cDNA cloning, tissue distribution, and subcellular localization of horseshoe crab big defensin

A full-length cDNA for horseshoe crab big defensin with a strong antimicrobial activity was obtained from a hemocyte cDNA library. The open reading frame of the cDNA coded for an NH2-terminal signal sequence followed by a propeptide and the mature big defensin. The propeptide is linked to the mature protein through an -Arg-X-Lys/Arg-Arg- motif, the processing site for Kex2-like proteases. Northern blot analysis revealed that big defensin is expressed in all the tissues tested, suggesting that big defensin plays an important role not only in hemocytes but also in other tissues for host defense. The subcellular localization, determined by immunocytochemistry at ultrastructural level, confirmed the previous findings obtained by biochemical analysis that big defensin locates in both small and large granules in hemocytes. Big defensin is the first example to demonstrate the existence of broad tissue distribution in horseshoe crab.     

Development of spectroscopic transmission-type four detector polarimeter

We have developed a spectroscopic transmission-type four detector polarimeter (T-FDP). It consists of a detector head and a multichannel spectrometer equipped with a two-dimensional CCD detector. Inside the T-FDP, three cubic beam splitters are aligned in a straight line and they are rotated relative to each other. From the responses of the spectroscopic T-FDP to five inputs with known polarization states it is possible to determine the characteristic matrices of the T-FDP at various wavelengths. The trajectories of the experimentally measured polarization states on the Poincaré sphere agree well with theoretical predictions. These results demonstrate the feasibility of using the T-FDP for spectroscopic ellipsometry.     

Elevated serum lipoprotein (a) levels associated with ulcerative colitis in a young Japanese patient

Thromboembolism has been shown to play a role in the pathogenesis of inflammatory bowel disease (IBD). A possibility exists that lipoprotein (a) [Lp(a)], a newly-discovered prothrombotic factor, also participates in the development of at least some cases of IBD. Marked elevation of serum Lp(a) levels was observed in a young patient with ulcerative colitis. A biopsy specimen of the rectal mucosa showed findings compatible with ulcerative colitis, as well as small vessel thrombus occurring within the muscularis mucosa in the rectum. Serum Lp(a) levels were markedly elevated on admission (71 mg/dl), with a gradual decrease to 46 mg/dl on discharge. Moreover, serum Lp(a) levels decreased in parallel with clinical improvement. In the quiescent clinical stage, no small vessel thrombus was observed in the mucosa on follow-up colonoscopy. The association between IBD and hyper-Lp(a)-emia would be presumable but it has been, to our knowledge, previously unreported. The case reported here would be the first young patient, suggesting the presence of hyper-Lp(a)-emia and small vessel thrombus formation occurring in association with the development of ulcerative colitis.     

A xanthine oxidase inhibitor 1'-acetoxychavicol acetate inhibits azoxymethane-induced colonic aberrant crypt foci in rats

The modifying effect of dietary administration of a xanthine oxidase inhibitor 1'-acetoxychavicol acetate (ACA) present in an edible plant Languas galanga in Thailand on the development of azoxymethane (AOM)-induced colonic aberrant crypt foci (ACF) was investigated in rats. Male F344 rats were given s.c. injections of AOM (15 mg/kg body wt) once a week for 3 weeks to induce colonic ACF. They were fed the diets containing 100 or 200 ppm ACA for 5 weeks, starting 1 week before the first dosing of AOM. At the termination of the study (week 5), AOM induced 118 +/- 28 ACF/colon. Dietary administration of ACA caused significant reduction in the frequency of ACF (41% inhibition by 100 ppm ACA feeding and 37% inhibition by 200 ppm ACA feeding, P<0.01). Such inhibition might be associated with suppression of the proliferation biomarkers' expression such as ornithine decarboxylase activity in the colonic mucosa, number of silver-stained nucleolar organizer regions' protein in the colonic mucosal cell nuclei and blood polyamine content. These results indicate that ACA could inhibit the development of AOM-induced ACF through its suppression of cell proliferation in the colonic mucosa and ACA might be a possible chemopreventive agent against colon tumourigenesis.     

Intratumoral pharmacokinetics and in vivo gene expression of naked plasmid DNA and its cationic liposome complexes after direct gene transfer

The pharmacokinetic properties and gene expression of naked plasmid DNA and its cationic liposome complexes were studied after direct intratumoral injection. Using a Walker 256 tissue-isolated tumor perfusion system, we quantified the recovery of naked plasmid DNA and cationic liposome complexes in the tumor, leakage from the tumor surface, and the venous outflow after intratumoral injection. Approximately 50% of naked plasmid DNA had been eliminated from the tumor 2 h after injection, whereas more than 90% of plasmid DNA was retained in the tumor when it was complexed with cationic liposomes. However, the distribution of these complexes in the tumor was restricted to the tissue surrounding the injection site. Pharmacokinetic analysis of the venous outflow profiles suggested that the rate-limiting process that determines the retention of plasmid DNA in the tumor is transferred from the injection site in the tumor tissue and that complexation with cationic liposomes may retard this process. Furthermore, we examined the gene expression of chloramphenicol acetyltransferase DNA constructs (naked pCMV-CAT) and the corresponding cationic liposome [3-beta-(N-(N',N'-dimethylaminoethane)carbamoyl)cholesterol] complexes. A similar level of gene expression was observed in vivo after direct intratumoral injection of naked DNA and its cationic liposome complexes. In both cases, a great variation was observed between tumors, and localization of gene-transduced cells in the tumor tissue was limited to the area in the vicinity of the injection site. Thus, these pharmacokinetic and gene expression studies have demonstrated that cationic liposomes can enhance the retention of injected DNA in the tumor model, whereas cationic liposome complex does not necessarily improve gene expression because of its poor dissemination in this tumor. The present study also suggested that there is a need to control the behavior of the injected naked plasmid DNA and its cationic liposome complexes to ensure better distribution throughout the tumor.     

