7,8-Dihydroxyflavone Attenuates Inflammatory Response and Insulin Resistance Induced by the Paracrine Interaction between Adipocytes and Macrophages.

Obesity-induced inflammation and insulin resistance are mediated by macrophage infiltration into adipose tissue. We investigated the effects of 7,8-dihydroxyflavone (7,8-DHF), a flavone found in plants, on the inflammatory response and insulin resistance induced by the interaction between adipocytes and macrophages. Hypertrophied 3T3-L1 adipocytes were cocultured with RAW 264.7 macrophages and treated with 7,8-DHF (3.12, 12.5, and 50 µM). The inflammatory cytokines and free fatty acid (FFA) release were evaluated by assay kits, and signaling pathways were determined by immunoblotting. Coculture of adipocytes and macrophages increased inflammatory mediators, such as nitric oxide (NO), monocyte chemoattractant protein-1 (MCP-1), tumor necrosis factor-alpha (TNF-α), and interleukin-6 (IL-6) and FFA secretion but suppressed the production of anti-inflammatory adiponectin. 7,8-DHF counteracted the coculture-induced changes (p < 0.001). 7,8-DHF also inhibited c-Jun N-terminal kinase (JNK) activation and blocked nuclear factor kappa B (NF-κB) nuclear translocation in the coculture system (p < 0.01). In addition, adipocytes cocultured with macrophages did not increase glucose uptake and Akt phosphorylation in response to insulin. However, 7,8-DHF treatment recovered the impaired responsiveness to insulin (p < 0.01). These findings show that 7,8-DHF alleviates inflammation and adipocyte dysfunction in the coculture of hypertrophied 3T3-L1 adipocytes and RAW 264.7 macrophages, indicating its potential as a therapeutic agent for obesity-induced insulin resistance.

Anti-Inflammatory and Anti-Diabetic Effect of Black Soybean Anthocyanins: Data from a Dual Cooperative Cellular System.

Obesity is characterized by elevated infiltration of macrophages into adipose tissue, leading to the development of insulin resistance. The black soybean seed coat is a rich source of anthocyanins with antioxidative and anti-inflammatory activities. This study investigated the effects of black soybean anthocyanin extract (BSAn) on obesity-induced oxidative stress, the inflammatory response, and insulin resistance in a coculture system of hypertrophied 3T3-L1 adipocytes and RAW264 macrophages. Coculture of adipocytes with macrophages increased the production of reactive oxygen species and inflammatory mediators and cytokines (NO, MCP-1, PGE2, TNFα, and IL-6) and the release of free fatty acids but reduced anti-inflammatory adiponectin secretion. BSAn treatment (12.5, 25, 50, and 100 µg/mL) alleviated the coculture-induced changes (p < 0.001) and inhibited coculture-induced activation of JNK and ERK signaling (p < 0.01). BSAn also blocked the migration of RAW264.7 macrophages toward 3T3-L1 adipocytes. In addition, treatment with BSAn increased PPARγ expression and glucose uptake in response to insulin in hypertrophied 3T3-L1 adipocyte and RAW264.7 macrophage coculture (p < 0.01). These results demonstrate that BSAn attenuates inflammatory responses and improves adipocyte metabolic function in the coculture of hypertrophied 3T3-L1 adipocytes and RAW264.7 macrophages, suggesting the effectiveness of BSAn for obesity-induced insulin resistance.

Predictors of blastocyst formation rate in elective day 5 transfer cycle.

