Biomedical subjects
Publications and source records attributed to H Noji.
We describe a 92-year-old male patient with systemic lupus erythematosus (SLE) who had sudden onset of thrombocytopenia and developed acute lupus pneumonitis (ALP). Although steroid pulse therapy was effective for ALP, he developed complicated bacterial pulmonary disease. This patient is the oldest ever reported to have contracted SLE.
The long period of external fixation after the completion of distraction, which is necessary to obtain enough strength for resisting without fixation is a significant disadvantage of distraction osteogenesis. The purpose of this study is to understand the mechanical property of the regenerated bone and try to find an appropriate timing for safely removing the fixation device. An external fixation device was applied to the right tibia in rabbits and transverse osteotomy was performed just below the tibio-fibula junction. The tibiae were lengthened 7.2 mm at 0.72 mm a day for 10 days after surgery. A bone mineral density (BMD) and stress strain index (SSI) analyzed by peripheral quantitative computed tomography showed a significant increase on day 40, but returned to the control level at day 64 after the completion of distraction. Therefore, the newly formed bone between the host cortical bone seemed to be enough to resist the mechanical stress on day 40 during the consolidation period in this study. The present results suggested the possibility of removing the fixation device during the period when the BMD and SSI showed a significantly high level during the consolidation period.
Homogeneous preparations of alpha(3)beta(3)gamma complexes with one, two or three non-competent non-catalytic site(s) were performed as described [Amano, Hisabori, Muneyuki, and Yoshida (1996) J. Biol. Chem. 271, 18128-18133] and their properties were compared with those of the wild-type complex. The ATPase activity of the complex with three non-competent non-catalytic sites decayed rapidly to an inactivated state, as reported previously [Matsui, Muneyuki, Honda, Allison, Dou, and Yoshida (1997) J. Biol. Chem. 272, 8215-8221]. In contrast, the complex with one or two non-competent non-catalytic sites displayed a substantial steady-state phase activity depending on the number of non-competent non-catalytic sites in the complex. This result indicates that one competent non-catalytic site can maintain the continuous catalytic turnover of the enzyme and can potentially relieve all three catalytic sites from inhibition by MgADP(-). Furthermore, the results suggest that the interaction between three non-catalytic sites might not be as strong as that between catalytic sites, which are all strictly required for a continuous catalytic turnover.
By applying the same method used for F(1)-ATPase (TF(1)) from thermophilic Bacillus PS3 (Noji, H., Yasuda, R., Yoshida, M., and Kinosita, K., Jr. (1997) Nature 386, 299-302), we observed ATP-driven rotation of a fluorescent actin filament attached to the gamma subunit in Escherichia coli F(1)-ATPase. The torque value and the direction of the rotation were the same as those observed for TF(1). F(1)-ATPases seem to share common properties of rotation irrespective of the sources.
In the crystal structure of mitochondrial F1-ATPase, two beta subunits with a bound Mg-nucleotide are in "closed" conformations, whereas the third beta subunit without bound nucleotide is in an "open" conformation. In this "CCO" (beta-closed beta-closed beta-open) conformational state, Ile-390s of the two closed beta subunits, even though they are separated by an intervening alpha subunit, have a direct contact. We replaced the equivalent Ile of the alpha3beta3gamma subcomplex of thermophilic F1-ATPase with Cys and observed the formation of the beta-beta cross-link through a disulfide bond. The analysis of conditions required for the cross-link formation indicates that: (i) F1-ATPase takes the CCO conformation when two catalytic sites are filled with Mg-nucleotide, (ii) intermediate(s) with the CCO conformation are generated during catalytic cycle, (iii) the Mg-ADP inhibited form is in the CCO conformation, and (iv) F1-ATPase dwells in conformational state(s) other than CCO when only one (or none) of catalytic sites is filled by Mg-nucleotide or when catalytic sites are filled by Mg2+-free nucleotide. The alpha3beta3gamma subcomplex containing the beta-beta cross-link retained the activity of uni-site catalysis but lost that of multiple catalytic turnover, suggesting that open-closed transition of beta subunits is required for the rotation of gamma subunit but not for hydrolysis of a single ATP.
