Biological and Pharmaceutical Bulletin
Online ISSN : 1347-5215
Print ISSN : 0918-6158
ISSN-L : 0918-6158
Volume 34 , Issue 8
Showing 1-36 articles out of 36 articles from the selected issue
Current Topics
  • Atsufumi Kawabata
    Type: Current Topics
    2011 Volume 34 Issue 8 Pages 1153
    Published: August 01, 2011
    Released: August 01, 2011
    JOURNALS FREE ACCESS
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  • Hiroshi Ueda
    Type: Current Topics
    2011 Volume 34 Issue 8 Pages 1154-1158
    Published: August 01, 2011
    Released: August 01, 2011
    JOURNALS FREE ACCESS
    The injury-induced intense stimulation of spinal cord neurons causes lysophosphatidic acid (LPA) biosynthesis. LPA1 receptor activation causes demyelination and sprouting of dorsal root fibers, leading to an induction of synaptic reorganization underlying allodynia, in which innocuous (tactile) stimuli cause intense pain. The LPA1 signal also initiates the up-regulation of Cavα2δ1 in dorsal root ganglion and PKCγ in the dorsal horn, underlying mechanisms for characteristic neuropathic hyperalgesia in myelinated sensory (A-type) fibers. On the other hand, the LPA3 receptor mediates microglia activation at the early stage after nerve injury and LPA-induced LPA biosynthesis. Thus, both the LPA1 and LPA3 receptors play key roles in the initiation step using a feed-forward system for neuropathic pain.
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  • Makoto Tsuda, Hidetoshi Tozaki-Saitoh, Kazuhide Inoue
    Type: Current Topics
    2011 Volume 34 Issue 8 Pages 1159-1162
    Published: August 01, 2011
    Released: August 01, 2011
    JOURNALS FREE ACCESS
    Platelet-activating factor (PAF) is a phospholipid mediator that regulates the functions of a variety of cells in the peripheral tissues and in the nervous system. Findings that injection of PAF exogenously at the skin or in the spinal cord induced pain hypersensitivity gave us much attention to its role in pain signaling. Studies using pharmacological and genetic tools to control the functions of the PAF receptor (PAFR) revealed that the PAF/PAFR system plays a role in tissue injury-induced pain, but not in the acute physiological pain evoked by thermal and mechanical stimuli. Recent investigations have focused on the roles of PAFR in pathological chronic pain such as the neuropathic pain that occurs after nerve injury for which there is currently no effective therapy. Nerve injury upregulated PAFRs in dorsal root ganglion (DRG) neurons. Studies using PAFR antagonists and PAFR-deficient mice indicated a crucial role of PAFR in production of tumor necrosis factor α (TNFα) and interleukin-1β (IL-1β) in the DRG and in developing and maintaining neuropathic pain. Thus, blocking PAFRs may be a viable therapeutic strategy for treating neuropathic pain.
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  • Koichi Noguchi, Masamichi Okubo
    Type: Current Topics
    2011 Volume 34 Issue 8 Pages 1163-1169
    Published: August 01, 2011
    Released: August 01, 2011
    JOURNALS FREE ACCESS
    The purpose of this review is to summarize the recent studies examining the expression of leukotrienes (LTs) and their receptors in nociceptive pathways, and their crucial roles in pathological pain conditions. LTs belong to a large family of lipid mediators, termed eicosanoids, which are derived from arachidonic acids and released from the cell membrane by phospholipases. LTs are known to be important factors in a variety of local and systemic diseases and allergic/inflammatory diseases. We examined whether LTs were implicated in neuropathic pain following peripheral nerve injury. Using the SNI model in rats, we investigated the expression of LT synthases and receptors mRNAs in the spinal cord and the roles on the pain behaviors. We found the expression of 5-lipoxygenase (5-LO), FLAP and the cysteinyl leukotrienes (CysLT1) mRNAs in spinal microglia, LTA4h and LTC4s mRNAs in both spinal neurons and microglia, and BLT1 mRNA in spinal neurons. Administration of the 5-LO inhibitor or the receptor antagonists suppressed mechanical allodynia. Our findings suggest that the increase of LT synthesis in spinal microglia produced via p38 mitogen-activated protein kinase (MAPK) plays a role in the generation of neuropathic pain. We also examined the expression and roles on pain behaviors of LT receptors in the dorsal root ganglion (DRG) using a peripheral inflammation model. The data indicate CysLT2 expressed in DRG neurons may play a role as a modulator of P2X3, and contribute to the potentiation of the neuronal activity following peripheral inflammation. This review summarizes the hypothesis that LTs might work in the spinal cord and primary afferent in pathological pain conditions.
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  • Atsufumi Kawabata
    Type: Current Topics
    2011 Volume 34 Issue 8 Pages 1170-1173
    Published: August 01, 2011
    Released: August 01, 2011
    JOURNALS FREE ACCESS
    Prostaglandin E2 (PGE2), a cyclooxygenase (COX) product, is the best known lipid mediator that contributes to inflammatory pain. Nonsteroidal anti-inflammatory drugs (NSAIDs), inhibitors of COX-1 and/or COX-2, suppress inflammatory pain by reducing generation of prostanoids, mainly PGE2, while they exhibit gastrointestinal, renal and cardiovascular toxicities. Selective inhibitors of microsomal PGE synthase-1 and subtype-selective antagonists of PGE2 receptors, particularly EP1 and EP4, may be useful as analgesics with minimized side-effects. Protein kinase C (PKC) and PKA downstream of EP1 and EP4, respectively, sensitize/activate multiple molecules including transient receptor potential vanilloid-1 (TRPV1) channels, purinergic P2X3 receptors, and voltage-gated calcium or sodium channels in nociceptors, leading to hyperalgesia. PGE2 is also implicated in neuropathic and visceral pain and in migraine. Thus, PGE2 has a great impact on pain signals, and pharmacological intervention in upstream and downstream signals of PGE2 may serve as novel therapeutic strategies for the treatment of intractable pain.
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  • Shogo Tokuyama, Kazuo Nakamoto
    Type: Current Topics
    2011 Volume 34 Issue 8 Pages 1174-1178
    Published: August 01, 2011
    Released: August 01, 2011
    JOURNALS FREE ACCESS
    Fatty acids, which are the essential nutrients for humans, are an important source of energy and an essential component of cell membranes. They also function as signal transduction molecules in a range of biological phenomena. Recently, an increasing number of physiologic and pharmacologic reports on fatty acids have improved our understanding of the association of fatty acids with certain diseases. It has also become apparent that functional properties of fatty acids are modulated by factors such as the amount of individual fatty acid intake and their distribution among organs. Recently, the functional relationship between polyunsaturated fatty acids and pain has been the focus of many studies. Both basic and clinical studies have shown that a dietary intake of n-3 series polyunsaturated fatty acids results in a reduction in the pain associated with rheumatoid arthritis, dysmenorrhea, inflammatory bowl disease, and neuropathy. In addition, levels of n-6 series polyunsaturated fatty acids are high in patients with chronic pain. These results indicate that polyunsaturated fatty acids play a vital role in pain regulation. In this review, we summarize a number of basic and clinical studies on polyunsaturated fatty acids and their association with pain.
