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Yang, T.

Publications and source records attributed to Yang, T..

Hydrogen peroxide homeostasis: activation of plant catalase by calcium/calmodulin

Environmental stimuli such as UV, pathogen attack, and gravity can induce rapid changes in hydrogen peroxide (H(2)O(2)) levels, leading to a variety of physiological responses in plants. Catalase, which is involved in the degradation of H(2)O(2) into water and oxygen, is the major H(2)O(2)-scavenging enzyme in all aerobic organisms. A close interaction exists between intracellular H(2)O(2) and cytosolic calcium in response to biotic and abiotic stresses. Studies indicate that an increase in cytosolic calcium boosts the generation of H(2)O(2). Here we report that calmodulin (CaM), a ubiquitous calcium-binding protein, binds to and activates some plant catalases in the presence of calcium, but calcium/CaM does not have any effect on bacterial, fungal, bovine, or human catalase. These results document that calcium/CaM can down-regulate H(2)O(2) levels in plants by stimulating the catalytic activity of plant catalase. Furthermore, these results provide evidence indicating that calcium has dual functions in regulating H(2)O(2) homeostasis, which in turn influences redox signaling in response to environmental signals in plants.

NASA Discipline Plant Biology↗

An early ethylene up-regulated gene encoding a calmodulin-binding protein involved in plant senescence and death

35S-Labeled calmodulin (CaM) was used to screen a tobacco anther cDNA library. A positive clone (NtER1) with high homology to an early ethylene-up-regulated gene (ER66) in tomato, and an Arabidopsis homolog was isolated and characterized. Based on the helical wheel projection, a 25-mer peptide corresponding to the predicted CaM-binding region of NtER1 (amino acids 796-820) was synthesized. The gel-mobility shift assay showed that the peptide formed a stable complex with CaM only in the presence of Ca(2+). CaM binds to NtER1 with high affinity (K(d) approximately 12 nm) in a calcium-dependent manner. Tobacco flowers at different stages of development were treated with ethylene or with 1-methylcyclopropene for 2 h before treating with ethylene. Northern analysis showed that the NtER1 was rapidly induced after 15 min of exposure to ethylene. However, the 2-h 1-methylcyclopropene treatment totally blocked NtER1 expression in flowers at all stages of development, suggesting that NtER1 is an early ethylene-up-regulated gene. The senescing leaves and petals had significantly increased NtER1 induction as compared with young leaves and petals, implying that NtER1 is developmentally regulated and acts as a trigger for senescence and death. This is the first documented evidence for the involvement of Ca(2+)/CaM-mediated signaling in ethylene action.

Non-NASA Center↗

Arabidopsis chloroplast chaperonin 10 is a calmodulin-binding protein

Calcium regulates diverse cellular activities in plants through the action of calmodulin (CaM). By using (35)S-labeled CaM to screen an Arabidopsis seedling cDNA expression library, a cDNA designated as AtCh-CPN10 (Arabidopsis thaliana chloroplast chaperonin 10) was cloned. Chloroplast CPN10, a nuclear-encoded protein, is a functional homolog of E. coli GroES. It is believed that CPN60 and CPN10 are involved in the assembly of Rubisco, a key enzyme involved in the photosynthetic pathway. Northern analysis revealed that AtCh-CPN10 is highly expressed in green tissues. The recombinant AtCh-CPN10 binds to CaM in a calcium-dependent manner. Deletion mutants revealed that there is only one CaM-binding site in the last 31 amino acids of the AtCh-CPN10 at the C-terminal end. The CaM-binding region in AtCh-CPN10 has higher homology to other chloroplast CPN10s in comparison to GroES and mitochondrial CPN10s, suggesting that CaM may only bind to chloroplast CPN10s. Furthermore, the results also suggest that the calcium/CaM messenger system is involved in regulating Rubisco assembly in the chloroplast, thereby influencing photosynthesis. Copyright 2000 Academic Press.

NASA Discipline Plant Biology↗

Molecular and biochemical evidence for the involvement of calcium/calmodulin in auxin action

