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41.
Identification,cDNA cloning and possible roles of seed-specific rice asparaginyl endopeptidase,REP-2
We previously showed that two major cysteine endopeptidases, REP-1 and REP-2, were present in germinated rice ( Oryza sativa L.) seeds, and that REP-1 was the enzyme that digests seed storage proteins. The present study shows that REP-2 is an asparaginyl endopeptidase that acts as an activator of REP-1, and we separated it into two forms, REP-2alpha (39 kDa) and REP-2beta (40 kDa), using ion-exchange chromatography and gel filtration chromatography. Although analysis of the amino terminals revealed that 10 amino acids of both forms were identical, their isoelectric points were different. SDS-PAGE/immunoblot analysis using an antiserum raised against legumain, an asparaginyl endopeptidase from jack bean, indicated that both forms were present in maturing and germinating rice seeds, and that their amounts transiently decreased in dry seeds. Northern blot analysis indicated that REP-2 mRNA was expressed in both maturing and germinating seeds. In germinating seeds, the mRNA was detected in aleurone layers but not in shoot and root tissues. Incubation of the de-embryonated seeds in 10(-6) M gibberellic acid induced the production of large amounts of REP-1, whereas REP-2beta levels declined rapidly. Southern blot analysis showed that there is one gene for REP-2 in the genome, indicating that both REP-2 enzymes are generated from a single gene. The structure of the gene was similar to that of beta-VPE and gamma-VPE isolated from Arabidopsis thaliana. 相似文献
42.
Darsaud A Bourdon L Chevrier C Keita M Bouteille B Queyroy A Canini F Cespuglio R Dumas M Buguet A 《Experimental biology and medicine (Maywood, N.J.)》2003,228(11):1355-1362
Animal models of Human African Trypanosomiasis (HAT) have been developed to understand the pathogenic mechanisms leading to the passage into the neurological phase, most of them referring to histological aspects but not clinical or behavioral data. Our study aimed at defining simple clinical and/or behavioral markers of the passage between the hemolymphatic phase and the meningo-encephalitic stage of the disease. Sprague-Dawley rats (n=24) were infected with Trypanosoma brucei brucei AnTat 1.1E. Food intake and body weight were measured daily from the day of infection until death. Hematocrit was measured twice a week. Behavioral disturbances were evaluated through an Open-field test. A sudden weight loss occurred on the twelfth day after infection, due to a significant drop of food intake starting two days before. The rats developed an anemic state shown by the hematocrit measurements. The Open-field test showed them to be less active and reactive as soon as the second week after infestation. A complementary histological study observed trypanosomes and inflammatory cells in the choroid plexus at the same period. These results are in favor of central nervous system functional disturbances. The observed weight loss is discussed as being a parameter of the entry in the meningo-encephalitic phase. The rat model reproduces neurological symptoms observed in the human disease and may prove to be useful for further neurohistological and therapeutic studies. 相似文献
43.
Naoya Yoshihara Taiji Sakamoto Takehiro Yamashita Toshifumi Yamashita Keita Yamakiri Shozo Sonoda Tatsuro Ishibashi 《PloS one》2015,10(4)
Purpose
To determine whether the width of the retinal artery (RA) trajectory was associated with the presence of a macular hole (MH).Methods
A retrospective cross sectional case-control study was performed. The fundus photographs were rotated 90 degrees, and the coordinates of the best fit curve of the RA trajectory were determined automatically based on these plots using the ImageJ program. The converted coordinates were fit to a second degree polynomial (ax2/100 + bx + c) equation. The width and steepness of the RA trajectory, “a”, of the eyes with a MH eye were compared to that of the fellow eyes.Results
One hundred and ten eyes of 55 consecutive patients (30 women) with a unilateral MH and healthy fellow eyes were analyzed. The mean age was 64.9 years (range 47-81 years). The constant ‘a’ was significantly smaller in eyes with a MH than that of the fellow eyes (0.379 ± 0.094 vs 0.416 ± 0.121, P = 0.001, paired t test), indicating that the RA trajectory was wider in the MH eyes than in the fellow eyes. There was a significant correlation between the axial length and ‘a’ of the RA trajectory in the MH eyes (R = 0.273, P = 0.044) and in the fellow eyes (R = 0.356, P = 0.008; Spearman’s rank correlation coefficient).Conclusions
Because eyes with a MH have a significantly wider and flatter RA trajectory, there may be greater traction on the fovea which is located between the RA arches. The causative role of this finding is still unclear. 相似文献44.
45.