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Regular Articles
  • Yi-Dong Yan, Jung Ae Kim, Mi Kyung Kwak, Bong Kyu Yoo, Chul Soon Yong, ...
    Type: Regular Article
    2011 Volume 34 Issue 8 Pages 1179-1186
    Published: August 01, 2011
    Released: August 01, 2011
    JOURNALS FREE ACCESS
    In this study, a novel liquid self-emulsifying drug delivery system (SEDDS) containing curcumin was formulated and further developed into a solid form by a spray drying method using Aerosil 200 as the solid carrier. The optimum liquid SEDDS consisted of Lauroglycol Fcc, Labrasol and Transcutol HP as the oil phase, the surfactant and the co-surfactant at a weight ratio of 15.0 : 70.8 : 14.2 (w/w/w), respectively. There was no difference in droplet size between the emulsions obtained from the liquid and solid forms of SEDDS. Solid state characterization of the solid SEDDS was performed by scanning electron micrograph (SEM), differential scanning calorimetry (DSC), and X-ray powder diffraction (XRPD). The drug formulated in the solid SEDDS was quickly and completely dissolved within 5 min, both in 0.1 N HCl and phosphate buffer pH 6.8 dissolution media, whereas crude curcumin powder was significantly less dissoluble. The solid SEDDS formulation was stable for at least 3 months at 40°C with 75% relative humidity. After oral administration to rats, curcumin in the solid SEDDS resulted in significant improvement in in vivo absorption compared with that of curcumin powder. As the dose of curcumin formulated in solid SEDDS increased from 25 to 100 mg/kg, the Cmax and area under the drug concentration time curve (AUC) of curcumin were increased by 4.6 and 7.6 times, respectively. However, the over-proportional increase in the AUC in the higher dose group might be due to underestimation of AUC in the lower dose group. In conclusion, this solid SEDDS is a promising solid dosage form for poorly water-soluble curcumin.
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  • Yukari Tanaka, Tomoyuki Ohkawa, Hiroyuki Yasui
    Type: Regular Article
    2011 Volume 34 Issue 8 Pages 1187-1193
    Published: August 01, 2011
    Released: August 01, 2011
    JOURNALS FREE ACCESS
    In this study, we developed a novel methodology, multiple injection method (MIM), for higher-throughput screening of compounds by liquid chromatography-tandem mass spectrometry (LC-MS/MS). MIM involves continuous injections of multiple samples containing a different compound respectively into the column, and then temporarily trapping of analytes at the column head in high-pressure liquid chromatography (HPLC) system. This is followed by elution of all the compounds from the column and detection of them by MS/MS. In this study, fexofenadine, verapamil, risperidone, ondansetron, and imipramine were used as model compounds to investigate the effectiveness of MIM in pharmacokinetic studies. Analytical time of validation samples of these model compounds could be shortened to one third by MIM, compared with the conventional method. In addition, both the accuracy and precision of MIM met the general criteria for quantitative analysis. The peak intensity was found to be unaffected by overlapping compounds even if they have wide range of ionization efficiency. As a result of the comparison of MIM and conventional method in the analysis of samples in pharmacokinetic studies using model compounds, no difference was shown in the quantification values. Consequently, this method has some advantages, reduction of analytical time, the improvement of sensitivity, and the simplicity of system, compared to the conventional methods. MIM should be very useful and powerful method for drug development without an additional hardware and can be used for the measurement of compounds in biological samples for pharmacokinetic studies, especially it greatly contributes to accelerating drug development in its discovery stages.
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  • Hong Jiang, Xinying Tian, Yansu Guo, Weisong Duan, Hui Bu, Chunyan Li
    Type: Regular Article
    2011 Volume 34 Issue 8 Pages 1194-1197
    Published: August 01, 2011
    Released: August 01, 2011
    JOURNALS FREE ACCESS
    Oxidative damage plays a critical role in many neurodegenerative diseases. Astrocytes are involved in supporting the survival and protection of neurons against oxidative damage. The dysfunction of antioxidant in astrocytes has been implicated in a variety of neurodegenerative disorders, such as amyotrophic lateral sclerosis (ALS), spinalmuscularatrophy (SMA). The loss of motor neuron in spinal cord has been attributed to deterioration of astrocytes. The activation of antioxidantive function in astrocytes may serve as a therapeutic strategy for neurodegenerative diseases. Nuclear factor erythroid 2-related factor 2 (Nrf2) is a master transcriptional regulator of phase II antioxidantive genes. We report herein that curcumin significantly activates Nrf2 target genes in primary spinal cord astrocytes, decreases the level of intracellular reactive oxygen species (ROS), and attenuates oxidative damage and mitochondrial dysfunction.
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  • Xiaorong Guo, Xiaoguo Wang, Wenhua Su, Guangfei Zhang, Rui Zhou
    Type: Regular Article
    2011 Volume 34 Issue 8 Pages 1198-1203
    Published: August 01, 2011
    Released: August 01, 2011
    JOURNALS FREE ACCESS
    Scutellaria baicalensis GEORGI (Lamiaceae) is the botanical origin of the well-known traditional Chinese medicine “Huang Qin” (Radix Scutellariae). Due to overexploitation that had induced a decline in natural sources, the dried roots of its congeners, S. amoena, S. rehderiana, and S. viscidula, have been used to adulterate it in recent years. This practice may cause a series of inconsistent therapeutic effects and quality control problems in the herbal medicine industry. Hence, we sequenced and analyzed three candidate DNA barcodes, the ribosomal RNA maturase gene (matK), the ribulose-1,4-bisphosphate carboxylase large subunit gene (rbcL), and the psbA–trnH intergenic spacer (psbA–trnH), to discriminate S. baicalensis and its adulterants. All candidate DNA barcodes had been successfully amplified from leaf samples. Comparatively, only psbA–trnH had been yielded from commercially prepared crude drug samples. Based on the sequence divergence, rbcL can assign S. baicalensis and its adulterants into the correct family and genus, whereas, either matK or psbA–trnH can accurately discriminate S. baicalensis and its adulterants. We proposed the multilocus barcodes rbcL+psbA–trnH for the species identification of S. baicalensis and its adulterants, and the unique barcode psbA–trnH for the authentication of commercial Radix Scutellariae. The DNA barcoding technique could be applied to the quality control of “Huang Qin”-based medicinal preparations and to the management of medicinal herb trade in the markets.
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  • Maria Grazia Cerrito, Alessandra Scagliarini, Alberto Froio, Angela Li ...