This study aimed to investigate the factors affecting blastocyst formation rate. One hundred and seven fresh in vitro fertilisation (IVF) and elective day 5 blastocyst transfer cycles were selected. Univariate and multivariate analyses revealed that intracytoplasmic sperm injection (ICSI) (r = -.236, p = .014 vs. p = .005) was advantageous for blastocyst formation. In addition, the number of mature oocytes (r = -.274, p = .004 vs. p = .002) was a significant factor associated with blastocyst and good-quality blastocyst formation rates (p = .021, r = -.389). Both blastocyst and good-quality blastocyst formation rates were significantly higher with ICSI than with conventional insemination (65.0 ± 24.5% vs. 50.0 ± 21.2%, p = .012; 43.1 ± 22.8% vs. 30.9 ± 19.8%, p = .038, respectively). The number of mature oocytes appears to be the most important predictor of blastocyst formation rate. Additionally, ICSI fertilisation is superior to conventional insemination in terms of blastocyst formation rate.IMPACT STATEMENTWhat is already known on this subject? There are many advantages of blastocyst transfer cycle over cleavage transfer cycle, but there are no known routine selection criteria for the timing of embryo transfer. To date, the number of blastomeres, number of retrieved oocytes, quality of embryos and fertilisation method have been suggested as the important factors involved in blastocyst formation. However, the number of studies on this issue is limited, and some studies have shown conflicting results.What do the results of this study add? This study showed that the number of mature oocytes and ICSI fertilisation are the significant factors associated with blastocyst formation rate in elective day 5 transfer cycle.What are the implications of these findings for clinical practice and/or further research? This paper demonstrated that the number of mature oocytes and the fertilisation method should be considered before embryo transfer. Consideration of these factors would be meaningful in selecting patients who will be suitable for extended culture up to day 5.

Vitamin E: Regulatory role on gene and protein expression and metabolomics profiles.

Results from microarray analyzes have shown that both vitamin E deficiency and supplementation have a significant impact on the gene expression of various tissues and cells. Genes that were modulated by vitamin E supplementation were different depending on the tissue, which suggested that changes in gene expression are reflective of tissue function and the tissue-specific regulation of vitamin E. In addition, the magnitude of gene expression and types of genes whose expression was altered were differentially affected by the vitamin E forms used for intervention. Metabolite analyzes have provided better understanding of the vitamin E metabolic pathway and have established evidence for the regulation of energy, lipid, and glucose metabolism by vitamin E. However, there are a limited number of studies that have applied advanced genomics, proteomics, and metabolomics technologies to investigate vitamin E's biological functions and mechanisms of action. In this review, the effects of vitamin E on gene and protein expression investigated by microarray, transcriptome, and proteomics analysis are discussed. © 2019 IUBMB Life, 71(4):442-455, 2019.

Effects of mild calorie restriction on lipid metabolism and inflammation in liver and adipose tissue.

Calorie restriction (CR) has been reported to improve lipid metabolism and to decrease inflammatory diseases. However, most existing CR models use 30-50% calorie reduction, which is hard to achieve in humans. We investigated the effects of mild CR on lipid metabolism and inflammatory responses. Male C57BL/6 mice were fed control diet (10% kcal fat, Control) or high fat diet (60% kcal fat, HFD) ad libitum or reduced amount of control diet to achieve 15% CR for 16 wks. Body weights, white adipose tissue weights, liver triacylglycerol levels, and serum fetuin-A levels were lower in CR than in the Control. Serum adiponectin levels were higher in CR and lower in HFD compared with the Control. Liver and adipose tissue Mcp-1 mRNA levels were significantly lower in CR compared with the Control. Adipose tissue mRNA levels of Mcp-1, Il-6, Tnf-α and Socs3 were significantly higher in HFD than in the Control and CR, and levels of these negatively correlated with serum adiponectin levels. CR group had the lowest leptin levels and the highest liver Lepr expression, and Lepr mRNA levels positively correlated with liver Socs3 mRNA levels. Our findings showed that mild CR lowered adiposity which resulted in higher adiponectin and lower fetuin-A levels, and might have contributed to alleviation of inflammatory status in the liver and adipose tissue. Furthermore, mild CR might have affected leptin sensitivity by up-regulating Lepr expression.

Ursolic acid isolated from guava leaves inhibits inflammatory mediators and reactive oxygen species in LPS-stimulated macrophages.