Nobukatsu Horita Kiichiro Tsuchiya Ryohei Hayashi Keita Fukushima Shuji Hibiya Masayoshi Fukuda Yoshihito Kano Tomohiro Mizutani Yasuhiro Nemoto Shiro Yui Ryuichi Okamoto Tetsuya Nakamura Mamoru Watanabe 《Biochemical and biophysical research communications》2014
Background and aims
The dynamics of intestinal stem cells are crucial for regulation of intestinal function and maintenance. Although crypt stem cells have been identified in the intestine by genetic marking methods, identification of plural crypt stem cells has not yet been achieved as they are visualised in the same colour.Methods
Intestinal organoids were transferred into Matrigel® mixed with lentivirus encoding mCherry. The dynamics of mCherry-positive cells was analysed using time-lapse imaging, and the localisation of mCherry-positive cells was analysed using 3D immunofluorescence.Results
We established an original method for the introduction of a transgene into an organoid generated from mouse small intestine that resulted in continuous fluorescence of the mCherry protein in a portion of organoid cells. Three-dimensional analysis using confocal microscopy showed a single mCherry-positive cell in an organoid crypt that had been cultured for >1 year, which suggested the presence of long-lived mCherry-positive and -negative stem cells in the same crypt. Moreover, a single mCherry-positive stem cell in a crypt gave rise to both crypt base columnar cells and transit amplifying cells. Each mCherry-positive and -negative cell contributed to the generation of organoids.Conclusions
The use of our original lentiviral transgene system to mark individual organoid crypt stem cells showed that long-lived plural crypt stem cells might independently serve as intestinal epithelial cells, resulting in the formation of a completely functional villus. 相似文献46.
The function of GABA in the adrenal medulla is still controversial. We will review experimental results in vivo and in vitro
in adrenal chromaffin cells of various mammals to clarify what has been elucidated and what still remains to be settled. 相似文献
47.
Toshihiro Umebayashi Yasuhiro Utsumi Shinya Koga Susumu Inoue Junji Matsumura Kazuyuki Oda Seizo Fujikawa Keita Arakawa Kyoichi Otsuki 《Trees - Structure and Function》2010,24(3):571-583
A dye injection method was used to elucidate the xylem water-conducting pathways of 34 broadleaved evergreen trees growing
in southern Japan: two semi-ring-porous, 26 diffuse-porous, five radial-porous and one non-vessel species. The large earlywood
vessels in semi-ring-porous species have a water transport function in only the outermost annual ring, as in deciduous ring-porous
species. On the other hand, the small vessels in semi-ring-porous species maintain the water transport function in many outer
annual rings. For the other xylem-type species, the many vessels in many outer annual rings have a water transport function.
In diffuse-porous species, we categorized the water-conducting pattern within the annual rings into two types: d1 type, where
water travels through vessels in the whole region; and d2 type, where water travels mainly through the earlywood vessels.
The pattern in radial-porous species is similar to that in the d1 type; the pattern in non-vessels species is similar to that
in the d2 type. The vessel diameter in radial-porous species is similar to that of the earlywood vessels of semi-ring-porous
species. These results suggest that the conduit diameter size is only one of many factors determining the water-conducting
pathways of broadleaved evergreen species. 相似文献
48.
49.
50.
Ichitani K Namigoshi K Sato M Taura S Aoki M Matsumoto Y Saitou T Marubashi W Kuboyama T 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2007,114(8):1407-1415
Hybrid weakness is a reproductive barrier that is found in many plant species. In rice, the hybrid weakness caused by two complementary genes, Hwc1 and Hwc2, has been surveyed intensively. However, their gene products and the molecular mechanism that causes hybrid weakness have remained unknown. We performed linkage analyses of Hwc1, narrowed down the area of interest to 60 kb, and identified eight candidate genes. In the F(2) population, in which both Hwc1 and Hwc2 genes were segregated, plants were separable into four classes according to their respective phenotypes: severe type, semi-severe type, F(1) type, and normal type. Severe type plants show such severe symptoms that they could produce only tiny shoot-like structures; they were unable to generate roots. Genetic analyses using closely linked DNA markers of the two genes showed that the symptoms of the F(2) plants were explainable by the genotypes of Hwc1 and Hwc2. Weakness was observed in plants that have both Hwc1 and Hwc2. In Hwc1 homozygote, the symptoms worsened and severe type or semi-severe type plants appeared. Consequently, Hwc1 should have a gene dosage effect and be a semi-dominant gene. The dosage effect of Hwc2 was recognizable, but it was not so severe as that in Hwc1. These results are useful to elucidate the mechanism that causes the hybrid weakness phenomenon and the role of each causal gene in hybrid weakness. 相似文献