Switching of H∞ controller under constraints and decision of controller state at instance of the controller switching

In real control systems, certain constraints are placed on the input, state, and output values. If such constraints are violated, the system may become unstable, in a worst‐case scenario. One of the promising approaches to solving this problem is to switch the controller according to the state of the closed‐loop system. No useful design strategy for each controller, however, has yet been developed. In addition, the determination of the controller state at the instant of controller switching has not been investigated. First, this paper presents a new design methodology which merges the switching control and H_∞ controller design into one design algorithm. Second, this paper presents a way of determining the controller state at the instant of controller switching. Finally, the proposed method is applied to force control of a one‐DOF manipulator in order to demonstrate the usefulness of the proposed method. © 2002 Wiley Periodicals, Inc. Electr Eng Jpn, 142(2): 68–75, 2003; Published online in Wiley InterScience ( www.interscience.wiley.com ). DOI 10.1002/eej.10082     

Direct fractionation of proteins in particle-containing feedstocks by a filter paper pieces-based DEAE-cellulose column chromatography. Rapid, robust and low-cost capturing procedure for protein

Goldfish were classically conditioned to a mixture of two pulse trains differing in both repetition rate and the spectral profile of the pulses. Animals were then tested for generalization to single pulse trains having one or the other spectral profile presented at a variety of repetition rates. Generalization functions of repetition rate were qualitatively similar to those obtained following conditioning to either of the pulse trains alone. Thus, the spectral profile of each pulse type was appropriately associated with the repetition rate at which that pulse type was presented during conditioning. These results indicate that the two concurrent pulse trains making up the conditioning stimuli were analyzed independently, forming two auditory streams. When either of the two pulse trains were presented with a 500 ms onset asynchrony, stream segregation was enhanced. These and other results suggest that many fundamental features of the human sense of hearing are widely shared among vertebrate animals, and may have developed first among fishes.     

Sex differences in oxidative damage in ddY mouse kidney treated with a renal carcinogen, iron nitrilotriacetate

Iron-induced free radical injuries in male and female ddY mice, especially the sex difference and its mechanisms, were studied after an i.p. injection of a renal carcinogen, ferric nitrilotriacetate. Male mice were much more susceptible to iron-induced free radical injuries than female mice. Oxidative modification of proteins and DNA occurred more strongly in males than in females, as measured by protein carbonyl content and 8-hydroxydeoxyguanosine, respectively. Histochemical detection of 4-hydroxy-2-nonenal-modified proteins using an antibody and DNA fragmentation as detected by the TUNEL method also showed that males are more severely damaged than females, especially in the proximal convoluted tubules. These results could not be explained by the difference in iron status between male and female mice. In fact, the toxic so-called 'free' iron in serum and kidney were not different between male and female mice and storage iron, such as ferritin and hemosiderin, was also comparable in both kidneys. In previous studies we proposed the glutathione cycling hypothesis to explain the sex differences. The half-life of glutathione in the kidney was significantly shorter in males (29 min) than in females (57 min), as determined by the glutathione decrease after buthionine sulfoximine treatment, a specific inhibitor of glutathione synthesis. The specific activity of gamma-glutamyltranspeptidase (EC 2.3.2.2) in female mice was 73% of that in male mice. These results suggest that the faster glutathione turnover in males could account for the higher susceptibility to oxidative injury by supplying the reducing equivalent that reduces Fe(III) to Fe(II), thereby facilitating iron-catalyzed free radical reactions.     

Differences in muscle and fat accretion in Japanese Black and European cattle

The development of different muscles and adipose tissues during growth was investigated in commercial Japanese Black (JB) cattle and compared with breeds of the largest variation to be found in Europe. Animals, reared under typical conditions for Japanese and European beef production systems, gained similar body weights but different carcass composition at 24 months of age. The carcass of JB contained more adipose tissue and the least proportion of muscle. The longissimus muscle of JB developed extraordinary amounts of 23.3% intramuscular fat (IMF) at 24 months of age, compared from 0.6% to 4.7% in European breeds. The relationships between IMF content in the longissimus muscle and different adipose tissue weights indicate that a large amount of “waste fat” is accreted with every percent of IMF. However in JB, the good ability of IMF deposition is associated with relatively least development of “waste fat”, as a result of unique breed characteristics combined with special feeding system.