    Type: Regular Article
    2011 Volume 34 Issue 8 Pages 1204-1214
    Published: August 01, 2011
    Released: August 01, 2011
    JOURNALS FREE ACCESS
    Heme oxygenase-1 (HO-1, encoded by the HMOX1 gene) and inducible nitric oxide synthase (iNOS) have been implicated in vascular disease; however the role of these genes remains unclear. Therefore, we studied the mechanism by which iNOS-derived nitric oxide (NO) affects the intimal hyperplasia (IH) formation in relation to HO-1. We show, in a model of balloon injury in rats, that the suppression of vascular smooth muscle cells (VSMC) proliferation by NO required HO-1, while induction of apoptosis of the VSMC by NO does not involve HO-1. To better clarify the molecular mechanism of this finding, we used Hmox1+/+ and Hmox1−/− VSMC exposed to NO. In Hmox1+/+ VSMC, NO is antiproliferative (up to 34% inhibition) and it is associated to an increase of apoptosis (up to 35%) due to a decrease of X-linked inhibitor of apoptosis protein (XIAP) expression level and to the activation of caspase-3. In the absence of HO-1 (Hmox1−/− VSMC) apoptosis was significantly greater (69% p<0.01 vs. Hmox1+/+ VSMC) demonstrating that HO-1 attenuated the pro-apoptotic effect of NO on VSMC. In the context of IH, the pro-apoptotic effect of NO on VSMC is increased in the absence of HO-1 and exerts therapeutic effects with a significant reduction in IH.
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  • Yoshihiro Tokudome, Atsuo Ito, Makoto Otsuka
    Type: Regular Article
    2011 Volume 34 Issue 8 Pages 1215-1218
    Published: August 01, 2011
    Released: August 01, 2011
    JOURNALS FREE ACCESS
    Zinc-containing β-tricalcium phosphate (ZnTCP) nano particles were injected into zinc-deficient rats to promote osteogenesis. Sprague-Dawley (SD) rats (4 weeks old, average weight of 70 g) were divided into four groups: Normal rats (not ovariectomized (OVX)), Control rats (OVX), and OVX rats injected with a suspension of ZnTCP nano particles or ZnSO4. The ZnTCP contained 6.17% zinc. The suspensions (0.6 mg as a zinc volume/0.2 ml) were injected around the jaw bone once a week for 12 weeks. Local effects on the bone mineral content (BMC) of jawbone, and systemic effects on body weight, the BMC of both femurs determined by X-ray computed tomography, and bone mechanical strength (BMS) measured by the three-point bending method, were examined. The BMC of jaw bone was significantly higher in the ZnTCP-treated group than un-treated or ZnSO4-treated group. Body weight, the BMC of femurs, and BMS were also significantly higher in the ZnTCP treated-group. The zinc-containing β-tricalcium phosphate nano particles were effective at preventing bone loss induced by ovariectomy in rats and have potential uses for treating periodontitis.
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  • Dongyuan He, Li Lee, Junwei Yang, Xiaoyun Wang
    Type: Regular Article
    2011 Volume 34 Issue 8 Pages 1219-1226
    Published: August 01, 2011
    Released: August 01, 2011
    JOURNALS FREE ACCESS
    Renal interstitial fibrosis is a common outcome of a variety of chronic renal diseases. Here we evaluated the therapeutic efficacy of rhein on renal interstitial fibrosis induced by unilateral ureteral obstruction (UUO) and investigated the potential mechanisms. Mice underwent UUO, followed by orally administrated rhein (150 mg/kg/d) or control vehicle. Renal interstitial injury and the degree of fibrosis were evaluated by pathological staining and Western blot. The possible mechanisms were studied by Western blot, indirect immune-fluorescence and enzyme-linked immunosorbent assay. Our results showed that rhein therapy markedly ameliorated renal interstitial fibrotic lesions, reduced α-smooth muscle actin (α-SMA) expression, attenuated deposition of fibronectin (FN). Rhein also suppressed transforming growth factor-β1 (TGF-β1) and its type I receptor expression in obstructed kidneys. In vitro, rhein abolished the α-SMA and fibronectin expression of rat kidney interstitial fibroblasts cells (NRK-49F) induced by TGF-β1. These observations strongly suggest that rhein is a potent inhibitor of renal interstitial fibrosis, and its therapeutic mechanism is, at least in part, blocking interstitial fibroblasts cells activation.
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  • Akiko Obinata, Yoshihiro Akimoto
    Type: Regular Article
    2011 Volume 34 Issue 8 Pages 1227-1230
    Published: August 01, 2011
    Released: August 01, 2011
    JOURNALS FREE ACCESS
    We showed previously that transdifferentiation of skin epidermis to mucous epithelium can be induced by treatment with 20 μM retinol for 1 d followed by culture for 4 d without retinol in chick embryonic tarsometatarsal skin. In mouse epidermal cells, 3 μM retinoic acid (an active metabolite of retinol) inhibits epidermal keratinization in consistent with an increase in transglutaminase (TG)2/Gh, while its physiological role in the skin is still unresolved. TG1, TG3 and TG5 are also found in mammalian keratinocytes and play an important role in the formation of the stratum corneum in the skin by the introduction of cross-links into proteins. The most characteristic enzyme function of TG family is calcium-dependent transamidation activity (transamidase) that introduces inter or intramolecular ε-(γ-glutamyl)lysine cross-links into the protein. TG2/Gh is a multifunctional protein and ubiquitously expressed member of transglutaminase family that has been implicated in a variety of biological processes. By in situ hybridization analysis, we showed that TG2/Gh mRNA expression started to increase throughout the skin during the culture for 1 d with retinol, while it was weak in the control skin. On the other hand, an expression of TG3 mRNA was increased in the keratinized epidermis of control skin but was decreased by retinol. In situ transamidase activity of transglutaminase was weak in retinol-pretreated skin. Therefore, it was indicated that functions other than transamidase of TG2/Gh protein might be important in retinol-induced epidermal mucous transdifferentiation.
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  • Dan Shu, Yong Qing, Qingyi Tong, Yang He, Zhihua Xing, Yinglan Zhao, Y ...
