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Abdominal aortic aneurysm (AAA) is considered to be a potent life-threatening disorder in elderly individuals. Although many patients with a small AAA are detected during routine abdominal screening, there is no effective therapeutic option to prevent the progression or regression of AAA in the clinical setting. Recent advances in molecular biology have led to the identification of several important molecules, including microRNA and transcription factor, in the process of AAA formation. Regulation of these factors using nucleic acid drugs is expected to be a novel therapeutic option for AAA. Nucleic acid drugs can bind to target factors, mRNA, microRNA, and transcription factors in a sequence-specific fashion, resulting in a loss of function of the target molecule at the transcriptional or posttranscriptional level. Of note, inhibition of a transcription factor using a decoy strategy effectively suppresses experimental AAA formation, by regulating the expression of several genes associated with the disease progression. This review focuses on recent advances in molecular therapy of using nucleic acid drugs to treat AAA.

Transactivation of inflammatory and immune mediators in asthma is tightly regulated by nuclear factor κB (NF-κB) and signal transducer and activator of transcription 6 (STAT6). Therefore, we investigated the efficacy of simultaneous inhibition of NF-κB and STAT6 using a chimeric decoy strategy to prevent asthma exacerbation. The effects of decoy oligodeoxynucleotides were evaluated using an ovalbumin-induced mouse asthma model. Ovalbumin-sensitized mice received intratracheal administration of decoy oligodeoxynucleotides 3 days before ovalbumin challenge. Fluorescent-dye-labeled decoy oligodeoxynucleotides could be detected in lymphocytes and macrophages in the lung, and activation of NF-κB and STAT6 was inhibited by chimeric decoy oligodeoxynucleotide transfer. Consequently, treatment with chimeric or NF-κB decoy oligodeoxynucleotides protected against methacholine-induced airway hyperresponsiveness, whereas the effect of chimeric decoy oligodeoxynucleotides was significantly greater than that of NF-κB decoy oligodeoxynucleotides. Treatment with chimeric decoy oligodeoxynucleotides suppressed airway inflammation through inhibition of overexpression of interleukin-4 (IL-4), IL-5, and IL-13 and inflammatory infiltrates. Histamine levels in the lung were reduced via suppression of mast cell accumulation. A significant reduction in mucin secretion was observed due to suppression of MUC5AC gene expression. Interestingly, the inhibitory effects on IL-5, IL-13, and histamine secretion were achieved by transfer of chimeric decoy oligodeoxynucleotides only. This novel therapeutic approach could be useful to treat patients with various types of asthma., Graphical Abstract

Angiotensin II is thought to participate in aneurysm formation, because of its ability to induce and perpetuate inflammation in the aortic wall. Because activation of renin is the first step of the renin–angiotensin system, renin inhibition could inhibit all components of this system effectively. Therefore, we examined the hypothesis that direct inhibition of renin activity could decrease the expansion of aortic aneurysm using a rabbit model. Aortic dilatation was induced by incubation with elastase around the rabbit abdominal aorta. Continuous administration of a direct renin inhibitor, aliskiren, was started at 1 week before incubation with elastase and continued for 5 weeks. Treatment with aliskiren markedly inhibited tissue renin activation and resulted in a significant reduction in angiotensin I and II production in the aneurysm wall. Consequently, the inhibition of renin activity prevented the expansion of experimental aortic aneurysm associated with preservation of the medial layer, independent of its blood pressure–lowering effect. Administration of aliskiren led to the inhibition of activation of NF-κB (nuclear factor-κB), AP-1 (activator protein-1), and CREB (cAMP response element–binding protein), which are thought to cooperatively regulate the inflammatory gene expression profile associated with aneurysm formation. As a result, treatment with aliskiren inhibited macrophage accumulation through suppression of MCP-1 (monocyte chemoattractant protein-1) and CCL4 (C-C motif chemokine ligand 4) expression, and TNF-α (tumor necrosis factor-α) production and activation of MMP-2 (matrix metalloproteinase-2) and MMP-9 (matrix metalloproteinase-9) were also suppressed in the aneurysm wall. In addition, inhibition of (pro)renin receptor elevation was also observed after treatment with aliskiren. Direct inhibition of renin activity using aliskiren prevented the progression of aortic aneurysm, suggesting it as a therapeutic option to treat abdominal aortic aneurysm.