We investigated the complement sensitivity of erythrocytes from three patients, one with inherited complete deficiency of CD59, one with the Inab phenotype, and one with paroxysmal nocturnal haemoglobinuria (PNH). The complement lysis sensitivity units on the erythrocytes were 11.7, 4.6, and 47.6 for inherited CD59 deficiency, Inab phenotype, and PNH, respectively. Two-colour flow cytometric analysis showed that the erythrocytes from the three patients consisted of a single population negative for CD59, negative for decay accelerating factor (DAF), and negative for both proteins, respectively. In addition, only the Inab phenotype patient had no haemolysis in vivo. These facts suggest that CD59 deficiency plays a more important role than DAF deficiency in complement-mediated haemolysis in vitro and in vivo, and that deficiency of both proteins, but not CD59 or DAF alone, causes complement sensitivity corresponding to that of PNH III erythrocytes in vitro.
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A 25-year-old man was admitted for evaluation of pancytopenia on May 2, 1997. On admission, he had pancytopenia with a normal reticulocyte count. Bone marrow aspirate specimens displayed a normal karyotype and hypocellularity without myelodysplasia. Although total bilirubin and lactate dehydrogenase levels were within their normal ranges, the haptoglobin level was low; additionally, two-color flow cytometric analysis determined that 3.3% of erythrocytes were double-negative for CD55 and CD59 expression. Atypical paroxysmal nocturnal hemoglobinuria with bone marrow hypoplasia was diagnosed. Because initial treatment with cyclosporin A was not effective, the patient was subsequently given a combination of antithymocyte globulin, cyclosporin A, and granulocyte colony-stimulating factor. Although the pancytopenia subsided, the percentage of double-negative erythrocytes in the patient's blood remained almost unchanged compared to findings obtained on admission.
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Rotation of the epsilon subunit in F1-ATPase from thermophilic Bacillus strain PS3 (TF1) was observed under a fluorescence microscope by the method used for observation of the gamma subunit rotation (Noji, H., Yasuda, R., Yoshida, M., and Kinosita, K., Jr. (1997) Nature 386, 299-302). The alpha3 beta3 gamma epsilon complex of TF1 was fixed to a solid surface, and fluorescently labeled actin filament was attached to the epsilon subunit through biotin-streptavidin. In the presence of ATP, the filament attached to epsilon subunit rotated in a unidirection. The direction of the rotation was the same as that observed for the gamma subunit. The rotational velocity was slightly slower than the filament attached to the gamma subunit, probably due to the experimental setup used. Thus, as suggested from biochemical studies (Aggeler, R., Ogilvie, I. , and Capaldi, R. A. (1997) J. Biol. Chem. 272, 19621-19624), the epsilon subunit rotates with the gamma subunit in F1-ATPase during catalysis.
A single molecule of F1-ATPase, a portion of ATP synthase, is by itself a rotary motor in which a central gamma subunit rotates against a surrounding cylinder made of alpha3beta3 subunits. Driven by three catalytic betas, each fueled with ATP, gamma makes discrete 120 degree steps, occasionally stepping backward. The work done in each step is constant over a broad range of imposed load and is close to the free energy of hydrolysis of one ATP molecule.
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Rapid demographic aging has made caring for the elderly an increasingly important social issue in Japan. To study current conditions of the oldest elderly citizens, we investigated the dietary practices of centenarians in the Tokyo metropolitan area. First, we compared the food intake of centenarians with that of octogenarians. Next, to identify dietary trends, we investigated whether food intake by centenarians had changed significantly between 1981 and 1995. Nutritional intake by the centenarians and octogenarians in 1995 was about 60% and 75% that of the control, respectively. However, the nutritional intake of well nourished centenarians was similar to that of the octogenarians. Cognitive function and daily activity have an influence on nutritional intake. The centenarians were similar to the control subjects in their consumption of dairy products, sweets, and fruit. However, their intake of cereals, meat, fish, and fatty oils was loss than 60% that of the control, which indicates their preference for soft and sugary foods. The pattern of dietary practices of centenarians in 1981 was similar. Although the total food intake of centenarians amounted to 60% of the control in 1995 energy intake per kilogram of body weight averaged over 30 kcal. As to dietary trends, centenarians in 1981 are more cereals, eggs, algae products, and legumes than did their 1995 counterparts. This finding seems to reflect a generational difference in dietary habits.