Psidium guajava (guava) leaves have been frequently used for the treatment of rheumatism, fever, arthritis and other inflammatory conditions. The purpose of this study was to identify major anti-inflammatory compounds from guava leaf extract. The methanol extract and its hexane-, dichloromethane-, ethylacetate-, n-butanol- and water-soluble phases derived from guava leaves were evaluated to determine their inhibitory activity on nitric oxide (NO) production by RAW 264.7 cells stimulated with lipopolysaccharide (LPS). The methanol extract decreased NO production in a dose-dependent manner without cytotoxicity at a concentration range of 0-100 µg/mL. The n-butanol soluble phase was the most potent among the five soluble phases. Four compounds were isolated by reversed-phase HPLC from the n-butanol soluble phase and identified to be avicularin, guaijaverin, leucocyanidin and ursolic acid by their NMR spectra. Among these compounds, ursolic acid inhibited LPS-induced NO production in a dose-dependent manner without cytotoxity at a concentration range of 1-10 µM, but the other three compounds had no effect. Ursolic acid also inhibited LPS-induced prostaglandin E2 production. A western blot analysis showed that ursolic acid decreased the LPS-stimulated inducible nitric oxide synthase and cyclooxygenase protein levels. In addition, ursolic acid suppressed the production of intracellular reactive oxygen species in LPS-stimulated RAW 264.7 cells, as measured by flow cytometry. Taken together, these results identified ursolic acid as a major anti-inflammatory compound in guava leaves.

Immunoproteomically identified GBAA_0345, alkyl hydroperoxide reductase subunit C is a potential target for multivalent anthrax vaccine.

Anthrax is caused by the spore-forming bacterium Bacillus anthracis, which has been used as a weapon for bioterrorism. Although current vaccines are effective, they involve prolonged dose regimens and often cause adverse reactions. High rates of mortality associated with anthrax have made the development of an improved vaccine a top priority. To identify novel vaccine candidates, we applied an immunoproteomics approach. Using sera from convalescent guinea pigs or from human patients with anthrax, we identified 34 immunogenic proteins from the virulent B. anthracis H9401. To evaluate vaccine candidates, six were expressed as recombinant proteins and tested in vivo. Two proteins, rGBAA_0345 (alkyl hydroperoxide reductase subunit C) and rGBAA_3990 (malonyl CoA-acyl carrier protein transacylase), have afforded guinea pigs partial protection from a subsequent virulent-spore challenge. Moreover, combined vaccination with rGBAA_0345 and rPA (protective antigen) exhibited an enhanced ability to protect against anthrax mortality. Finally, we demonstrated that GBAA_0345 localizes to anthrax spores and bacilli. Our results indicate that rGBAA_0345 may be a potential component of a multivalent anthrax vaccine, as it enhances the efficacy of rPA vaccination. This is the first time that sera from patients with anthrax have been used to interrogate the proteome of virulent B. anthracis vegetative cells.

Current Status and Future Prospects of Stem Cell Therapy for Infertile Patients with Premature Ovarian Insufficiency.

Premature ovarian insufficiency (POI), also known as premature menopause or premature ovarian failure, signifies the partial or complete loss of ovarian endocrine function and fertility before 40 years of age. This condition affects approximately 1% of women of childbearing age. Although 5-10% of patients may conceive naturally, conventional infertility treatments, including assisted reproductive technology, often prove ineffective for the majority. For infertile patients with POI, oocyte donation or adoption exist, although a prevalent desire persists among them to have biological children. Stem cells, which are characterized by their undifferentiated nature, self-renewal capability, and potential to differentiate into various cell types, have emerged as promising avenues for treating POI. Stem cell therapy can potentially reverse the diminished ovarian endocrine function and restore fertility. Beyond direct POI therapy, stem cells show promise in supplementary applications such as ovarian tissue cryopreservation and tissue engineering. However, technological and ethical challenges hinder the widespread clinical application of stem cells. This review examines the current landscape of stem cell therapy for POI, underscoring the importance of comprehensive assessments that acknowledge the diversity of cell types and functions. Additionally, this review scrutinizes the limitations and prospects associated with the clinical implementation of stem cell treatments for POI.

Association of Dietary Sodium-to-Potassium Ratio with Cardiometabolic Risk Factors in Korean Adults: Findings from the Korean National Health and Nutrition Examination Survey.