It is well Known that the conventional method to detect presence and strength of the knocking by the ion current using the band pass filter is effective up to now. However, as compression ratio increases, there is a stationary wave in the ion current waveform and its frequency components and amplitude of the waveform are almost same as that of the knocking. Therefore the distinction of knocking and the standing wave becomes difficult by the conventional detection method. In this study, a new knocking detection technique which based on the difference of time-frequency properties of knocking and the standing wave was proposed. It first constitutes two band-pass complex real-signal mother wavelets (BC-RMW), and then thereby takes out the knocking ingredient and detects a change share of the wavelet coefficient of the knocking by using them. Furthermore, the proposed method was applied to the distinction of knocking and the standing wave and its effectiveness was inspected.

In this study, a novel direction selection method using the two-dimensional complex discrete wavelet transform (2D-CDWT) is proposed. In order to achieve arbitrary direction selection, the directional filters are first designed. Calculation procedure of directional selection can be shown as follows: (1) The 16 sub-images are generally generated from the original image by the 2D-CDWT without a down-sampling process and the 12 sub-images that correspond to the high-frequency components are selected. (2) The 12 sub-images are filtered by using the designed directional filter. (3) The down-sampling process is carried out and the resulting images are obtained. Furthermore, this method is applied to the surface analysis of a wafer, and it is confirmed that our method is effective in detecting irregular direction components.

Independent component analysis (ICA) is a useful method for blind source separation of two or more signals. We have previously proposed a new method combining ICA with the complex discrete wavelet transform (CDWT), in which voice and noise signals were separated using a new method. At that time, we used a simulated signal. In this study, we analyze measured biological signals by using a new method, and discuss its effectiveness. As an experiment, we try to separate an electromyogram (EMG) signal from an electrocardiogram (ECG) signal.

The pose estimation of 3D object of products in production line is needed beforehand. In order to perform shape measurement of objects corresponding to speed of the mass production lines before the contact measurement is done, the information of shape representation and matching of object is required. Objects are compared with its descriptor. The descriptor conceptually subtracted from each other to form a scalar metric. If the value difference is smaller, the object shape is considered closer to each other. The rotation object from its original pose can produce the information to estimate its rotation pose by using the boundary. In this paper, we proposed the method to measure 3D object pose from the designing stage of 3D CAD data.

We restore a blurred image caused by defocus of a lens using the shift-invariant wavelet transform created using RI-Spline wavelets.In a defocus blurred image,the blurring kernel varies depending on its position in the image,so a positional frequency representation,such as a wavelet space,is required in order to deblur it.To restore the defocus blur,we assume that the blurring kernel can be obtained regardless of its position in an image.In experiments we conducted using synthesized images,our method using the shift-invariant wavelet transform showed better deblurring performance than that of the ordinary wavelet transform.We also show that our method can be applied to real images if additional range data is used.

In this study, in order to improve effectiveness of the abnormal signal detection, the technique of constituting an Average Real signal Mother Wavelet (A-RMW) from two or more real signals has been proposed and its effectiveness has been confirmed. The main results obtained can be summarized as follows : (1) For constructing A-RMW by two or more signals, the Symmetric Complex Real signal Mother Wavelet (SCRMW) first was constructed respectively and then the average SCRMW was obtained by adding them. Finally the average SCRMW obtained above was normalized and the A-RMW was obtained. (2) The proposed constructing method of the SCRMW is different from the traditional Complex Real signal Mother Wavelet that the phase information of all the frequency components has been cut off and symmetric property can be obtained. (3) The proposal method is applied to the rattle noise source detection of a car, and its effectiveness is examined.