The use of (35)S-labeled calmodulin (CaM) to screen a corn root cDNA expression library has led to the isolation of a CaM-binding protein, encoded by a cDNA with sequence similarity to small auxin up RNAs (SAURs), a class of early auxin-responsive genes. The cDNA designated as ZmSAUR1 (Zea mays SAURs) was expressed in Escherichia coli, and the recombinant protein was purified by CaM affinity chromatography. The CaM binding assay revealed that the recombinant protein binds to CaM in a calcium-dependent manner. Deletion analysis revealed that the CaM binding site was located at the NH(2)-terminal domain. A synthetic peptide of amino acids 20-45, corresponding to the potential CaM binding region, was used for calcium-dependent mobility shift assays. The synthetic peptide formed a stable complex with CaM only in the presence of calcium. The CaM affinity assay indicated that ZmSAUR1 binds to CaM with high affinity (K(d) approximately 15 nM) in a calcium-dependent manner. Comparison of the NH(2)-terminal portions of all of the characterized SAURs revealed that they all contain a stretch of the basic alpha-amphiphilic helix similar to the CaM binding region of ZmSAUR1. CaM binds to the two synthetic peptides from the NH(2)-terminal regions of Arabidopsis SAUR-AC1 and soybean 10A5, suggesting that this is a general phenomenon for all SAURs. Northern analysis was carried out using the total RNA isolated from auxin-treated corn coleoptile segments. ZmSAUR1 gene expression began within 10 min, increased rapidly between 10 and 60 min, and peaked around 60 min after 10 microM alpha-naphthaleneacetic acid treatment. These results indicate that ZmSAUR1 is an early auxin-responsive gene. The CaM antagonist N-(6-aminohexyl)5-chloro-1-naphthalenesulfonamide hydrochloride inhibited the auxin-induced cell elongation but not the auxin-induced expression of ZmSAUR1. This suggests that calcium/CaM do not regulate ZmSAUR1 at the transcriptional level. CaM binding to ZmSAUR1 in a calcium-dependent manner suggests that calcium/CaM regulate ZmSAUR1 at the post-translational level. Our data provide the first direct evidence for the involvement of calcium/CaM-mediated signaling in auxin-mediated signal transduction.

NASA Program Fundamental Space Biology↗

Space Radiation Absorbed Dose Distribution in a Human Phantom Torso

The flight of a human phantom torso with head that containing active dosimeters at 5 organ sites and 1400 TLDs distributed in 34 1" thick sections is described. Experimental dose rates and quality factors are compared with calculations for shielding distributions at the sites using the Computerized Anatomical Male (CAM) model. The measurements were complemented with those obtained from other instruments. These results have provided the most comprehensive data set to map the dose distribution inside a human and to assess the accuracy of radiation transport models and astronaut radiation risk.

Badhwar, G. D.↗

Developmental regulation of the gene for chimeric calcium/calmodulin-dependent protein kinase in anthers

Chimeric Ca(2+)/calmodulin-dependent protein kinase (CCaMK) was cloned from developing anthers of lily (Lilium longiflorum Thumb. cv. Nellie White) and tobacco (Nicotiana tabacum L. cv. Xanthi). Previous biochemical characterization and structure/function studies had revealed that CCaMK has dual modes of regulation by Ca(2+) and Ca(2+)/calmodulin. The unique structural features of CCaMK include a catalytic domain, a calmodulin-binding domain, and a neural visinin-like Ca(2+)-binding domain. The existence of these three features in a single polypeptide distinguishes it from other kinases. Western analysis revealed that CCaMK is expressed in a stage-specific manner in developing anthers. Expression of CCaMK was first detected in pollen mother cells and continued to increase, reaching a peak around the tetrad stage of meiosis. Following microsporogenesis, CCaMK expression rapidly decreased and at later stages of microspore development, no expression was detected. A tobacco genomic clone of CCaMK was isolated and transgenic tobacco plants were produced carrying the CCaMK promoter fused to the beta-glucuronidase reporter gene. Both CCaMK mRNA and protein were detected in the pollen sac and their localizations were restricted to the pollen mother cells and tapetal cells. Consistent results showing a stage-specific expression pattern were obtained by beta-glucuronidase analysis, in-situ hybridization and immunolocalization. The stage- and tissue-specific appearance of CCaMK in anthers suggests that it could play a role in sensing transient changes in free Ca(2+) concentration in target cells, thereby controlling developmental events in the anther.

NASA Discipline Plant Biology↗

Theoretical and experimental tests of a chromosomal fingerprint for densely ionizing radiation based on F ratios calculated from stable and unstable chromosome aberrations

In the present study, F ratios for both stable chromosome aberrations, i.e. ratios of translocations to pericentric inversions, and unstable aberrations, i.e. dicentrics and centric rings, were measured using fluorescence in situ hybridization. F ratios for stable aberrations measured after exposure to low (2.89 Gy 60Co gamma rays) and high-LET (0.25 Gy 56Fe ions; 1.25 Gy 56Fe ions; 3.0 Gy 12C ions) radiation were 6.5 +/- 1.5, 4.7 +/- 1.6, 9.3 +/- 2.5 and 10.4 +/- 3.0, respectively. F ratios for unstable aberrations measured after low (2.89 Gy 60Co gamma rays) and high-LET (0.25 Gy 56Fe ions; 3.0 Gy 12C ions) radiations were 6.5 +/- 1.6, 6.3 +/- 2.3 and 11.1 +/- 3.7, respectively. No significant difference between the F ratios for low- and high-LET radiation was found. Further tests on the models for calculation of the F ratio proposed by Brenner and Sachs (Radiat. Res. 140, 134-142, 1994) showed that the F ratio may not be straightforward as a practical fingerprint for densely ionizing radiation.