    Type: Regular Article
    2011 Volume 34 Issue 8 Pages 1231-1239
    Published: August 01, 2011
    Released: August 01, 2011
    JOURNALS FREE ACCESS
    Deltonin is an active component purified from Dioscorea zingiberensis WRIGHT (DZW), and has shown anticancer effects. However, its mechanism of action remains elusive. In the present study, we investigated the effect of Deltonin on a panel of cancer cell lines and analyzed its mechanism in C26 cells, a murine colon carcinoma cell. Our results showed that Deltonin markedly inhibited the growth of all examined cancer cell lines. Deltonin induced dose- and time-dependent apoptosis in C26 cells. The event of apoptosis was accompanied by the release of cytochrome c, depolarization of mitochondrial membrane potential, and dose- and time-dependent reactive oxygen species (ROS) generation. Deltonin also increased the expression of Bax, decreased the expression of B-cell lymphoma/lewkmia-2 (Bcl-2), and induced the activation of caspase 9, caspase 3 and poly(ADP-ribose) polymerase (PARP). Furthermore, Deltonin decreased Akt and extracellular signal-regulated kinase-1/2 (ERK1/2) activity. These results demonstrate that Deltonin mediates the growth inhibition of cancer cells through multiple targets, which include the generation of reactive oxygen species (ROS), mitochondrial apoptosis and the inhibition of the mitogen-activated protein kinase (MAPK) and Akt signaling pathways, suggesting Deltonin is a potent cancer preventive and therapeutic agent.
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  • Ying Zhou, Pei-Hong Sun, Yu-Wang Liu, Xia Zhao, Lei Meng, Yi-Min Cui
    Type: Regular Article
    2011 Volume 34 Issue 8 Pages 1240-1245
    Published: August 01, 2011
    Released: August 01, 2011
    JOURNALS FREE ACCESS
    The objectives of the study were to assess the safety and pharmacokinetics of silodosin capsules in 82 healthy male Chinese subjects. To evaluate the safety after single-dosing escalation, 40 subjects were equally divided into 4 groups (2, 4, 8, 12 mg) by a randomized, double-blind and placebo-controlled design. To assess the pharmacokinetics after single-dosing, 30 subjects were equally divided into 3 groups (4, 8, 12 mg). To assess the safety and pharmacokinetics via multiple-dosing, 12 subjects were included as a group (4 mg once daily at day 1 and day 7; 4 mg twice daily at day 2 through day 6). The safety observations showed that mild adverse events, including postural hypotension, dizziness, and headache, were observed. After single-dosing at doses of 4, 8, and 12 mg, the mean area under the concentration–time curve from 0 to 36 h (AUC0—36) values were 136.82±46.38, 270.17±54.66, and 474.63±108.50 μg/l·h and the mean maximal silodosin concentration in plasma (Cmax) values were 26.70±7.48, 48.47±12.35, and 94.07±22.59 μg/l, respectively. After multiple-dosing, the Cmax value at day 7 was 33.84±19.54 μg/l, and the AUC0—24 value at day 7 was 193.19±68.96 μg/l·h. The accumulation ratio of the AUC value was 1.55 by comparing the multiple-dosing with the single-dosing. It is concluded that silodosin is safe and tolerated in healthy Chinese male subjects at the dosing levels used in this study. The mean Cmax and AUC values of silodosin increased proportionally with dose escalation, showing characteristics of linear pharmacokinetics.
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  • Kazunori Iwanaga, Shinji Yoneda, Yukimi Hamahata, Makoto Miyazaki, Mak ...
    Type: Regular Article
    2011 Volume 34 Issue 8 Pages 1246-1251
    Published: August 01, 2011
    Released: August 01, 2011
    JOURNALS FREE ACCESS
    Furanocoumarin derivatives, known as components of grapefruit juice, showing inhibitory effects against P-glycoprotein (P-gp) in the intestine are also contained in the plants of rutaceae and umbelliferae families, which are used as components of Kampo extract medicines. In this study, we investigated the inhibitory effects of byakangelicol and rivulobirin A, known as furanocoumarins showing P-gp inhibitory effect using Caco-2 monolayer, against P-gp at the blood–brain barrier (BBB) under both in vitro and in vivo conditions. First we studied the membrane permeability of furanocoumarins to clarify whether they can be absorbed from the intestine. Both furanocoumarins showed high permeability through the Caco-2 monolayer, suggesting that they can easily reach the systemic circulation after oral administration. Then, we evaluated the effect of these furanocoumarins on the uptake of calcein acetoxymethyl ester (calcein-AM), a P-gp substrate, into bovine brain microvascular endothelial cells (BBMEC). Both furanocoumarins significantly increased the uptake amount of calcein-AM into BBMEC by the inhibition of P-gp at the BBB in vitro. Next we also investigated the P-gp inhibitory effect of these furanocoumarins at the rat BBB in vivo using verapamil as a P-gp substrate. Both furanocoumarins increased the B/P ratio of verapamil compared to the control, even under in vivo conditions; however, the extent of the inhibitory effect was much lower than in vitro condition. In conclusion, byakangelicol and rivulobirin A may inhibit P-gp expressed at the BBB even under in vivo conditions. Further studies using Kampo extract medicines under in vivo condition are necessary for safe drug therapy.
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  • Shuso Takeda, Akari Hirayama, Shino Urata, Nobutaka Mano, Keiko Fukaga ...
    Type: Regular Article
    2011 Volume 34 Issue 8 Pages 1252-1256
    Published: August 01, 2011
    Released: August 01, 2011
    JOURNALS FREE ACCESS
    15-Lipoxygenase (15-LOX) is one of the key enzymes responsible for the formation of oxidized low-density lipoprotein (ox-LDL), a major causal factor for atherosclerosis. Both enzymatic (15-LOX) and non-enzymatic (Cu2+) mechanisms have been proposed for the production of ox-LDL. We have recently reported that cannabidiol-2′,6′-dimethyl ether (CBDD) is a selective and potent inhibitor of 15-LOX-catalyzed linoleic acid oxygenation (Takeda et al., Drug Metab. Dispos., 37, 1733—1737 (2009)). In the LDL, linoleic acid is present as cholesteryl linoleate, the major fatty acid esterified to cholesterol, and is susceptible to oxidative modification by 15-LOX or Cu2+. In this investigation, we examined the efficacy of CBDD on i) 15-LOX-catalyzed oxygenation of cholesteryl linoleate, and ii) ox-LDL formation catalyzed by 15-LOX versus Cu2+-mediated non-enzymatic generation of this important mediator. The results obtained demonstrate that CBDD is a potent and selective inhibitor of ox-LDL formation generated by the 15-LOX pathway. These studies establish CBDD as both an important experimental tool for characterizing 15-LOX-mediated ox-LDL formation, and as a potentially useful therapeutic agent for treatment of atherosclerosis.
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  • Takahiro Hayashi, Yuriko Nozaki, Makoto Nishizuka, Masahito Ikawa, Shi ...