Background— Despite the advent of drug-eluting stents, restenosis after endovascular intervention is still a major limitation in the treatment of cardiovascular disease. To regulate the multiple biological mechanisms underlying restenosis, we focused on inhibition of an important transcription factor, nuclear factor-kappaB (NFκB), using a decoy strategy. Methods and Results— For site-specific application of NFκB decoy oligodeoxynucleotides into target vessels during angioplasty, we developed a balloon catheter–based delivery system combined with biocompatible nanoparticles as oligodeoxynucleotides carriers. To clarify the therapeutic effect at the site of neointima, balloon angioplasty of the rabbit carotid arteries was performed at 4 weeks after initial endothelial denudation. This delivery system exhibited successful transfer of fluorescence-labeled nanospheres into the neointima in short-term contact with target vessels, and fluorescence could be detected ≥1 week after angioplasty. Consistently, local application of NFκB decoy oligodeoxynucleotides -loaded nanospheres resulted in significant inhibition of neointimal formation, associated with inhibition of NFκB binding activity in the injured arteries. The therapeutic effects were caused by inhibition of macrophage recruitment through the suppression of monocyte chemoattractant protein-1, vascular cell adhesion molecule-1, and CC chemokine ligand 4 expression and inhibition of vascular smooth muscle cell growth via a decrease in the expression of cyclin A and proliferating cell nuclear antigen. Importantly, application of NFκB nanospheres accelerated restoration of the endothelial cell monolayer, associated with enhanced expression of phosphorylated Bcl-2 in endothelial cells. Conclusions— A drug-coated balloon catheter using NFκB decoy oligodeoxynucleotides significantly inhibited the development of neointimal hyperplasia in rabbits. The present study indicates the possibility of a novel therapeutic option to prevent restenosis after angioplasty.

Recently, much research regarding time-frequency analysis using wavelet transformation, WT, has been focused on analyzing wavelets, AW's that are derived form a mathematical approach. In the analysis in this study, the measured signal is adopted as the AW. In a general way, this method is applicable to inquiry of noise in industrial instrument and analysis of sounds, vibrations and biological signals. In an application of the proposed system, the correlation between brain waves of healthy elderly people and of senile dementia sufferers is analyzed. Brain waves are complex and non-stationary signals. To apply this method to time-varying signals such as brain waves, a new concept of instantaneous correlation factor, ICF, is introduced. This method uses WT by employing the measured signal as the AW. In this study, we use brain waves as the AW. As a conclusion, it is proved that a dominant feature of the correlation can be estimated by the ICF. Time-varying correlation is effective in the analysis of brain waves and will be useful in the diagnosis of senile dementia.

Allergic asthma is a common inflammatory disease of the airways characterized by reversible airflow obstruction and airway hyperresponsiveness (AHR) to bronchoconstrictor stimuli. The pathophysiology in asthma mainly occurs as a consequence of overexpression of the TH2 cytokines. These inflammatory factors are tightly regulated by a transcriptional network system. Nuclear factor kappaB (NFkB) and STAT6 are well known to regulate a set of gene associated with inflammatory and immune responses, and is thought to play an important role in the induction of allergic asthma. Therefore, we focused on the simultaneous inhibition of these important transcription factors using a decoy strategy to develop a novel therapeutic approach for treating asthma. For increased efficacy of decoy oligodeoxynucleotides (ODN) in vivo, we employed chimeric decoy ODN containing consensus sequences of both NFkB and STAT6 binding sites. In addition, two strands of decoy ODN were combined by the chemical spacer to increase its resistance to endonuclease for intratracheal administration. The therapeutic effect of chimeric decoy ODN was investigated using OVA-induced experimental asthma in mice. Mice were sensitized by intraperitoneal injections of 20 ug of ovalbumin (OVA) and 2 mg aluminum hydroxide constituted in 0.1 ml of saline on days 0 and 14. On days 21, 22 and 23, mice were challenged with 1% OVA aerosol for 20 min. At 24 hours after the last challenge, AHR was measured by methacholine-induced airflow obstruction. Intratracheal administration of decoy ODN was performed in OVA-sensitized mice at 3 days before the first challenge with aerosolized OVA. FITC-labeled chimeric decoy ODN could be detected in macrophages and monocytes migrating into the lung and airway, and NFkB and STAT6 activity were simultaneously inhibited by chimeric decoy ODN. Twenty-four hours after the last OVA challenge, treatment with chimeric or single transfection of NFkB decoy ODN was protected from methacholine-induced AHR, while mice treated with scrambled decoy ODN or saline developed a significantly increase in airway reactivity to methacholine. Importantly, this inhibitory effect of chimeric decoy ODN on airway hyperresponsiveness was significantly greater than that of NFkB decoy ODN. Treatment with chimeric decoy ODN markedly suppressed airway inflammation after OVA sensitization and challenge as compared with control and scrambled decoy ODN treatment. Inflammatory infiltrate, such as macrophage, was significantly inhibited by chimeric decoy ODN through suppression of ICAM-1 and eotaxin expression. In addition, secretion of Th2 cytokines including IL-4, IL-5 and IL-13 in BALF, and histamine in the whole lung were reduced by chimeric decoy ODN. Furthermore, a significant reduction of mucin secretion was observed by chimeric decoy ODN treatment accompanied by a suppression of MUC5AC gene expression. However, intratracheal administration of chimeric decoy ODN did not affected IgE synthesis. The present study provides a novel strategy for treating bronchial asthma by the simultaneous inhibition of both NFkB and STAT6 using chimeric decoy ODN. Further modification of chimeric decoy ODN would be useful to treat asthma as a decoy-based therapy.

