Cold-pressed oil from Citrus aurantifolia inhibits the proliferation of vascular smooth muscle cells via regulation of PI3K/MAPK signaling pathways.

Vascular occlusive disease is a chronic disease with significant morbidity and mortality. Although a variety of therapies and medications have been developed, the likelihood of disease re-emergence is high and this can be life-threatening. Based on a previous screening experiment related to vascular obstructive diseases using 34 types of essential oils, cold-pressed oil (CpO) from Citrus aurantifolia (lime) has been demonstrated to have the best effect for the inhibition of vascular smooth muscle cells (VSMCs) proliferation. The aim of the present study was to evaluate the effect of lime CpO on the pathological changes of VSMCs. To determine this, the effect of lime CpO on VSMC proliferation, a major cause of vascular disease, was investigated. To determine the safe concentration interval for toxicity of CpO during VSMC culture, a dilution of 1x10-5 was determined using Cell Counting Kit-8 assay, which was confirmed to be non-toxic using a lactate dehydrogenase assay. To examine the effect of lime CpO in cellular signaling pathways, changes in phosphorylation of both the PI3K/AKT/mTOR and extracellular signal-regulated MEK/ERK signaling pathways with serum were investigated. Furthermore, lime CpO with FBS also significantly decreased the expression levels of the cell cycle regulators cyclin D1 and proliferating cell nuclear antigen. Additionally, lime CpO with FBS significantly inhibited the sprouting of VSMCs in an ex vivo culture system. These results suggested that lime CpO inhibited the abnormal proliferation of VSMCs and can be developed as a nature-based therapeutic agent for obstructive vascular disease.

Alleviation effects of natural volatile organic compounds from Pinus densiflora and Chamaecyparis obtusa on systemic and pulmonary inflammation.

Chamaecyparis obtusa (C. obtusa) and Pinus densiflora (P. densiflora) have been traditionally used as antibiotic, antinociceptive and anti-inflammatory agents in Asian folk medicine. Recent studies have demonstrated antioxidant, antiproliferative and anti-inflammatory effects of C. obtusa and P. densiflora extracts. In the present study, volatile organic compounds (VOCs) of C. obtusa and P. densiflora were examined to determine whether they have anti-inflammatory capabilities. To evaluate the anti-inflammatory effects of VOCs of C. obtusa and P. densiflora, lipopolysaccharide (LPS) was administered to the lung by nasal injection and to the whole body by intraperitoneal injection. Alterations in serum immunoglobulin E (IgE) levels and prostaglandin E2 (PgE2) were examined using ELISA. LPS-increased serum IgE and PgE2 levels were recovered by administration of dexamethasone and VOCs of C. obtusa and P. densiflora. Levels of mRNA expression of inflammatory cytokines were determined in an LPS-induced inflammation mouse model. Reverse transcription-quantitative polymerase chain reaction was used to determine the mRNA expression levels of cyclooxygenase 2, interleukin (IL)-1ß, tumor necrosis factor (TNF)-α and IL-13 in peripheral blood mononuclear cells. The expression of all examined cytokine mRNAs increased by LPS was suppressed by dexamethasone and VOCs of C. obtusa and P. densiflora. Similar tendencies were observed in lung tissues and cells obtained via bronchoalveolar lavage. The results of the present study suggested that VOCs of C. obtusa and P. densiflora, through their immunosuppressive activities, may have therapeutic potential in the treatment or prevention of inflammation.

Anti-allergic Inflammatory Effects of the Essential Oil From Fruits of Zanthoxylum coreanum Nakai.

Zanthoxylum coreanum Nakai is a rare shrub which grows in Korea and China. Pericarp of Z. coreanum has been used as a crude medicine, but there are few researches about the pharmacologic activities. The present study was designed to investigate the anti-allergic inflammatory activities of the essential oil from fruits of Zanthoxylum coreanum Nakai (ZCO). Our findings showed that ZCO inhibited both the IgE-antigen complex or PMA/A23187-induced ß-hexosaminidase release and IL-4 production dose-dependently in RBL-2H3 mast cells, and confirmed that ZCO at the tested concentrations did not show cytotoxicity to RBL-2H3 cells by MTS assay. Additionally, we found that ZCO showed the significant inhibition on LPS-induced overproduction of TNF-α, IL-6 and NO. Consistently, the protein levels of iNOS and COX-2 were also remarkably decreased by ZCO treatment. Herein, Our mechanistic studies revealed that ZCO significantly suppressed the activation of transcription factor NF-κB in PMA-activated 293T cells, and further inhibited NF-κB p65 translocation into the nucleus in LPS-stimulated RAW264.7 cells. Further investigation identified that ZCO down-regulated LPS-induced phosphorylation of MAPK (JNK, ERK, and p38) signal pathway. For incremental research, we established an DNCB-induced atopic dermatitis model in BALB/c mice, and found that ZCO remarkably inhibited DNCB-induced ear swelling and AD-like symptoms. Based on these findings, ZCO is suggested to have a therapeutic potential for the allergic inflammatory diseases.

A comparative analysis of patients with mesiodenses: a clinical and radiological study.