NASA Center JSC↗

Experimental Studies on a Compact Storage Scheme for Wavelet-based Multiresolution Subregion Retrieval

Wavelet transforms, when combined with quantization and a suitable encoding, can be used to compress images effectively. In order to use them for image library systems, a compact storage scheme for quantized coefficient wavelet data must be developed with a support for fast subregion retrieval. We have designed such a scheme and in this paper we provide experimental studies to demonstrate that it achieves good image compression ratios, while providing a natural indexing mechanism that facilitates fast retrieval of portions of the image at various resolutions.

Poulakidas, A.↗

A seal test facility for the measurement of isotropic and anisotropic linear rotordynamic characteristics

A new seal test facility for measuring high-pressure seal rotor-dynamic characteristics has recently been made operational at Case Western Reserve University (CWRU). This work is being sponsored by the Electric Power Research Institute (EPRI). The fundamental concept embodied in this test apparatus is a double-spool-shaft spindle which permits independent control over the spin speed and the frequency of an adjustable circular vibration orbit for both forward and backward whirl. Also, the static eccentricity between the rotating and non-rotating test seal parts is easily adjustable to desired values. By accurately measuring both dynamic radial displacement and dynamic radial force signals, over a wide range of circular orbit frequency, one is able to solve for the full linear-anisotropic model's 12 coefficients rather than the 6 coefficients of the more restrictive isotropic linear model. Of course, one may also impose the isotropic assumption in reducing test data, thereby providing a valid qualification of which seal configurations are well represented by the isotropic model and which are not. In fact, as argued in reference (1), the requirement for maintaining a symmetric total system mass matrix means that the resulting isotropic model needs 5 coefficients and the anisotropic model needs 11 coefficients.

Adams, M. L.↗

An inverse problem design method for branched and unbranched axially symmetrical ducts

This paper concerns the potential flow design of axially symmetrical ducts of both circular and annular cross section with or without wall suction or blowing slots. The objective of the work was to develop a method by which such ducts could be designed with directly prescribed wall pressure variation. Previous axially symmetrical design methods applied only to circular cross sectional ducts and required that the pressure distribution be prescribed along the duct centerline and not along the duct wall. The present method uses an inverse problem approach which extends the method of Stanitz to the axially symmetrical case, and an approximation is used to account for the stagnation point in branched duct designs. Two examples of successful designs of diffusers with suction slots are presented.

Nelson, C. D.↗

The design and performance of axially symmetrical contoured wall diffusers employing suction boundary layer control

This paper presents a procedure for the design and the performance prediction of axially symmetrical contoured wall diffusers employing suction boundary layer control. An inverse problem approach was used in the potential flow design of the diffuser wall contours. The experimentally observed flow characteristics and the stability of flows within the diffuser are also described. Guidelines for the design of low suction (less than 10 percent of the inlet flow) and thus high effectiveness contoured wall diffusers are also provided based on the results of the experimental program.

Nelson, C. D., Jr.↗

Design and experimental performance of short curved wall diffusers with axial symmetry utilizing slot suction

The feasibility of designing short curved wall axially symmetrical subsonic diffusers utilizing suction through slots in the diffuser walls to prevent flow separation was investigated. A potential flow analysis was made, and a digital computer program was written for determining the diffuser wall contour for prescribed boundary conditions. The flow field included branch flow so that the suction slot geometry could be a part of the diffuser design. One bell shaped diffuser and three annular diffusers with area ratios of either 2.5:1 or 3:1 were designed, fabricated, and tested. Minimum suction requirements of metastable operation ranged from 6.3 percent to 12 percent when operating with inlet air velocities in the 1000 to 250 ft/sec (30 to 76 m/sec) range. For stable operation suction rates from 10 percent to 22 percent were required. In all cases the diffuser effectiveness was above 95 percent based on the conventional definition, and from 81 percent to 94 percent when the suction loss was accounted for. The exit velocity profiles were virtually flat with no more than + or - 9% variation over 95 percent of the exit area when operated with sufficient suction to prevent flow separation.

Yang, T.↗

An improved design method and experimental performance of two dimensional curved wall diffusers

A computer design program was developed to incorporate the suction slots in solving the potential flow equations with prescribed boundary conditions. Using the contour generated from this program two Griffith diffusers were tested having area ratios AR = 3 and 4. The inlet Reynolds number ranged from 600,000 to 7 million. It was found that the slot suction required for metastable operation depends on the sidewall suction applied. Values of slot suction of 8% of the inlet flow rate was required for AR = 4 with metastable condition, provided that enough sidewall suction was applied. For AR = 3, the values of slot suction was about 25% lower than those required for AR = 4. For nearly all unseparated test runs, the effectiveness was 100% and the exit flow was uniform. In addition to the Griffith diffusers, dump and cusp diffusers of comparable area ratios were built and tested. The results obtained from these diffusers were compared with those of the Griffith diffusers. Flow separation occurred in all test runs with the dump and cusp diffusers.

Yang, T.↗