Studies are underway for the development of cutoff sheets (CH-200) for use at final disposal yards, which are capable of preventing the expansion of perforations and of self-repair. The study reported here was concerned with the basic properties of these cutoff sheets, in terms of the effects of water pressure, temperature and quality, as well as the structure of the sheets themselves, on their capacity for self-repair. It was found that the capacity for self-repair could be maintained under water pressure of up to 3kgf/cm2, and at the temperatures normally encountered in groundwater and waste.

GATA3 mutations cause HDR (hypoparathyroidism, sensorineural deafness, and renal dysplasia) syndrome and, consistent with the presence of the second DiGeorge syndrome locus (DGS2) proximal to GATA3, distal 10p deletions often leads to HDR and DiGeorge syndromes. Here, we report on six Japanese patients with GATA3 abnormalities. Cases 1-5 had a normal karyotype, and case 6 had a 46,XX,del(10)(p15) karyotype. Cases 1-6 had two or three of the HDR triad features. Case 6 had no DiGeorge syndrome phenotype except for hypoparathyroidism common to HDR and DiGeorge syndromes. Mutation analysis showed heterozygous GATA3 mutations in cases 1-5, i.e., c.404-405insC (p.P135fsX303) in case 1, c.700T>C & c.708-709insC (p.F234L & p.S237fsX303) on the same allele in case 2, c.737-738insG (p.G246fsX303) in case 3, c.824G>T (p.W275L) in case 4, and IVS5+1G>C (splice error) in case 5. Deletion analysis of chromosome 10p revealed loss of GATA3 and preservation of D10S547 in case 6. The results are consistent with the previous finding that GATA3 mutations are usually identified in patients with two or three of the HDR triad features, and provide supportive data for the mapping of DGS2 in the region proximal to D10S547.

It is well-known that a mother wavelet for the discrete wavelet transform (DWT) has the band-pass filter characteristic with octave width in the frequency domain and can be used for octave analysis. However, it is possible that the octave analysis is not necessarily the most suitable to match the analysis signal. In this study, in order to construct the most suitable basis to match the analysis signal, a novel variable-filter band discrete wavelet transform (VFB-DWT) is proposed. It is achieved by using variable-band filters instead of conventional decomposition and reconstruction sequences, which are designed in consideration of the real signal characteristics. Additionally, it is proven that perfect reconstruction of the analysis signal by VFB-DWT is guaranteed using the perfect shift invariant theorem that underlies the theory of the PTI-CDWT having base DWT.

In this study, in order to verify the effectiveness of the variable filter band discrete wavelet transform (VFB-DWT) and construction method of the variable-band filter (VBF), a fetal ECG extraction has been carried out and the main results obtained are as follows. The approach to configuration VBF by selecting the frequency band only where the fetal ECG component is present was effective to configure the optimal base sensible signal. The extraction of the fetal ECG was successful by applying the wavelet shrinkage to VFB-DWT, which used the constructed VBF. The information entropy was selected as an evaluation index, and two kinds of ECG signals are used to evaluate the wavelet transform basis between the wavelet packet transform (WPT) and the VFB-DWT. One is a synthesized signal composed of white noise, the maternal ECG and the fetal ECG. The other signal is the real target signal separated by independent component analysis (ICA) and has the mother's body noise, the maternal ECG and the fetal ECG. The result shows that the basis by VBF of the VFB-DWT is better than the basis of the WPT that was chosen by the best basis algorithm (BBA).

