@#Bile duct ligation (BDL) has been used for experimental research on hepatic encephalopathy (HE) caused by chronic liver disease. However, little research has been done on a BDL model in C57BL/6 mouse. Therefore, we evaluated the suitability of a BDL model in C57BL/6 mouse for the study of HE and determined which behavioral tests are appropriate for the identification of HE in this model. @*Methods@#Twelve to fourteen-week-old male C57BL/6 mice were randomly assigned to either sham group or BDL group. Histological changes in liver were confirmed by hematoxylin/ eosin and Masson’s trichrome staining. Liver function alterations were detected by alanine aminotransferase (ALT) and ammonia levels. To identify behavioral changes, open field, elevated plus maze, novel object recognition, and passive avoidance tests were performed. @*Results@#Inflammatory liver injury and fibrosis were observed 14 days after BDL. ALT and ammonia levels were significantly higher in BDL group than in sham group. There were no differences in general locomotor activity or anxiety between the groups. No difference was observed between these two groups in the novel object recognition test, but BDL group showed significant learning/memory impairment in the passive avoidance test compared to sham group. @*Conclusions@#Fourteen days of BDL in 12–14-week-old male C57BL/6 mice is a clinically relevant model for HE, as these mice have liver fibrosis with impaired liver function, hyperammonemia, and learning/memory impairment. Passive avoidance can be used as the major behavioral test in this model of HE.

Long-Term Culture of Organotypic Hippocampal Slice from Old 3xTg-AD Mouse: An ex vivo Model of Alzheimer's Disease

OBJECTIVE: Conventional methods for organotypic hippocampal tissue slice culture (OHSC) have shown several disadvantages or limitations regarding age of animals used, duration of culture and difficulty using neurodegenerative models. Therefore, we tried to establish OHSC from old 3xTg-Alzheimer’s disease (AD) mice for longer period (over 4 weeks) and to validate utility of this system as a valid platform for translational neuroscience of AD. METHODS: OHSC was performed with old 3xTg-AD mice (12–14 months), old wild type mice (12–14 months) and young 3xTg-AD mice (2–4 months) using serum-free medium for 4 weeks. Hippocampal structure was evaluated by 4’, 6-diamidino-2-phenylindole (DAPI) intensity and neuronal metabolism was measured by Alamarblue assay. Pathologic characteristics of AD were also investigated; β-amyloid levels by ELISA, amyloid plaque deposition by Thioflavin-S staining, and glial activation by immunohistochemistry. RESULTS: Following 4-week culture in serum-free media, hippocampal cells and layers were well preserved in cultured slices from old AD mice as was in those from young AD and old wild type mice. On the contrary, excessive regression of total visible cells was observed in conventional serum-containing medium regardless of genotype of mice. In parallel with this well preserved structure, major pathologic characteristics of AD were also well manifested in hippocampal slices from old AD mice. CONCLUSION: Our findings suggest that long-term OHSC from old 3xTg-AD mouse can serve as a promising ex vivo system for studies on pathophysiology of AD, especially with the minimum number of sacrifice of experimental animals.

Hymenoptera venom anaphylaxis in adult Korean: a multicenter retrospective case study

PURPOSE: We investigated the causes, clinical features, and risk factors of bee venom anaphylaxis in Korea. METHODS: The medical records of the diagnosis of anaphylaxis during a 5-year period from the 14 hospitals in Korea have been retrospectively reviewed. Cases of bee venom anaphylaxis were identified among anaphylaxis patients, and subgroup analyses were done. RESULTS: A total of 291 patients were included. The common cause of bee species was vespid (24.6%) in bee venom anaphylaxis, followed by honeybee and vespid (8.8%), apitherapy (7.7%), and honeybee (2.0%), although the causative bee species were commonly unknown (56.9%). The severity of anaphylaxis was mostly mild-moderate (72.9%), and common clinical manifestations included cutaneous (80.6%), cardiovascular (39.2%), respiratory (38.1%), and gastrointestinal (13.1%) symptoms. Portable epinephrine auto-injectors were prescribed to 12.1% of the patients. Subject positive to both vespid and honeybee showed more severe symptoms and higher epinephrine use (P<0.05). The severity was significantly associated with older age, but not with gender, underlying allergic disease, or family history. Apitherapy-induced anaphylaxis showed a higher rate of hospitalization and epinephrine use than bee sting anaphylaxis (P<0.05). CONCLUSION: Vespid is the most common cause of bee venom anaphylaxis in Korea. It is suggested that positivity to honeybee and vespid may be associated with more severe symptoms.

The Genetic and Psychosocial Factors Affecting the Change of Drinking Behavior of Male College Students / 신경정신의학

OBJECTIVES: This study aimed to determine the general predictive factors of change in drinking behavior and to provide materials for preventing drinking problems during early adulthood through examining genetic and psychosocial factors affecting the change of drinking behavior in college students. METHODS: The subjects were 101 male college students, a part of 534 students who had completed the previous study in 2000. In the present study as a 6-years follow up, we reassessed the drinking pattern and psychosocial variables and compared the results with previous data of the same subjects. To identify factors affecting the current drinking pattern, we used stepwise multiple regression and logistic regression analysis. RESULTS: D allele (ALDH2) was found to reduce the degree of drinking and suppress problematic drinking, and C allele (TPH) had a suppression effect on problematic drinking. Drinking motive had a direct effect on the degree of drinking and problematic drinking. Negative cognitive expectancy had a direct effect on problematic drinking. CONCLUSION: Authors found some factors affecting the change of alcohol drinking behavior in college students and confirmed that there were hierarchies of significance among these factors. These may be applicable as variables for predicting drinking behavior in early adulthood.