    Type: Regular Article
    2011 Volume 34 Issue 8 Pages 1257-1263
    Published: August 01, 2011
    Released: August 01, 2011
    JOURNALS FREE ACCESS
    To clarify the molecular mechanism of adipocyte differentiation, we previously isolated a novel gene, factor for adipocyte differentiation (fad) 158, whose expression was induced during the earliest stages of adipogenesis, and its product was localized to the endoplasmic reticulum. We found that the knockdown of fad158 expression prevented the differentiation of 3T3-L1 cells into adipocytes. In addition, over-expression of fad158 promoted the differentiation of NIH-3T3 cells, which do not usually differentiate into adipocytes. Although these findings strongly suggest that fad158 has a crucial role in regulating adipocyte differentiation, the physiological role of the gene is still unclear. In this study, we generated mice in which fad158 expression was deleted. The fad158-deficient mice did not show remarkable changes in body weight or the weight of white adipose tissue on a chow diet, but had significantly lower body weights and fat mass than wild-type mice when fed a high-fat diet. Furthermore, although the disruption of fad158 did not influence insulin sensitivity on the chow diet, it improved insulin resistance induced by the high-fat diet. These results indicate that fad158 is a key factor in the development of obesity and insulin resistance caused by a high-fat diet.
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  • Akihisa Abe, Hiroyuki Yamada, Shota Moriya, Keisuke Miyazawa
    Type: Regular Article
    2011 Volume 34 Issue 8 Pages 1264-1272
    Published: August 01, 2011
    Released: August 01, 2011
    JOURNALS FREE ACCESS
    β-Carboline alkaloids are naturally occurring plant substances that have a wide spectrum of neuropharmacological, psychopharmacological, and antitumor effects. Recently, we have demonstrated that harmol, a β-carboline alkaloid, induces apoptosis by caspase-8 activation independently from Fas/Fas ligand interaction in human non-small cell lung cancer (NSCLC) H596 cells. Here, we found that harmol induces autophagy and cell death in human NSCLC A549 cells. Although harmol induced cell death in A549 cells in a significant dose- and time-dependent manner, it did not induce caspase-3, caspase-8, or caspase-9 activity. Furthermore, cleavage of poly-(ADP-ribose)-polymerase was not induced in A549 cells by harmol treatment. Autophagy, but not apoptosis, was detected by electron microscopy in A549 cells treated with 70 μM harmol. Pretreatment of A549 cells with 3-methyladenine, an autophagy inhibitor, as well as small interfering RNA (siRNA)-mediated knockdown of LC3, both suppressed harmol-induced cell death. These suggest that the induction of autophagy by harmol precedes cell death. The cytotoxicity of some anticancer agents is reportedly linked to autophagy induction. The 2 major autophagy regulatory pathways are the Akt/mammalian target of rapamycin (mTOR) pathway and the extracellular signal-regulated kinase 1/2 (ERK1/2) pathway. Although harmol treatment showed no effect on the Akt/mTOR pathway, it transiently activated the ERK1/2 pathway. However, inhibition of the ERK1/2 pathway using the mitogen-activated protein kinase (MEK)/ERK inhibitor U0126 partially suppressed autophagy. Therefore, although activation of the ERK1/2 pathway might be related to harmol-induced autophagy, another major pathway may also be involved in A549 cells.
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  • Tetsuya Yoshino, Tomoyoshi Ishikawa, Takashi Ishihara, Yoshimi Takeuch ...
    Type: Regular Article
    2011 Volume 34 Issue 8 Pages 1273-1278
    Published: August 01, 2011
    Released: August 01, 2011
    JOURNALS FREE ACCESS
    Therapeutic monoclonal antibodies (MAbs) with high specificity and fewer adverse effects are becoming widely used for the treatment of various diseases. MAbs need to be stored and administered at high concentrations in solution, the conditions under which MAbs may aggregate. As aggregated MAbs compromise their safety and efficacy, aggregation should be prevented; thus, it is important to analyze the aggregation states of MAbs in detail. We obtained 2 MAbs against dinitrophenol (DNP) that exhibited different aggregation properties: anti-DNP1 exhibited a much higher aggregate content (dimer or trimer) than anti-DNP2 when analyzed by size-exclusion chromatography (SEC). As anti-DNP1 had a longer complementarity-determining region 3 (CDR3) light chain than anti-DNP2 by 2 amino acid residues, we hypothesized that the increased aggregation of DNP1 was due to these extra residues; therefore, we prepared mutant antibodies with shorter CDR3s to compare their aggregation properties. Anti-DNP1-ΔEI, with the same CDR3 length as anti-DNP2, exhibited no aggregates as expected. Anti-DNP1-ΔI, with 1 additional residue, exhibited a smaller peak than wild-type (WT) in SEC, whereas this mutant exhibited stronger thioflavin T fluorescence than WT, which is indicative of amyloid formation. In addition, the anti-DNP1-ΔI solution (but not others) became opalescent at 4°C and exhibited large visible particles, which are undetectable by SEC. The fragment antigen-binding region of this mutant was found to have lower thermal stability than the others by differential scanning calorimetry. These data suggest that diverse analytical methods should be applied to evaluate MAb aggregation, in addition to the commonly used SEC.
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  • Chun Ying Xie, Wei Yang, Jun Ying, Qing Chun Ni, Xue Diao Pan, Jin Hua ...
    Type: Regular Article
    2011 Volume 34 Issue 8 Pages 1279-1286
    Published: August 01, 2011
    Released: August 01, 2011
    JOURNALS FREE ACCESS
    δ-Elemene, an antitumor component, is a chemical compound isolated from Curcuma wenyujin, a Chinese traditional herb. We examined whether δ-elemene could affect apoptosis in human lung carcinoma mucoepidermoid NCI-H292 cells, and test whether and how the over-expression of B-cell lymphoma-2 (Bcl-2) and B-cell lymphoma extra large (Bcl-xL) could off-set the effect of δ-elemene on cell growth. The result demonstrated that δ-elemene significantly induced apoptosis of NCI-H292, as shown by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, DNA fragmentation measurement, Annexin V (AnV) binding of externalized phosphatidylserine and the mitochondrial probe JC-1 using flow cytometry. Treatment of NCI-H292 with δ-elemene increased both p38 mitogen-activated protein kinase (MAPK) and inducible nitric oxide synthese (iNOS) levels, suggesting these two molecules maybe relate to the apoptotic effect of δ-elemene. The cells with Bcl-2 or Bcl-xL over-expression showed an elevation of nuclear factor kappa B (NF-kappa B) activity, accompanying a significant reduction of δ-elemene-induced apoptosis. Furthermore, inhibition of NF-kappa B by IkBαSR, which is a powerful inhibitor of NF-kappa B, restored the ability of δ-elemene to induce apoptosis in the cells transfected with Bcl-2. These data strongly indicated that the apoptotic effect of δ-elemene on NCI-H292 was closely associated with the activity of NF-kappa B, which was up-regulated by Bcl-2 and Bcl-xL. In conclusion, δ-elemene induced apoptosis in NCI-H292 cells. The apoptotic effect of δ-elemene could be significantly offset by over-expression of either Bcl-2 or Bcl-xL. Bcl-2 and Bcl-xL were able to increase the activity of NF-kappa B, which was a known anti-apoptotic molecule in human lung cancer cells.
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  • Patamaporn Pruksakorn, Masayoshi Arai, Liu Liu, Prashini Moodley, Will ...