In this paper, we propose a design method for directional selection in the two-dimensional complex wavelet packet transform (2D-CWPT). Current two-dimensional complex discrete wavelet transforms (2D-CDWT) can extract directional components from images, but the number of directions is small, and the directions and resolutions are fixed. Thus the current 2D-CDWTs are not flexible enough. In this study, we propose a new design method of the directional filters that can detect desirable direction components. Additionally flexible directional selection is achieved because the directional filters are added to the 2D-CWPT. Finally, the proposed method is applied to defect detection in semiconductor wafer circuits and an encouraging result is obtained.

With the aim of elucidating in greater detail the genealogical origin of the present domestic fowls of the world, we have determined mtDNA sequences of the D-loop regions for a total of 21 birds, of which 12 samples belong to red junglefowl (Gallus gallus) comprising three subspecies (six Gallus gallus gallus, three Gallus gallus spadiceus, and three Gallus gallus bankiva) and nine represent diverse domestic breeds (Gallus gallus domesticus). We also sequenced four green junglefowl (Gallus varius), two Lafayette's junglefowl (Gallus lafayettei), and one grey junglefowl (Gallus sonneratii). We then constructed a phylogenetic tree for these birds by the use of nucleotide sequences, choosing the Japanese quail (Coturnix coturnix japonica) as an outgroup. We found that a continental population of G. g. gallus was the real matriarchic origin of all the domestic poultries examined in this study. It is also of particular interest that there were no discernible differences among G. gallus subspecies; G. g. bankiva was a notable exception. This was because G. g. spadiceus and a continental population of G. g. gallus formed a single cluster in the phylogenetic tree. G. g. bankiva, on the other hand, was a distinct entity, thus deserving its subspecies status. It implies that a continental population of G. g. gallus sufficed as the monophyletic ancestor of all domestic breeds. We also discussed a possible significance of the initial dispersal pattern of the present domestic fowls, using the phylogenetic tree.

Further comparison of mitochondrial control-region DNA base sequences of 16 avian species belonging to the subfamily Phasianinae revealed the following: (i) Generalized perdicine birds (quails and partridges) are of ancient lineages. Even the closest pair, the common quail (Coturnix coturnix japonica) and the Chinese bamboo partridge (Bambusicola thoracica), maintained only 85.71% identity. (ii) The 12 species of phasianine birds previously and presently studied belonged to three distinct branches. The first branch was made exclusively of members of the genus Gallus, while the second branch was made of pheasants of the genera Phasianus, Chrysolophus, and Syrmaticus. Gallopheasants of the genus Lophura were distant cousins to these pheasants. The great argus (Argusianus argus) and peafowls of the genus Pavo constituted the third branch. The position of peacock-pheasants of the genus Polyplectron in the third branch was similar to that of the genus Lophura in the second branch. Members of the fourth phasianine branch, such as tragopans and monals, were not included in the present study. (iii) The one perdicine species, Bambusicola thoracica, was more closely related to phasianine genera Gallus and Pavo than to members of other perdicine genera. The above might indicate that Bambusicola belong to one-stem perdicine lineage that later splits into two sublineages that yielded phasianine birds, one evolving to Gallus, and the other differentiating toward Pavo and its allies.

