The 5′-AT-rich half-site of Maf recognition element: a functional target for bZIP transcription factor Maf

The Maf family of proteins are a subgroup of basic region-leucine zipper (bZIP) transcription factors, which recognize a long palindromic DNA sequence [TGCTGAC(G)TCAGCA] known as the Maf recognition element (MARE). Interestingly, the functional target enhancer sequences present in the αA-crystallin gene contain a well-conserved half-site of MARE rather than the entire palindromic sequence. To resolve how Maf proteins bind to target sequences containing only MARE half-sites, we examined their binding activities using electrophoretic gel mobility shift assays as well as in vitro and in vivo reporter assays. Our results indicate that the 5′-flanking region of the MARE half-site is required for Maf proteins to bind both in vitro and in vivo. The critical 5′-flanking sequences for c-Maf were determined by a selection and amplification binding assay and show a preference for AT-rich nucleotides. Furthermore, sequence analysis of the regulatory regions of several target genes also suggests that AT-rich sequences are important. We conclude that Maf can bind to at least two types of target sequences, the classical MARE (palindrome type) and a 5′-AT-rich MARE half-site (half-site type). Our results provide important new insights into the DNA binding and site selection by bZIP transcription factors

New immunization procedure for production of monoclonal antibodies which recognize carbohydrate of glycoprotein

糖タンパク質糖鎖に対するモノクローナル抗体を効率よくとるための免疫方法の検討をおこなった。抗原としては糖鎖のがん性変化のひとつであるbisecting　N－acetyiglucosamine構造をもつオボムコイド（OVM）をとりあげた。OVM全分子を通常のフロインド完全アジュバントでくりかえし感作する方法では糖鎖を認識するモノクローナル抗体をとることはできなかった。またOVMをリポソームニ重膜に挿入する方法でも同様であった。これに対してOVMからプロナーゼ消化により糖ペプチドを調製し，アジュバント活性をもつリピドAを共存させたリポソームに共有結合した抗原を感作したマウスからは高率に糖鎖と反応するモノクローナル抗体をとることができた。しかしながらこれらの抗体はいずれもサブタイプはIgMであった。New immunization method for production of monoclonal anitibodies which recognize oligosaccharide portion of glycoprotein was developed. Conventional immunization method in which glycoprotein was emulsified with Freund's complete adjuvant could not produce anti-carbohydrate monoclonal antibody. Glycopeptide which was prepared by pronase digestion of glycoprotein conjugated liposomes containing lipid A of Salmonella minnesota were revealed good antigen for prodution of anti-carbohydrate monoclonal anitibodies. By this new immunization method several monoclonal antibodies which recognize mainly carbohydrate portion of ovomucoide were established

Coronary angioplasty ameliorates hypoperfusion-induced endothelial dysfunction in patients with stable angina pectoris

Objectives.This study sought to investigate the effect of coronary angioplasty on chronic hypoperfusion-induced endothelial dysfunction in patients with coronary heart disease.Background.The endothelium is an important component for organ flow regulation. Ischemia with or without reperfusion is known to cause endothelial dysfunction. We tested the hypothesis that chronic hypoperfusion impairs endothelial function in the angiographically normal coronary artery segment distal to stenosis and that the impairment by chronic hypoperfusion is reduced by coronary angioplasty.Methods.In 13 patients with stable angina pectoris, substance P (10, 30 and 100 pmol) and nitroglycerin (200 μg) were sequentially infused into the coronary artery in a cumulative manner on the day after coronary angioplasty. In 10 of these patients, vascular responses to these agents were again investigated 3 months after angioplasty. Changes in vascular diameter were evaluated in vessels located proximal and distal to the target lesion, both of which were angiographically normal, by performing computer-assisted quantitative coronary angiography. In five patients, the transstenotic pressure gradient was also measured with a pressure sensor-mounted guide wire before angioplasty.Results.On the day after angioplasty, the magnitude of dilation by substance P in distal segments was significantly less than that in proximal segments and inversely correlated with the transstenotic pressure gradient (p < 0.05) and lesion stenosis (p < 0.05). There was no difference in nitroglycerin-induced vasodilation between the two vessel segment groups. Three months later, the impaired response to substance P in the distal segment was restored to normal.Conclusions.We conclude that chronic hypoperfusion impairs endothelium-dependent dilation of coronary artery distal to critical stenosis in patients with ischemic heart disease and that coronary angioplasty ameliorates the endothelial dysfunction within 3 months

Expression and functional analysis of Nr2e3, a photoreceptor-specific nuclear receptor, suggest common mechanisms in retinal development between avians and mammals

The photoreceptor-specific nuclear receptor (PNR; Nr2e3) is a transcription factor important for retinal development. We report here the identification and expression analysis of the avian Nr2e3. Nr2e3 mRNA is expressed in the photoreceptor layer of the neural retina during early stages of chick embryogenesis. Its temporal expression is distinct from that of a related nuclear receptor, Tlx. Chick Nr2e3 recognizes and binds to the same target DNA sequence as its vertebrate orthologs. Functional assays revealed that chick Nr2e3 acts as a transcriptional repressor. Our results suggest that Nr2e3 plays a common role in retinal development in vertebrates

Current Performance and On-Going Improvements of the 8.2 m Subaru Telescope

An overview of the current status of the 8.2 m Subaru Telescope constructed and operated at Mauna Kea, Hawaii, by the National Astronomical Observatory of Japan is presented. The basic design concept and the verified performance of the telescope system are described. Also given are the status of the instrument package offered to the astronomical community, the status of operation, and some of the future plans. The status of the telescope reported in a number of SPIE papers as of the summer of 2002 are incorporated with some updates included as of 2004 February. However, readers are encouraged to check the most updated status of the telescope through the home page, http://subarutelescope.org/index.html, and/or the direct contact with the observatory staff.Comment: 18 pages (17 pages in published version), 29 figures (GIF format), This is the version before the galley proo

青年期女子の愛着スタイルと情緒的応答性の関連

The purpose of this study was to investigate the relationships between attachment styles of female university students and their emotional availability to infants. Attachment styles were assessed by two questionnaires of attachment internal working models (IWM and ECR) and emotional availability was assessed by Japanese edition of IFEEL Pictures (pictures of infants\u27 facial expressions). Range of used categories was correlated positively and number of popular responses was correlated negatively to secure scores. Anxious individuals showed more popular responses to infant facial expressions. These results suggested that young female adults with secure attachment have the readiness to read emotions and motivations of infants more flexibly