Impact of polyplex micelles installed with cyclic RGD peptide as ligand on gene delivery to vascular lesions

Gene therapy is expected to open a new strategy for the treatment of refractory vascular diseases, so the development of appropriate gene vectors for vascular lesions is needed. To realize this requirement with a non-viral approach, cyclo(RGDfK) peptide (cRGD) was introduced to block copolymer, poly(ethylene glycol)-block-polycation carrying ethylenediamine units (PEG-PAsp(DET)). cRGD recognizes αvβ3 and αvβ5 integrins, which are abundantly expressed in vascular lesions. cRGD-conjugated PEG-PAsp(DET) (cRGD-PEG-PAsp(DET)) formed polyplex micelles through complexation with plasmid DNA (pDNA), and the cRGD-PEG-PAsp(DET) micelles achieved significantly more efficient gene expression and cellular uptake as compared with PEG-PAsp(DET) micelles in endothelial cells and vascular smooth muscle cells. Intracellular tracking of pDNA showed that cRGD-PEG-PAsp(DET) micelles were internalized via caveolae-mediated endocytosis, which is associated with a pathway avoiding lysosomal degradation, and that PEG-PAsp(DET) micelles were transported to acidic endosomes and lysosomes via clathrin-mediated endocytosis. Further, in vivo evaluation in rat carotid artery with a neointimal lesion revealed that cRGD-PEG-PAsp(DET) micelles realized sustained gene expression, while PEG-PAsp(DET) micelles facilitated rapid but transient gene expression. These findings suggest that introduction of cRGD to polyplex micelles might create novel and useful functions for gene transfer and contribute to the establishment of efficient gene therapy for vascular diseases

Recognition of 2′-hydroxyl groups by Escherichia coli ribonuclease HI

AbstractIn order to investigate the hydrogen-bonding interactions between Escherichia coli ribonuclease HI and the 2′-hydroxyl functions of the substrate, oligonucleotide duplexes containing 2′-amino-2′-deoxyuridine or 2′-fluoro-2′-deoxyuridine at a specific site were used, and their affinities for the enzyme were determined by kinetic analyses. The results indicate that the hydroxyl groups of the nucleoside 3′-adjacent to the cleaved phosphodiester linkage and the second nucleoside 5′ to the cleaved phosphodiester act as both a proton donor and an acceptor and as a proton acceptor, respectively, in the enzyme-substrate complex. A molecular model was constructed using the interactions derived from the results

Nasal double DNA adjuvant induces salivary FimA-specific secretory IgA antibodies in young and aging mice and blocks Porphyromonas gingivalis binding to a salivary protein

Background: We previously showed that nasal administration of a combination of dendritic cell (DC) targeted DNA plasmid expressing Flt3 ligand and CpG oligodeoxynucleotides 1826 as a mucosal adjuvant (double adjuvant, DA) provoked protective immunity in the upper respiratory tract of young adult and aging mice. Here, we investigated whether the nasal DA system induces secretory (S)IgA antibodies (Abs) toward recombinant fimbrillin (rFimA) of Porphyromonas gingivalis (P. gingivalis) in the saliva of young adult and aging mice. Further, we examined the functional applicability of rFimA-specific salivary SIgA Abs. Methods: BALB/c mice (8- or 48-week-old) were nasally immunized with rFimA plus DA three times at weekly intervals. Control mice were nasally administered rFimA alone. Saliva samples were collected 1 week after the final immunization, and were subjected to rFimA-specific ELISA. To examine the functional applicability of rFimA-specific SIgA Abs, IgA-enriched saliva samples were subjected to an inhibition assay in order to assess the numbers of P. gingivalis cells bound to the salivary protein statherin. Results: The 8- and 48-week-old mice administered nasal rFimA plus DA showed significantly increased levels of rFimA-specific SIgA Abs in saliva and elevated numbers of CD11c+ DCs in sublingual glands (SLGs), periglandular lymph nodes (PGLNs) and submandibular glands (SMGs) as well as nasopharyngeal-associated lymphoid tissues (NALT) compared to mice administered rFimA alone. Further, rFimA-specific SIgA Abs-containing saliva, in which IgG Abs of 8- and 48-week-old mice administered nasal rFimA plus DA were removed, significantly inhibited binding of P. gingivalis to the salivary protein. Conclusions: These findings show that this DA system could be an effective nasal vaccine strategy for the enhancement of P. gingivalis-specific protective immunity in the oral cavity of adolescents and older individuals