    Type: Regular Article
    2011 Volume 34 Issue 8 Pages 1287-1290
    Published: August 01, 2011
    Released: August 01, 2011
    JOURNALS FREE ACCESS
    In the course of our search for anti-dormant mycobacterial substances from marine organisms, we previously isolated three new aminolipopeptides, named trichoderins A, A1 and B, from the culture of the marine sponge-derived fungus of Trichoderma sp. and determined their chemical structures. To identify the gene that could confer a resistance to trichoderin A, we prepared transformants of Mycobacterium (M.) smegmatis, which were transformed with the genomic DNA library of M. bovis BCG constructed in the multi-copy shuttle cosmid pYUB145. Then, the transformant of M. smegmatis, which over-expressed a part of genes that coded mycobacterial ATP synthase, was found to exhibit a resistance to trichoderin A. In addition, trichoderin A reduced ATP contents in M. bovis BCG. These findings elucidated that the anti-mycobacterial activity of trichoderins comes from the inhibition of ATP synthesis.
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  • Shen Li, Xiao-Ping Pu
    Type: Regular Article
    2011 Volume 34 Issue 8 Pages 1291-1296
    Published: August 01, 2011
    Released: August 01, 2011
    JOURNALS FREE ACCESS
    In the present study, we investigated the neuroprotective effects of kaempferol in the mouse model of Parkinson's disease, which was induced by neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). We confirmed that MPTP led to behavioral deficits, depletion of dopamine and its metabolites, reduction in superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX) activity, and the elevation of malondialdehyde (MDA) levels in the substantia nigra. When administered prior to MPTP, kaempferol improved motor coordination, raised striatal dopamine and its metabolite levels, increased SOD and GSH-PX activity, and reduced the content of MDA compared with mice treated with MPTP alone. Immunohistochemical studies using anti-tyrosine hydroxylase (TH) antibody showed that medication of kaempferol could prevent the loss of TH-positive neurons induced by MPTP. Taken together, we propose that kaempferol has shown anti-parkinsonian properties in our studies. More work is needed to explore detailed mechanisms of action.
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  • Tetsuo Adachi, Kazunari Aida, Hiroko Nishihara, Tetsuro Kamiya, Hiroka ...
    Type: Regular Article
    2011 Volume 34 Issue 8 Pages 1297-1300
    Published: August 01, 2011
    Released: August 01, 2011
    JOURNALS FREE ACCESS
    The initial clinical stage of diabetic retinopathy (DR) is characterized by the development of intraretinal microvascular abnormalities. The increased formation of reactive oxygen species (ROS) is thought to be a key event in the pathogenesis of DR. Extracellular-superoxide dismutase (EC-SOD) is an anti-inflammatory enzyme that is distributed mainly in vascular cells and protects cells from ROS by scavenging superoxide anion. Treatment with cobalt chloride (CoCl2) decreased the expression of EC-SOD but not other SOD isozymes in pericytes accompanied with an increase of intracellular ROS production. Pre-treatment with N-acetylcysteine (NAC) significantly suppressed the ROS production and down-regulation of EC-SOD. We observed the activation of caspase-3 and DNA fragmentation as signs of apoptotic process by CoCl2 treatment. In addition, these phenomena were significantly inhibited by pre-treatment with NAC. EC-SOD enhancer 4-phenyl butyric acid also suppressed the caspase-3 activation. It is known that the presence of a high level of EC-SOD throughout the vessel walls might have an important protective role against superoxide in the vascular system. The decrease in EC-SOD expression accompanied with elevation of ROS level in pericytes under hypoxia might induce and/or promote the ROS-triggered apoptosis of pericytes and the development of pathogenesis in DR.
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  • Martha Ines Burgos, Ricardo Ariel Fernández, María Soled ...
    Type: Regular Article
    2011 Volume 34 Issue 8 Pages 1301-1306
    Published: August 01, 2011
    Released: August 01, 2011
    JOURNALS FREE ACCESS
    Tetracycline (TC) derivatives are extensively used as antibiotics in human and animal medicine and, very recently, they have been screened as anti-amyloidogenic drugs. Anhydrotetracycline (AHTC) is one of the major degradation products of TC that has been linked to several side effects of the drug. We evaluated the interaction of AHTC with bovine serum albumin (BSA), one of the main carriers of amphiphilic molecules in blood, using three complementary analytical methods: fluorescence spectroscopy, isothermal titration calorimetry and differential scanning calorimetry. AHTC bound to BSA with an association constant in the order of 105 M−1. Drug binding was enthalpically and entropically driven and seemed to involve hydrophobic interactions. AHTC fluorescence enhancement and hypsochromic shifts observed upon binding suggested a low-polarity location excluded from water for the bound drug. Our data are useful for evaluating the biodisponibility of the pharmacophore and the dynamic distribution of the toxic derivative.
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  • Takaharu Ishibashi, Tomoko Miwa, Naoki Nishizawa, Ikumi Shinkawa, Junk ...
    Type: Regular Article
    2011 Volume 34 Issue 8 Pages 1307-1313
    Published: August 01, 2011
    Released: August 01, 2011
    JOURNALS FREE ACCESS
    The regulatory role of plasma nitrosothiols (R-SNOs) under steady-state conditions and their possible contribution to pharmacological vasodilation were systematically examined in anesthetized rabbits. Nitrosocystein (Cys-NO), S-nitrosoglutathione (G-SNO), and S-nitrosoalbumin (Alb-SNO) were determined by HPLC-Saville's method with respective sensitivities of 1, 1, and 5 nM. These R-SNOs were not detected under steady-state conditions even in the presence of N-ethylmaleimide, a thiol protective agent used to prevent transnitrosation of R-SNOs. Development of plasma Alb-SNO below 300 nM was observed after intravenous injection (i.v.) of nitric oxide (NO) solution (0.1 to 3 ml/g), NOC7 (an NO releasing agent, above 1 μg/kg), and a low dose of Alb-SNO (10 nmol/kg). However, blood pressure was not significantly reduced by NO solution or Alb-SNO. Intravenous injection of a high dose of Alb-SNO (300 nmol/kg) significantly reduced blood pressure with the appearance not only Alb-SNO in micromolar level in plasma, but also G-SNO in lesser degree. Conversely, the hypotensive effect of Cys-NO (300 nmol/kg, i.v.) and G-SNO (300 nmol/kg, i.v.) accompanied development of Alb-SNO (micromolar level), but not Cys-NO or G-SNO in plasma. R-SNOs were not found in plasma during profound hypotension induced by acetylcholine (10 and 30 μg/kg/min, continuous i.v.), glyceryl trinitrate (100 μg/kg, i.v.), sodium nitroprusside (100 μg/kg, i.v.), and isosorbide dinitrate (300 μg/kg, i.v.). These results indicate that R-SNOs do not play an important role under unstimulated condition. In addition, plasma R-SNOs may not be involved in pharmacological vasodilation where contributions of NO or R-SNOs are suggested.