The noncoding control region of the mitochondrial DNA of various gallinaceous birds was studied with regard to its restriction fragment length polymorphism (RFLP) and sequences of the first 400 bases. Tandem duplication of the 60-base unit was established as a trait unique to the genus Gallus, which is shared neither by pheasants nor by quails. Unlike its close ally Gallus varius (green junglefowl), the red junglefowl Gallus gallus is a genetically very diverse species; the 7.0% sequence divergence was seen between those from Thailand (G. g. gallus and G. g. spadiceus) and the other from the Indonesian island of Java (G. g. Bankiva). Furthermore, the divergence increased to 27.83% if each transversion is regarded as an equivalent of 10 transitions. On the other hand, a mere 0.5-3.0% difference (all transitions) separated various domestic breeds of the chicken from two G. g. gallus of Thailand, thus indicating a single domestication event in the area inhabited by this subspecies of the red junglefowl as the origin of all domestic breeds. Only transitions separated six diverse domesticated breeds. Nevertheless, a 2.75% difference was seen between RFLP type I breeds (White Leghorn and Nagoya) and a RFLP type VIII breed (Ayam Pelung). The above data suggested that although the mitochondrion of RFLP type V was the main contributor to domestication, hens of other RFLP types also contributed to this event.

The 1H-NMR spectra of native human epidermal growth factor (EGF) and a derivative lacking the final five residues have been assigned by two-dimensional methods, enabling their structures to be compared. The same structural features are observed for each protein, although the final five residues of native human EGF interact with residues earlier in the sequence. Comparison of the resonance shifts of human, rat and mouse EGF and human transforming growth factor alpha (TGF alpha) enables shifts characteristic of the EGF conformation to be identified, providing standards by which the structures of related proteins may be assessed.

Visualization of a three-dimensional fish behavior based on the successive feature extraction using the time sequential three-dimensional shape modeling is reported. The principle of the modeling is based on the intersected viewing pyramid consisting of each image boundary of the object and the corresponding focal point. Threedirectional video images are simultaneously acquired and sequential solid models are then reconstructed to obtain behavioral parameters such as body shape, skeleton, head, tail, gravity center, direction, attack angle, trajectory, velocity, acceleration, and moving energy. This procedure is entirely automatic requiring no manual operation and appears to have some considerable utility in tracking and measurement of moving objects.

Human insulin-like growth factor II (hIGF-II) was produced inEscherichia coli as a protein fused to human growth hormone. High level expression of the fusion protein was attained with pIBL-1 plasmid. The hIGF-II obtained byin vitro cleavage of the fusion protein with cyanogen bromide was highly purified and its biological activity was assessed.

A RNA molecule has been synthesized that is identical in sequence to Escherichia coli tRNAfMet except that it lacks the base modifications present in the E. coli tRNA. This was achieved by enzymatic joining of chemically synthesized oligonucleotides with chain lengths of 3-10 which were synthesized by the phosphodiester or phosphotriester method. First, quarter molecules of tRNA were constructed by joining of chemically synthesized fragments with RNA ligase. The 5'-quarter molecule (bases 1-20) served as an acceptor in joining reactions with the 3',5'-bisphosphorylated donor molecule (bases 21-34). The 5'-half molecule thus obtained was treated with phosphatase and joined to the 3'-half molecule which was prepared by ligation of the other quarter molecules (bases 35-60, acceptor; bases 61-77, donor) followed by 5'-phosphorylation with polynucleotide kinase. The synthetic tRNA was characterized by oligonucleotide pattern and was partially active in aminoacylation with E. coli methionyl-tRNA synthetase.

Endeavoring to develop a method to biosynthesize proteins substituted with nonprotein amino acids, we attempted the incorporation of L-2-aminohexanoic acid (Ahx) into human epidermal growth factor (hEGF). Escherichia coli YK537 strain harboring plasmid pTA1522, which has the phoA promoter-phoA signal peptide-hEGF gene, was used. Cells were cultured first in high-phosphate medium and then, for induction of the hEGF-encoding gene, transferred to low-phosphate medium containing Ahx (0.25 mg/ml). hEGF and Ahx-substituted hEGF, [Ahx21]hEGF, secreted into the periplasm were recovered. After treatment with H2O2, [Ahx21]-hEGF was clearly separated from methionine-oxidized hEGF by one-step reverse-phase HPLC. Substitution of the methionine residue of hEGF with Ahx was confirmed by the amino acid analysis of [Ahx21]hEGF. The three biological activities of [Ahx21]hEGF were the same as those of hEGF. From the successful production of [Ahx21]hEGF, a basic strategy was established for preparing proteins substituted with nonprotein amino acid (alloprotein). Induction of the phoA promoter of pho regulon and secretion of the product to the periplasm may depress heat shock-like responses and subsequent hydrolysis of the product by cytoplasmic protease.