OBJECTIVES: A mesiodens appears most commonly as a supernumerary tooth impacted in the anterior maxilla. The purpose of this study is analyze mesiodens clinically. MATERIALS AND METHODS: Gender, crown form, direction of impaction, relation to permanent incisors, and chief complaints of patients with extracted mesiodens were analyzed. RESULTS: Patients were analyzed for motivation to visit the hospital; 85.4% of the patients were referred from other hospitals. Mesiodens was more common in males than in females (3.7:1), and 70.1% of patients had only one mesiodens, while 29.6% had two mesiodenses. Of the mesiodenses, 61.4% were of the aconical form, and the most common direction was upward (62.4%), followed by the normal position (26.0%) and the horizontal position (11.6%). The mesiodenses caused orthodontic problems with the permanent incisors in 46.3% of cases. Mesiodens associated with dentigerous cyst was rarely observed in our patient group. CONCLUSION: Mesiodens is more common in males than in females and often affects the permanent incisors. Thus, careful clinical and radiological evaluations of mesiodenses are important.

Estimation of the environmental effect of natural volatile organic compounds from Chamaecyparis obtusa and their effect on atopic dermatitis-like skin lesions in mice.

Aromatherapy has been suggested as an alternative therapeutic method for the treatment of atopic dermatitis (AD), eczema and other skin diseases. In the current study, the anti-atopic properties of the volatile organic compounds of Chamaecyparis obtusa (VOCCo) were examined to determine whether they are amenable for use as a pharmaceutical candidate. The alterations in histological features, serum IgE levels and mast cell infiltration following exposure to VOCCo were determined in a 2,4-dinitrochlorobenzene (DNCB)-induced AD-like mouse model. The results of these experiments demonstrated that VOCCo inhibited the development of AD-like skin lesions by reducing the serum IgE level and mast cell infiltration into the dermal and subcutaneous layers. This was supported by screening of immune cytokine mRNAs, including interleukin (IL)-1ß and IL-6 from the skin of DNCB-treated mice. The expression of IL-1ß and IL-6 in the skin lesions of mice was dose-dependently inhibited by treatment with VOCCo. Furthermore, treatment with VOCCo resulted in the recovery of histopathological features in AD-like skin lesions. These results suggest that VOCCo may have therapeutic and preventive effects for the development of AD.

Polycationic amino acid tags enhance soluble expression of Candida antarctica lipase B in recombinant Escherichia coli.

Lipase (EC 3.1.1.3) is a popular enzyme used as an ingredient in detergents and biocatalyst in many biochemical reactions. Lipase is usually expressed in Escherichia coli as an inactive inclusion body and at a low level. In this study, Candida antarctica lipase B (CalB) was fused with various polycationic amino acid tags and expressed in E. coli in order to increase a soluble expression level. By induction with 1.0 mM IPTG, the authentic and fused CalBs were expressed at 27-56% of total protein. The 10-arginine and 10-lysine tags fused at the C-terminal of CalB significantly increased the solubility of CalB by five- to ninefold, relative to the case of the authentic CalB expressed in a recombinant E. coli Origami 2™ (DE3) strain. Among a series of the C-terminal poly-arginine tags, the recombinant CalB combined with the 10-arginine tag (CalB-R10) possessed the highest lipase specific activity of 9.5 ± 0.03 U/mg protein, corresponding to a fourfold enhancement compared with the authentic CalB.

The Aspergillus nidulans esdC (early sexual development) gene is necessary for sexual development and is controlled by veA and a heterotrimeric G protein.

The esdC (early sexual development) gene was isolated by using an expressed sequence tag (EST) as a probe from a genomic library of the early sexual developmental stage mycelia of Aspergillus nidulans. The sequence analysis revealed that the esdC gene contains a 59bp intron and encodes a 266 amino acid polypeptide with a calculated molecular weight of 29.4kDa. The EsdC protein is conserved among filamentous fungi and has a domain with similarity to a glycogen binding domain conserved in the beta subunit of the AMP-activated protein kinase (AMPK) complex. Although the esdD gene was expressed during asexual development, the expression reached its maximum at 10h and decreased thereafter up to 50h after the end of the induction of sexual development. In an esdC-null mutant under a veA(+) background, no sexual structures were formed at any condition examined. However, esdC overexpression did not lead to an induction of sexual development. In addition, to the effect of the esdC mutation on the sexual development, more conidiophores were formed in the esdC-null mutant than in a wild type. These results indicate that the esdC gene is necessary for sexual structure formation but its overexpression is not sufficient to enhance this process. Expression of the esdC gene throughout development was positively regulated by the veA gene. In addition, very little and no esdC transcript, respectively, was observed in an flbA-null mutant and in a fadA(G42R) mutant, and the esdC transcript level was higher in a fadA-null mutant and in a sfaD-null mutant than in a wild type, indicating that inactivation of FadA is necessary for positive regulation of esdC expression.

Inhibitory effects of 7-hydroxy-3-methoxy-cadalene on 4-(methylinitrosamino)-1-(3-pyridyl)-1-butanone (NNK)-induced lung tumorigenesis in A/J mice.

The present study was undertaken to estimate the effect of 7-hydroxy-3-methoxycadalene (cadalene) extracted from Zelkova serrata on 4-(methylinitrosamino)-1-(3-pyridyl)-1-butanone (NNK)-induced lung tumorigenesis in A/J mice. Twenty mice received orally NNK at a dose of 65 microg/ml water for 7 weeks following by free feeding of a commercial diet not containing cadalene for 2 weeks. Control group was maintained without NNK and cadalene administration and/or treatment groups with NNK and cadalene (6.25, 25, 100 mg/kg feed) feeding for 25 weeks. Lung tumors were induced by NNK at incidence ranging from 10 to 45%. Cadalene treatment (100 mg/kg feed) group significantly reduced the incidence of adenomas from 45 to 10% (P < 0.05), and other cadalene treatment group decreased cancer incidences in a concentration dependent manner. The results of our study strongly indicate that cadalene is capable of inhibiting development of NNK-induced lung tumorigenesis in A/J mice.