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  • Makoto Kanamori, Masayuki Seki, Akari Yoshimura, Toshiki Tsurimoto, Sh ...
    Type: Regular Article
    2011 Volume 34 Issue 8 Pages 1314-1318
    Published: August 01, 2011
    Released: August 01, 2011
    JOURNALS FREE ACCESS
    Werner interacting protein 1 (WRNIP1) that is highly conserved from Escherichia coli to human was originally identified as a protein that interacts with the Werner syndrome responsible gene product (WRN). Here, human WRNIP1 and WRN are shown to bind to template-primer DNA, and WRNIP1, but not WRN, requires ATP for DNA binding. Under conditions of a limiting amount of WRN, WRNIP1 facilitated binding of WRN to DNA in a dose-dependent manner. However, WRNIP1 did not stimulate the DNA helicase activity of WRN, and WRN displaced pre-bound WRNIP1 from DNA. Functional relationships between WRNIP1 and WRN will be discussed.
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  • Tingting Lu, Yongjun Jiang, Zhiming Zhou, Xuanye Yue, Ning Wei, Zhaoya ...
    Type: Regular Article
    2011 Volume 34 Issue 8 Pages 1319-1324
    Published: August 01, 2011
    Released: August 01, 2011
    JOURNALS FREE ACCESS
    Ginsenoside Rb1 (GRb1) has been shown to benefit many central nervous system (CNS) disorders, including stroke. However, its bioavailability is low after oral administration due to poor absorption. Intranasal administration has been considered as an effective method for central nervous system drug delivery for its brain-targeting effect. Here, whether intranasal GRb1 could ameliorate cerebral ischemia/reperfusion injury was investigated. First, the concentration of GRb1 in brain tissues and plasma after intranasal and intravenous delivery was calculated using HPLC-MS/MS methods in male Sprague-Dawley rats (250±10 g). Intranasal GRb1 was considered brain-targeting if the value of the drug targeting index (DTI) was greater than 1. Rats were subjected to 1.5 h middle cerebral artery occlusion (MCAO) and were killed 24 h after reperfusion. The neuroprotective effects were measured using 2,3,5-triphenyltetrazolium chloride (TTC) staining and Nissl staining. Immunoblotting of LC3 and Beclin 1, crucial autophagy-related proteins, was used to monitor the state of autophagy. With a local bioavailability of 10.28—32.48% and DTI of 7.35—23.22 in different brain regions, intranasal GRb1 was determined to be brain-targeting. Less infarct volume and more intact neuronal structure were observed in the GRb1 group. GRb1 also restored the elevation of LC3 and Beclin 1. Our work suggests that intranasal GRb1 exerts brain-targeting effects and that a single dose of intranasal GRb1 immediately after MCAO ameliorates ischemia/reperfusion insult. Autophagy is involved in these beneficial effects.
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  • Shigeharu Oie, Akiko Obayashi, Hirofumi Yamasaki, Hiroyuki Furukawa, T ...
    Type: Regular Article
    2011 Volume 34 Issue 8 Pages 1325-1329
    Published: August 01, 2011
    Released: August 01, 2011
    JOURNALS FREE ACCESS
    To evaluate disinfection methods for environments contaminated with bioterrorism-associated microorganism (Bacillus anthracis), we performed the following experiments. First, the sporicidal effects of sodium hypochlorite on spores of five bacterial species were evaluated. Bacillus atrophaeus was the most resistant to hypochlorite, followed in order by B. anthracis, Clostridium botulinum and Clostridium tetani, and Clostridium difficile. Subsequently, using B. atrophaeus spores that were the most resistant to hypochlorite, the sporicidal effects of hypochlorite at lower pH by adding vinegar were evaluated. Hypochlorite containing vinegar had far more marked sporicidal effects than hypochlorite alone. Cleaning with 0.5% (5000 ppm) hypochlorite containing vinegar inactivated B. atrophaeus spores attached to vinyl chloride and plywood plates within 15 s, while that not containing vinegar did not inactivate spores attached to cement or plywood plates even after 1 h. Therefore, the surfaces of cement or plywood plates were covered with gauze soaked in 0.5% hypochlorite containing vinegar, and the sporicidal effects were evaluated. B. atrophaeus spores attached to plywood plates were not inactivated even after 6 h, but those attached to cement plates were inactivated within 5 min. On the other hand, covering the surfaces of plywood plates with gauze soaked in 0.3% peracetic acid and gauze soaked in 2% glutaral inactivated B. atrophaeus spores within 5 min and 6 h, respectively. These results suggest that hypochlorite containing vinegar is effective for disinfecting vinyl chloride, tile, and cement plates contaminated with B. anthracis, and peracetic acid is effective for disinfecting plywood plates contaminated with such microorganism.
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Notes
  • Hisae Oku, Yuko Ogawa, Emiko Iwaoka, Kyoko Ishiguro
    Type: Note
    2011 Volume 34 Issue 8 Pages 1330-1333
    Published: August 01, 2011
    Released: August 01, 2011
    JOURNALS FREE ACCESS
    Allergy-preventive activity of flower buds of Lonicera japonica THUNB. was found in the 35% EtOH extract (LJ) using an in vivo assay, The assay system uses monitoring of a decrease in blood flow (BF) in the tail vein of mice subjected to sensitization with hen-egg white lysozyme (HEL). Bioassay-guided fractionation of the 35% EtOH extract led to isolation of chlorogenic acid (1) and three known iridoid derivatives, loganin (2), secoxyloganin (3) and sweroside (4), all of which inhibited the BF decrease. This suggested that the flower buds of L. japonica and compounds isolated from them have allergy-preventive properties. The structure–activity relationship of iridoid derivatives, morroniside (5), geniposide (6), asperuloside (7), aucubin (8) and catalpol (9), were also tested using the same bioassay method. Compounds 25 and 9 having the sp3 atom at C-8 showed an allergy-preventive effect, while compounds 6, 7 and 8 having a double bond at C-7, C-8 did not.
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  • Mareshige Kojoma, Shigeki Hayashi, Toshiro Shibata, Yutaka Yamamoto, H ...
    Type: Note
    2011 Volume 34 Issue 8 Pages 1334-1337
    Published: August 01, 2011
    Released: August 01, 2011
    JOURNALS FREE ACCESS
    Cultivated licorice plants (Glycyrrhiza uralensis FISCH.) contain smaller amounts of the triterpene saponin glycyrrhizin than wild licorice plants. To resolve this problem and to breed strains with high-glycyrrhizin content we determined the glycyrrhizin content of 100 samples of G. uralensis that were propagated from seed and grown under the same conditions in the field for 5 years. There was a 10.2-fold variation in glycyrrhizin content among these plants, ranging from 0.46 to 4.67% (average 2.11±0.90%). There was also a wide variation in liquiritin content, ranging from 0.11 to 2.65% (average 1.00±0.49%). The glycyrrhizin content was positively correlated with that of liquiritin in the taproots (r2=0.5525). Our results indicate that there are various genetic strains for glycyrrhizin and liquiritin synthesis within a population of plants propagated from seed. The selected high-glycyrrhizin and liquiritin strains will be useful for licorice production and studies on biosynthetic analysis of glycyrrhizin and liquiritin.