High-sodium and low-potassium intakes are interdependently linked to hypertension and cardiovascular diseases. We investigated the associations of dietary sodium-to-potassium (Na/K) ratio with cardiometabolic risk factors in 12,996 Korean adults (≥30 years) from the Korean National Health and Nutrition Examination Survey Ⅶ (2016-2018). Food intake was assessed through 24 h dietary recall data. Participants were divided into thirds based on their dietary Na/K ratio, with mean molar Na/K ratios of 1.11 (low), 1.92 (medium), and 3.21 (high). Although no significant associations were found between the dietary Na/K level and the risk of hypertension, obesity, and diabetes in all participants, the high Na/K ratio group had a higher risk of hypertension compared to the low Na/K ratio group in older adults (≥65 years) after adjusting for confounding factors (odds ratio = 1.38, 95% confidence interval: 1.10-1.72). Moreover, a higher Na/K ratio was associated with an increased risk of metabolic syndrome (MetS) in all participants (p for trend = 0.0020). Within MetS components, abdominal obesity, elevated triglycerides, and elevated blood pressure were positively associated with the Na/K level. The food groups positively associated with a lower Na/K ratio were fruits, unsalted vegetables, nuts, potatoes, and dairy products. These findings suggest that a high dietary Na/K ratio may be an important risk factor for hypertension in older adults and MetS in all adults.

Development of a Nurse Turnover Prediction Model in Korea Using Machine Learning.

Nurse turnover is a critical issue in Korea, as it affects the quality of patient care and increases the financial burden on healthcare systems. To address this problem, this study aimed to develop and evaluate a machine learning-based prediction model for nurse turnover in Korea and analyze factors influencing nurse turnover. The study was conducted in two phases: building the prediction model and evaluating its performance. Three models, namely, decision tree, logistic regression, and random forest were evaluated and compared to build the nurse turnover prediction model. The importance of turnover decision factors was also analyzed. The random forest model showed the highest accuracy of 0.97. The accuracy of turnover prediction within one year was improved to 98.9% with the optimized random forest. Salary was the most important decision factor for nurse turnover. The nurse turnover prediction model developed in this study can efficiently predict nurse turnover in Korea with minimal personnel and cost through machine learning. The model can effectively manage nurse turnover in a cost-effective manner if utilized in hospitals or nursing units.

Regulation of NMDA receptor subunits after acute ethanol treatment in rat brain.

AIMS: Tolerance to ethanol-induced inhibition of N-methyl-D-aspartate receptors (NMDARs) is thought to underlie the acute adaptive mechanisms against ethanol. To explore these compensatory upregulating mechanisms of NMDARs, we investigated the expression and phosphorylation of NMDAR subunits in vivo following an acute ethanol treatment. METHODS: Male Sprague-Dawley rats were given 4 g/kg ethanol, and the phospho-S896-NR1, NR2A and NR2B subunits of NMDAR were immunoblotted from the cerebral cortex and hippocampus. We also examined the mRNAs and ubiquitinated forms of the NR2A and NR2B subunits. RESULTS: Acute ethanol treatment increased phospho-S896-NR1 at 30 min in the cerebral cortex and hippocampus, and the increase was maintained until 2 h in the hippocampus. Ethanol increased total NR2A and NR2B expression at 30 min in the cortex and hippocampus, and the NR2A increase was maintained until 2 h in the hippocampus. The increased expression of the NR2A and NR2B subunits was not associated with statistically significant alterations in mRNA expression or protein ubiquitination. CONCLUSION: Acute ethanol treatment increased NR1 subunit phosphorylation and NR2A and NR2B subunit expression in the cerebral cortex and hippocampus of rats. These effects of ethanol on the NMDAR subunits may underlie the mechanisms that compensate for ethanol-induced inhibition of NMDARs. However, the regulation of NR2A and NR2B in this paradigm is not dependent on transcriptional changes.

Perceptions and Feasibility of Actions Related to Sodium Reduction among Restaurant Owners and Cooks in Seongnam, South Korea: Comparison According to Stages of Behavioral Change.

With the increase in meals eaten outside the home, sodium reduction in restaurant foods is essential for reducing sodium intake. This study aimed to assess the stages of behavioral change for reducing sodium and the differences in perceptions among restaurant staff by stage. Restaurant owners and cooks (n = 313) in Seongnam, South Korea were surveyed on their stage of behavioral change, practices, and perceptive factors related to sodium reduction in restaurant meals using a questionnaire. The proportion of behavioral change by stage was 20.4% in the maintenance and action (MA) stage, 32.3% in the preparation (P) stage, and 47.3% in the pre-preparation (PP) stage, which included contemplation and pre-contemplation stages. The items that represent differences among the groups were recognition of social environment for sodium reduction, practice of weighing condiments and measuring salinity, and feasibility of actions related to low-sodium cooking. Logistic regression analysis was used to estimate odds ratios for practice and perceptive factors by using stage of behavioral change as the independent variable. Factors associated with being in the MA stage were weighing condiments, measuring salinity, and high feasibility of actions related to low-sodium cooking. Recognition of sodium labeling and anticipation of better taste by reducing sodium increased the odds of being in the P stage rather than the PP stage. These results suggest that customized stepwise education and support are needed for the efficacy of restaurant-based sodium reduction programs.