気管支喘息患者における白血球ロイコトリエン産生能に対する不飽和脂肪酸食の効果に影響する因子

Dietary supplementation with perilla seed oil, a vegetable oil rich in α -lin- olenic acid, inhibits the generation of leukotrienes(LTs) by leucocytes in patients with bronchial asthma. We examined the factors that affect the suppression of LT generation by leucocytes with perilla seed oil-rich supplementation in patients with asthma, by comparing the clinical features of patients with asthma, whose generation of leukotriene (LT) C4 was suppressed by dietary supplementation with perilla seed oil (n-3 fatty acids) (group A), with those of patients who showed no suppression of LTC4 generation (group B). Group A showed a significant increase in the generation of LTB4 and L TC4 by leucocytes after corn oil-rich supplementation (n-6 fatty acids), and a significant decrease in the generation of LTB4 and LTC4 after perilla seed oil-rich supplementation (n-3 fatty acid). However, this was not observed in group B. The level of serum IgE and peak expiratory flow (PEF) in group A were significantly higher than in group B. Furthermore, the serum levels of LDL-cholesterol, β-lipoprotein and　phospholipid were significantly lower in group A than in group B. These results suggest that the clinical features differ between these two asthmatic populations with respect to suppression of LTB4 and LTC4 generation by n-3 fatty acids in perilla seed oil-rich supplementation.a-リノレン酸の豊富なエゴマ油の食事は気管支喘息患者の白血球ロイコトリエン(LT)産生能を抑制する｡気管支喘息患者の内,エゴマ油食によりLTC4の産生が抑制された群(A群)と抑制されない群(B群)の臨床データを比較することにより,気管支喘息患者の白血球ロイコトリエン産生能に影響する因子を検討した｡A群はコーン油(n-6系脂肪酸)の豊富な食事後,白血LTB4,LTC4の産生能が増加し,エゴマ油(n-3系脂肪酸)の豊富な食事後LTB4,LTC4の産生能が減少した｡これらの変化はB群では認められなかった｡A群のIgE値,ピークフロー(PEF)値はB群に比し,有意に高値であった｡またLDL-コレステロール,β-リポ蛋白,リン脂質はA群ではB群に比し,有意に低値であった｡これらの結果はエゴマ油の豊富な食事のn-3系脂肪酸によるLTB4,LTC4の産生能の抑制に関して2群の気管支喘息患者群間に臨床データの相違があることを示唆している

VLBI Monitoring of 3C 84 (NGC 1275) in Early Phase of the 2005 Outburst

Multi-epoch Very Long Baseline Interferometry (VLBI) study of the sub-pc scale jet of 3C 84 is presented. We carried out 14-epoch VLBI observations during 2006-2009 with the Japanese VLBI Network (JVN) and the VLBI Exploration of Radio Astrometry (VERA), immediately following the radio outburst that began in 2005. We confirmed that the outburst was associated with the central ~1 pc core, accompanying the emergence of a new component. This is striking evidence of the recurrence of jet activity. The new component became brighter during 2008, in contrast to the constant gamma-ray emission that was observed with the Fermi Gamma-ray Space Telescope during the same time. We found that the projected speed of the new component is 0.23c from 2007/297 (2007 October 24) to 2009/114 (2009 April 24). The direction of movement of this component differs from that of the pre-existing component by ~40 degree. This is the first measurement of kinematics of a sub-pc jet in a gamma-ray active phase. Possible detection of jet deceleration and the jet kinematics in connection with the gamma-ray emission is discussed.Comment: 5 pages, 3 figures. Accepted for publication in PAS

Involvement of oxidative stress and mucosal addressin cell adhesion molecule-1 (MAdCAM-1) in inflammatory bowel disease

The pathophysiology of inflammatory bowel disease involves excessive immune effects of inflammatory cells against gut microbes. In genetically predisposed individuals, these effects are considered to contribute to the initiation and perpetuation of mucosal injury. Oxidative stress is a fundamental tissue-destructive mechanisms that can occur due to the reactive oxygen species and reactive nitrogen metabolites which are released in abundance from numerous inflammatory cells that have extravasated from lymphatics and blood vessels to the lamina propria. This extravasation is mediated by interactions between adhesion molecules including mucosal addressin cell adhesion molecule-1 and vascular cell adhesion molecule-1 on the surface of lymphocytes or neutrophils and their ligands on endothelial cells. Thus, reactive oxygen species and adhesion molecules play an important role in the development of inflammatory bowel disease. The present review focuses on the involvement of oxidative stress and adhesion molecules, in particular mucosal addressin cell adhesion molecule-1, in inflammatory bowel disease

Endoscopic Resection of Zenker's Diverticulum

We report an endoscopically assisted total diverticulectomy for Zenker's diverticulum. Skin incisions were made at the anterior axillary line, the center of the sternum, and the neck as portals for endoscopical instruments. The skin was retracted with hooks which provided an excellent view of the working space. The diverticulum was fully exposed and resected by using a multifire endoscopic stapler. This approach is minimally invasive in comparison with the conventional open cervical approach

Changes in 12-Year First-Line Eradication Rate of Helicobacter pylori Based on Triple Therapy with Proton Pump Inhibitor, Amoxicillin and Clarithromycin

A triple therapy based on a proton pump inhibitor (PPI), amoxicillin (AMPC), and clarithromycin (CAM) is recommended as a first-line therapy for Helicobacter pylori (H. pylori) eradication and is widely used in Japan. However, a decline in eradication rate associated with an increase in prevalence of CAM resistance is viewed as a problem. We investigated CAM resistance and eradication rates over time retrospectively in 750 patients who had undergone the triple therapy as first-line eradication therapy at Nagoya City University Hospital from 1995 to 2008, divided into four terms (Term 1: 1997–2000, Term 2: 2001–2003, Term 3: 2004–2006, Term 4: 2007–2008). Primary resistance to CAM rose significantly over time from 8.7% to 23.5%, 26.7% and 34.5% while the eradication rate decreased significantly from 90.6% to 80.2%, 76.0% and 74.8%. Based on the PPI type, significant declines in eradication rates were observed with omeprazole or lansoprazole, but not with rabeprazole. A decrease in the H. pylori eradication rate after triple therapy using a PPI + AMPC + CAM has been acknowledged, and an increase in CAM resistance is considered to be a factor. From now on, a first-line eradication regimen that results in a higher eradication rate ought to be investigated