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  • Satoru Sonoke, Toshihiro Ueda, Kae Fujiwara, Kenji Kuwabara, Junichi Y ...
    Type: Note
    2011 Volume 34 Issue 8 Pages 1338-1342
    Published: August 01, 2011
    Released: August 01, 2011
    JOURNALS FREE ACCESS
    We have developed a galactose-modified cationic liposome for delivery of small interfering RNA (siRNA) to the liver. The liposomes were designed to be transported into hepatocytes via the asialoglycoprotein receptor, which recognizes galactose residues. The liposomes contained a novel galactose-modified lipid, 1,2-dioleoyl-sn-glycerol-3-phosphatidyl-N-(1-deoxylactito-1-yl)ethanolamine (GDOPE). Delivery of siRNA to hepatocytes by the liposomes was evaluated by measuring the gene-silencing activity of liposome : siRNA complexes in two human hepatoma cell lines. A formulation with a cationic lipid : GDOPE ratio of 3 : 5 by weight, LIC-G5, showed the strongest activity. In mice, intravenous injection of LIC-G5 complexed with 3H-labeled siRNA led to accumulation of radioactivity in the liver. When the hepatic cellular uptake was determined after intravenous injection into mice followed by collagenase liver perfusion, the distribution of siRNA to parenchymal cells was 1.9 times higher when LIC-G5 rather than nongalactosylated LIC was used as the carrier. The concentration of siRNA accumulated was 45 μg/ml, 30 times the concentration that produced strong gene silencing in vitro and therefore presumably sufficient for a therapeutic effect. Because increasing the cationic-lipid content of a liposome carrier generally enhances the uptake of siRNA by the liver at the expense of increased cell toxicity, we used only a moderate amount of cationic lipid in our galactose-modified carrier. LIC-G5 enhanced the uptake of siRNA by the liver without cytotoxic effects and is a promising candidate delivery system for liver-targeted siRNA therapy.
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  • Tomomi Osaki, Yoshimi Uchida, Jun Hirayama, Hiroshi Nishina
    Type: Note
    2011 Volume 34 Issue 8 Pages 1343-1347
    Published: August 01, 2011
    Released: August 01, 2011
    JOURNALS FREE ACCESS
    In most species, solar light is both a DNA-damaging agent and the key entraining stimulus for the endogenous circadian clock. The zebrafish is an attractive vertebrate system in which to study the influence of light on gene expression because the DNA repair proteins and circadian oscillators in this species are light-responsive. At the molecular level, light treatment of zebrafish cells induces the production of reactive oxygen species (ROS). ROS both alters the reduction–oxidation (redox) state of these cells and stimulates intracellular extracellular signal-regulated kinase (ERK)/mitogen activated protein kinase (MAPK) cascades that transduce photic signals activating the transcription of particular light-responsive genes, including some clock genes and some DNA repair genes involved in photoreactivation. To date, however, the phototransducing molecules responsible for light-dependent ROS production have not been identified. Flavin-containing oxidases, such as reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidase, are versatile flavoenzymes that catalyze molecular oxidation in numerous metabolic pathways. Importantly, light induces the photoreduction of the flavin adenine dinucleotide (FAD) moiety in these oxidases, leading to ROS production. Here, we show in cultured zebrafish cells that diphenyleneiodonium chloride (DPI), an inhibitor of NADPH oxidase, both suppresses ERK/MAPK activation and efficiently reduces light-dependent expression of clock and photoreactivation genes. Our results suggest that flavin-containing oxidases may be responsible for light-dependent ROS production and thus light-dependent gene expression in zebrafish. Our findings also support the existence of a regulatory link between photoreactivation and the circadian clock in this species.
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  • Shota Warashina, Takashi Nakamura, Hideyoshi Harashima
    Type: Note
    2011 Volume 34 Issue 8 Pages 1348-1351
    Published: August 01, 2011
    Released: August 01, 2011
    JOURNALS FREE ACCESS
    In a previous report, we described the development of lipid envelope-type nanoparticles (MEND) modified with octaarginine (R8) and a pH-sensitive fusogenic peptide (GALA) for delivering short interference RNA (siRNA) to mouse dendritic cells (DCs). A20 was recently reported to be a negative regulator of the toll-like receptor and the tumor necrosis factor receptor signaling pathways. Although A20 would be expected to be a useful target for boosting the effects of adjuvants in DC immunotherapy, limited information is available regarding the use of A20-silenced DC by an original non-viral vector. In this study, we loaded anti-A20 siRNA into a MEND and investigated the gene knockdown activity in DC and the immunological functions of A20-silenced DC. The use of a MEND resulted in a significant A20 knockdown effect, and the A20-silenced DC resulted in an enhanced production of proinflammatory molecules, after lipopolysaccharide (LPS) stimulation. The expression of co-stimulatory molecules by LPS stimulation was also increased in the A20-silenced DC. The findings reported herein show that a MEND loaded with anti-A20 siRNA is a potent non-viral vector that has the ability to enhance the adjuvant effect of LPS in DC.
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  • Noriyuki Iwabuchi, Jin-Zhong Xiao, Tomoko Yaeshima, Keiji Iwatsuki
    Type: Note
    2011 Volume 34 Issue 8 Pages 1352-1355
    Published: August 01, 2011
    Released: August 01, 2011
    JOURNALS FREE ACCESS
    We investigated whether the oral administration of Bifidobacterium longum BB536 could ameliorate influenza virus (IFV) infection in a mice model. Mice were orally administrated BB536 or saline for 2 weeks and then infected with IFV. Orally administered BB536 significantly alleviated symptoms, reduced the loss of body weight, and inhibited viral proliferation in the lungs relative to the control group findings. Histopathological findings in the lungs were improved in the BB536 group compared to control group findings. There was no significant difference in the levels of interleukin-6 (IL-6), interferon-γ (IFN-γ), IL-10 and IL-12p40 in the lungs between the groups, but the levels of IL-6 and IFN-γ were lower (p=0.076, 0.103, respectively) in the BB536 group compared with those of control group. The levels of IL-6 and IL-10 correlated significantly with the values of weight loss, and the levels of IFN-γ correlated with the virus titers in the lungs. These results suggested the potential of the oral administration of BB536 in ameliorating IFV infection and the possible involvement of anti-inflammatory effects of BB536 in the anti-infection effects against IFV.
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