Psidium guajava L. leaf extract inhibits adipocyte differentiation and improves insulin sensitivity in 3T3-L1 cells.

BACKGROUND/OBJECTIVES: Psidium guajava L. (guava) leaves have been shown to exhibit hypoglycemic and antidiabetic effects in rodents. This study investigated the effects of guava leaf extract on adipogenesis, glucose uptake, and lipolysis of adipocytes to examine whether the antidiabetic properties are mediated through direct effects on adipocytes. MATERIALS/METHODS: 3T3-L1 cells were treated with 25, 50, 100 µg/mL of methanol extract from guava leaf extract (GLE) or 0.1% dimethyl sulfoxide as a control. Lipid accumulation was evaluated with Oil Red O Staining and AdipoRed assay. Immunoblotting was performed to measure the expression of adipogenic transcription factors, fatty acid synthase (FAS), and AMP-activated protein kinase (AMPK). Glucose uptake under basal or insulin-stimulated condition was measured using a glucose analog 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-D-glucose. Lipolysis from fully differentiated adipocytes was measured by free fatty acids release into the culture medium in the presence or absence of epinephrine. RESULTS: Oil Red O staining and AdipoRed assay have shown that GLE treatment reduced lipid accumulation during adipocyte differentiation. Mitotic clonal expansion, an early essential event for adipocyte differentiation, was inhibited by GLE treatment. GLE inhibited the expression of transcription factors involved in adipocyte differentiation, such as peroxisome proliferator-activated receptor γ (PPARγ), CCAAT/enhancer-binding protein α (C/EBPα), and sterol regulatory element-binding protein-1c (SREBP-1c). FAS expression was also decreased while the phosphorylation of AMPK was increased by GLE treatment. In addition, GLE increased insulin-induced glucose uptake into adipocytes. In lipid-filled mature adipocytes, GLE enhanced epinephrine-induced lipolysis but reduced basal lipolysis dose-dependently. CONCLUSIONS: The results show that GLE inhibits adipogenesis and improves adipocyte function by reducing basal lipolysis and increasing insulin-stimulated glucose uptake in adipocytes, which can be partly associated with antidiabetic effects of guava leaves.

Effect of clenbuterol on apoptosis, adipogenesis, and lipolysis in adipocytes.

Clenbuterol, a beta(2)-adrenergic receptor (beta(2)-AR) selective agonist, has been shown to decrease body fat in animals and can induce apoptosis in adipose tissue in mice. We hypothesized that direct actions of a beta-adrenergic receptor agonist on adipocytes could trigger the observed apoptotic effect. The hypothesis was inspected by investigating the direct effect of clenbuterol on apoptosis, adipogenesis, and lipolysis in vitro using the 3T3-L1 cell line and rat primary adipocytes. Cells were treated with 10(-9) to 10(-5) M clenbuterol depending on the experiments. There was no apoptotic effect of clenbuterol both in 3T3-L1 cells and rat primary adipocytes. Adipogenesis monitored by Oil Red O staining and AdipoRed assay was modestly decreased by clenbuterol treatment (p < 0.05). In fully differentiated primary adipocytes, clenbuterol increased basal lipolysis compared with the control (p < 0.01). In summary, direct stimulation of beta(2)-AR by clenbuterol does not cause apoptosis in adipocytes, despite a direct lipolytic stimulation and attenuation of adipogenesis.

Regional disparities in the associations of cardiometabolic risk factors and healthy dietary factors in Korean adults.