To clarify the response of endothelial cells to complement, we studied not only the reaction of endothelial cells against complement lysis sensitivity (CLS) test, but also the expression of complement regulatory proteins by two-color flow cytometric analysis and backscattered scanning immunoelectron microscopic analysis using monoclonal antibodies to decay-accelerating factor (DAF) and/or CD59. Complement activation didn't lead to the cell death of human aortic endothelial cells (HAECs) and human umbilical vein endothelial cells (HUVECs). In control, two-color flow cytometric analysis indicated that HAECs consisted of a single double-positive population for these proteins as well as HUVECs. Then, HUVECs and HAECs after the CLS test resulted in having same distribution by flow cytometry. Moreover, the microscopic analysis showed that DAF or CD59 was expressed with a diffuse distribution. However, DAF on HUVECs and CD59 on HAECs were present at the margin of cell surfaces more than at the other places. These findings suggest that endothelial cells have a defense mechanism for complement activation in vitro by the changes of expression of complement regulatory proteins on the membrane surface, and that the mechanism of HAECs to complement is the same as that of HUVECs.
Cells employ a variety of linear motors, such as myosin, kinesin and RNA polymerase, which move along and exert force on a filamentous structure. But only one rotary motor has been investigated in detail, the bacterial flagellum (a complex of about 100 protein molecules). We now show that a single molecule of F1-ATPase acts as a rotary motor, the smallest known, by direct observation of its motion. A central rotor of radius approximately 1 nm, formed by its gamma-subunit, turns in a stator barrel of radius approximately 5nm formed by three alpha- and three beta-subunits. F1-ATPase, together with the membrane-embedded proton-conducting unit F0, forms the H+-ATP synthase that reversibly couples transmembrane proton flow to ATP synthesis/hydrolysis in respiring and photosynthetic cells. It has been suggested that the gamma-subunit of F1-ATPase rotates within the alphabeta-hexamer, a conjecture supported by structural, biochemical and spectroscopic studies. We attached a fluorescent actin filament to the gamma-subunit as a marker, which enabled us to observe this motion directly. In the presence of ATP, the filament rotated for more than 100 revolutions in an anticlockwise direction when viewed from the 'membrane' side. The rotary torque produced reached more than 40 pN nm(-1) under high load.
ATP-dependent, azide-sensitive rotation of the gamma subunit relative to the alpha3beta3 hexagonal ring of ATP synthase was observed with a single molecule imaging system. Thus, ATP synthase is a rotary motor enzyme, the first ever found.
To establish the guideline for nutrition in the very old, we analyzed biochemical and hematological data from 45 Japanese centenarians living in Tokyo metropolitan area during 1994 and 1995. Levels of cholesterol, HDL-cholesterol, apolipoprotein A1 and B, albumin, prealbumin in serum were lower than in a control group or lower than the reference range, which indicates that these centenarians were undernourished. RBC counts, hemoglobin, and hematocrit were also low in the centenarians. The concentration of albumin correlated positively with those of HDL-cholesterol and apolipoprotein A1, and negatively with concentration of Lp(a). The value for transferrin correlated positively with energy intake and with the concentration of apolipoprotein A1. The concentration of prealbumin correlated positively with the concentration of hemoglobin, hematocrit and the total cholesterol concentration. According to their nutritional status, the centenarians were divided into 2 groups: well-nourished and undernourished. Those who were well-nourished had higher levels of cholesterol, HDL, Hb, and apolipoprotein A1, their levels of ADL and cognitive function were also high. The findings of blood-chemical data in the centenarians may be partly due to undernutrition. Centenarians who were well-nourished were considered to be aging successfully. These results and others previously published indicate that the concentrations of albumin, prealbumin, transferrin and either CRI or IL-6 are useful for nutritional assessment in the very old.
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