Oligonucleotides corresponding to the E. coli tRNA (f)(Met) have been synthesized. A diester approach has been used for the synthesis of 6 , 7 , 8 and 9 . A block condensation of nucleotides with 3′-phosphodiesters has been applied to the synthesis of 2 , 3 , 4 and 5 . A new phosphorylating reagent, p-chlorophenyl N-phenylphosphoroamido chloride was synthesized and used for this purpose. Joining of previously synthesized fragments with T4 RNA ligase yielded an eicosanucleotide ( 1 + 2 ) corresponding to bases 1–20 from the 5′-end.

A tuft in the air flow was observed by three CCD video cameras which were installed in the directions nearly perpendicular to each other. In this procedure, a tufted woolen yarn of the diameter of about 1 mm and of the length of 35 mm was attached on the top of a thin post pin.Principle of this shape modelling is that the intersected zone which is constructed from multiple polygonal pyramids, every of which is bounded by the digitized cross sectional area in the picture image of the target and its lens center as a vertex results a polyhedron and becomes a good approximation to that shape if it is convex. This polyhedron is designed to construct a so-called solid model and is described with spatially fixed co-ordinates, so that not only its spatical shape, but also the position, direction, deformation or fluttering at the every moment are able to be estimated.From the reconstructed shape modelling, influence of rigidness due to adhesives near the fixed end and a few millimeters of three dimensional displacement at the free end are observed. Free end moves like a cone of which vertex locates at the middle of the tuft.This method is capable to serve for measurement and analysis of a single tuft characteristics depending on its material, kind of fluid or flow range. And also this has the posibility of quantitative analysis of three dimensional unsteady flow visualization.

A hexanucleotide and an octanucleotide corresponding to tRNAMetf bases 41-46 and 47-54 were synthesized by stepwise addition of mononucleotides. The octanucleotide is the largest oligoribonucleotide synthesized by this method. A pentanucleotide (bases 61-65) and a hexanucleotide (bases 66-71) were synthesized via partially triesterified intermediates. The deblocked products were purified by ion-exchange chromatography and characterized by enzymatic hydrolysis. These oligonucleotides were used as substrates in joining reactions with RNA ligase.

Various protecting groups were introduced to the phosphate of 2'-O-benzoyluridine 3'-phosphate. Stability and selective removal of these groups from the trisubstituted phosphate were investigated and nucleotides with trisubstituted phosphate were uesd for the synthesis of dinucleotides by condensation with a properly protected mononucleotide.

A synthetic gene for porcine secretin was ligated with Pst I-cleaved pBR322 by its flanking synthetic Pst I linkers to produce a fused protein consisting of an amino-terminal portion of beta-lactamase and an entire secretin molecule. The hybrid plasmid was transferred into competent Escherichia coli cells. The plasmids, which were proved in our investigation to contain the secretin gene in the desired orientation, were then screened for the production of secretin. This was shown by radioimmunoassay and gel electrophoretic analysis of the polypeptides that were synthesized in E. coli minicells transformed with the hybrid plasmids. Secretin so produced, whose carboxy terminal residue may not be amidated in contrast with natural porcine secretin, showed the same activity in stimulating pancreatic secretion in a bioassay with anesthetized rats as does natural secretin.

P1-Adenosine 5'-P2-o-nitrobenzyl pyrophosphate (nbzlppA) has been synthesized as a substrate for T4 RNA ligase catalyzed 3'-phosphorylation. Incubation of oligoribonucleotides and nbzlppA with RNA ligase yielded oligoribonucleotides having a 3'-o-(o-nitrobenzyl) phosphate. Photochemical removal of the o-nitrobenzyl group provided the free 3'-phosphate. Using [P2-32P] nbzlppA, 3'-termini of oligoribonucleotides could be labelled with 32P. This reaction was applied to modify the 3'-end of donor molecules in joining reaction with RNA ligase. A trinucleotide U-A-G was converted to U-A-Gpnbzl and phosphorylated with polynucleotide kinase. pU-A-Gpnbzl was then joined to an acceptor trinucleotide A-U-G to yield A-U-G-U-A-Gp.