BACKGROUND/OBJECTIVES: Regional disparities in dietary factors might be related to regional disparities in cardiometabolic health. Therefore, this study investigated the associations of cardiometabolic risk factors and dietary factors with regional types in Korean adults. SUBJECTS/METHODS: Based on data from the 2007-2017 Korea National Health and Nutrition Examination Survey, the study included 39,781 adults aged ≥ 19 years who completed the dietary survey and a health examination. Healthy and unhealthy dietary factors (fat, sodium, fruit, and vegetable intakes) were evaluated using 1-day 24-h dietary recall method, as well as the use of nutrition labels with a questionnaire. RESULTS: Of the participants, 48.7%, 36.0%, and 15.2% lived in metropolitan, urban, and rural areas, respectively. Adults living in urban and rural had higher odds ratios (ORs) for obesity (OR for urban, 1.07; 95% confidence interval (CI), 1.01-1.14; OR for rural, 1.14; 95% CI, 1.05-1.24) than adults living in metropolitan areas; these associations were significantly observed in middle-aged adults. Compared to metropolitan residents, rural residents had lower ORs for hypertension in middle-aged (OR, 0.86; 95% CI, 0.76-0.96) and metabolic syndrome in older adults (OR, 0.78; 95% CI, 0.67-0.91). Regarding urban residents, a lower OR for diabetes in middle-aged adults (OR, 0.85; 95% CI, 0.74-0.97) and a higher OR for hypertension in older adults (OR, 1.19; 95% CI, 1.02-1.39) were observed. Overall rural residents had higher ORs of excessive carbohydrate, low fruit, and high salted-vegetable intakes than metropolitan residents. Low fruit intake was positively associated with obesity, metabolic syndrome, and hypertension, after adjustment for regional type and other confounders in total participants. CONCLUSIONS: These findings indicate that cardiometabolic risk and unhealthy dietary factors differ among regional types and age groups within Korea. Nutritional policy and interventions should consider regional types for prevention and management of cardiometabolic risk factors.

Identification of novel acetylenic alcohols and a new dihydrothiopyranone from the tropical sponge Reniochalina sp.

Two new acetylenic alcohols (1-2) and a new dihydrothiopyranone (3) were isolated from the tropical sponge Reniochalina sp. Their structures were determined by spectroscopic and chemical methods to be (3R)-hydroxyoctatriacont-(4E)-en-1-yne (1), 5-hydroxyheptatriacont-(3Z)-en-1-yne (2) and 2-hexadecyl-2,3-dihydrothiopyran-4-one (3). The acetylenic alcohol (1) exhibited significant growth inhibitory effect against human tumor cell lines.

Fucoidan from Undaria pinnatifida has anti-diabetic effects by stimulation of glucose uptake and reduction of basal lipolysis in 3T3-L1 adipocytes.

Fucoidan, a sulfated polysaccharide derived from brown seaweeds, has been shown to reduce blood glucose levels and improve insulin sensitivity in mice. We investigated the effects of fucoidan on lipid accumulation, lipolysis, and glucose uptake in 3T3-L1 cells to test the hypothesis that fucoidan exerts an anti-diabetic function by acting directly on adipocytes. The 3T3-L1 cells were treated with 10, 50, 100, and 200 µg/mL of fucoidan from Undaria pinnatifida. Oil Red O staining and AdipoRed assay were used to determine lipid accumulation during adipocyte differentiation. Fucoidan was shown to reduce lipid accumulation and glycerol-3-phosphate dehydrogenase (GPDH) activity in a dose-dependent manner (P < .01). The expression of peroxisome proliferator-activated receptor γ (PPARγ), a major transcription factor associated with adipocyte differentiation, was also suppressed upon treatment with fucoidan. Treatment with fucoidan stimulated glucose uptake in normal adipocytes and restored insulin-stimulated glucose uptake in obesity-induced insulin resistant adipocytes, which were made by incubating hypertrophied 3T3-L1 cells with the conditioned media of RAW 264.7 macrophages (RAW-CM) (P < .01). In the presence of RAW-CM, fucoidan enhanced epinephrine-stimulated lipolysis but reduced basal lipolysis, as determined by non-esterified fatty acid into the culture medium (P < .001). These results suggest that fucoidan may have anti-diabetic effects by improving insulin-stimulated glucose uptake and inhibiting basal lipolysis in adipocytes without inducing adipogenesis.

Antimicrobial activities of the bromophenols from the red alga Odonthalia corymbifera and some synthetic derivatives.

A series of bromophenols was obtained by isolation from red alga Odonthalia corymbifera and by reactions of bis(hydroxyphenyl)methanes with bromine. New bromophenols including 3,3',5,5'-tetrabromo-2,2',4,4'-tetrahydroxydiphenylmethane (10), a regioisomer of the potent antimicrobial natural product, together with known derivatives were synthesized in high yield. All of the isolated and synthesized compounds were tested for antimicrobial activity against gram-negative, gram-positive bacteria and fungi. The preliminary structure-activity relationship, to elucidate the essential structure requirements for antimicrobial activity, has been described. Among the isolated natural products 2,2',3,3'-tetrabromo-4,4',5,5'-tetrahydroxydiphenylmethane (4) was found to be the most active derivative against Candida albicans, Aspergillus fumigatus, Trichophyton rubrum, and Trichophyton mentagrophytes. The synthetic bromophenols 3,3'-dibromo-6,6'-dihydroxydiphenylmethane (13) and 3,3',5,5'-tetrabromo-6,6'-dihydroxydiphenylmethane (14) showed potent antibacterial effect against Staphylococcus aureus, Bacillus subtilis, Micrococcus luteus, Proteus vulgaris, and Salmonella typhimurium.

Enhancement of the immune responses of mice to Bacillus anthracis protective antigen by CIA07 combined with alum.

Anthrax is an acute zoonotic disease caused by infection with Bacillus anthracis. B. anthracis spores are highly resistant to environmental degradation and are used as a biological weapon. In this study, we investigated the adjuvant activity of CIA07 to anthrax protective antigen (PA). A/J mice were immunized intraperitoneally once, or twice with a 4-week interval, with recombinant PA alone or combined with alum, CpG1826, or CIA07 as adjuvant, and serum anti-PA IgG antibody responses were measured 4 weeks after each immunization. All three adjuvants significantly enhanced anti-PA IgG antibody titer 4 weeks after the priming and boosting immunizations, and alum gave the highest titer. In order to evaluate the adjuvant activity of CIA07 in the presence of alum, Balb/c mice were immunized 3 times at 1-week intervals with PA in combination with alum, CIA07 or alum plus CIA07, and the immune responses were assessed 2 weeks after the third immunization. The serum anti-PA IgG antibody titer of the CIA07-treated group was 14-fold higher than the group given PA alone, and the coadministration of CIA07 with alum further increased the titer 3.5-fold (P < 0.05). The toxin neutralizing activity of the sera from the mice given the combination of CIA07 and alum was 109-times higher than the animals given PA alone. The mice given CIA07 plus alum also showed a marked increase in the number of IFN-gamma-, IL-2-, and IL-4-producing CD4(+) T cells among their splenocytes. These data suggest the potential of CIA07 in combination with alum as an adjuvant for the development of a potent anthrax vaccine.

Effects of leptin on apoptosis and adipogenesis in 3T3-L1 adipocytes.

Leptin has been demonstrated to induce adipose tissue apoptosis, which can contribute to the decrease of adiposity, after either central nervous system or peripheral administration. However, it is not known whether leptin acts only centrally to initiate a signal or can also act directly on adipocytes to induce apoptosis. The objective of this study was to determine the direct effect of leptin on adipocyte apoptosis and adipogenesis in vitro using 3T3-L1 cell lines. An ELISA for single stranded DNA, which is highly specific for apoptotic cells, was used to quantify apoptosis. Preconfluent preadipocytes treated with 10(-9), 10(-8), 10(-7), and 10(-6)M leptin showed inhibitory effects on cell viability, and similar observations were also found in maturing preadipocytes treated during day 0-2 and day 2-4 of maturation. After 48 h incubation with 10(-6)M leptin, LDH release was increased by 24.3% (p<0.05) in preconfluent preadipocytes and by 108.5% (p<0.01) in maturing preadipocytes. However, ssDNA analysis revealed no increased apoptosis in preconfluent or maturing preadipocytes or in mature adipocytes treated with leptin. Leptin significantly reduced lipid accumulation and GPDH activity in maturing preadipocytes, demonstrating an inhibitory effect of leptin on adipogenesis. These results indicate that leptin does not act directly to induce adipocyte apoptosis, but can act directly to inhibit maturation of preadipocytes.