Best Practice Guidelines for Pre-Launch Characterization and Calibration of Instruments for Passive Optical Remote Sensing.

The pre-launch characterization and calibration of remote sensing instruments should be planned and carried out in conjunction with their design and development to meet the mission requirements. The onboard calibrators such as blackbodies and the sensors such as spectral radiometers should be characterized and calibrated using SI traceable standards. In the case of earth remote sensing, this allows inter-comparison and intercalibration of different sensors in space to create global time series of climate records of high accuracy where some inevitable data gaps can be easily bridged. The recommended best practice guidelines for this pre-launch effort is presented based on experience gained at National Institute of Standards and Technology (NIST), National Aeronautics and Space Administration (NASA) and National Oceanic and Atmospheric Administration (NOAA) programs over the past two decades. The currently available radiometric standards and calibration facilities at NIST serving the remote sensing community are described. Examples of best practice calibrations and intercomparisons to build SI (international System of Units) traceable uncertainty budget in the instrumentation used for preflight satellite sensor calibration and validation are presented.

The transcriptional landscape of polyploid wheat.

The coordinated expression of highly related homoeologous genes in polyploid species underlies the phenotypes of many of the world's major crops. Here we combine extensive gene expression datasets to produce a comprehensive, genome-wide analysis of homoeolog expression patterns in hexaploid bread wheat. Bias in homoeolog expression varies between tissues, with ~30% of wheat homoeologs showing nonbalanced expression. We found expression asymmetries along wheat chromosomes, with homoeologs showing the largest inter-tissue, inter-cultivar, and coding sequence variation, most often located in high-recombination distal ends of chromosomes. These transcriptionally dynamic genes potentially represent the first steps toward neo- or subfunctionalization of wheat homoeologs. Coexpression networks reveal extensive coordination of homoeologs throughout development and, alongside a detailed expression atlas, provide a framework to target candidate genes underpinning agronomic traits in wheat.

Genetic transformation of peanut (Arachis hypogaea L.) using cotyledonary node as explant and a promoterless gus::nptII fusion gene based vector.

We have generated putative promoter tagged transgenic lines in Arachis hypogaea cv JL-24 using cotyledonary node (CN) as an explant and a promoterless gus::nptII bifunctional fusion gene mediated by Agrobacterium transformation. MS medium fortified with 6-benzylaminopurine (BAP) at 4mg/l in combination with 0.1 mg/l alpha -napthaleneacetic acid (NAA) was the most effective out of the various BAP and NAA combinations tested in multiple shoot bud formation. Parameters enhancing genetic transformation viz. seedling age, Agrobacterium genetic background and co-cultivation periods were studied by using the binary vector p35SGUSINT. Genetic transformation with CN explants from 6-day-old seedlings co-cultivated with Agrobacterium GV2260 strain for 3 days resulted in high kanamycin resistant shoot induction percentage (45%); approximately 31% transformation frequency was achieved with p35S GUSINT in beta-glucuronidase (GUS) assays. Among the in vivo GUS fusions studied with promoterless gus::nptII construct, GUS-positive sectors occupied 38% of the total transient GUS percentage. We have generated over 141 putative T 0 plants by using the promoterless construct and transferred them to the field. Among these, 82 plants survived well in the green house and 5 plants corresponding to 3.54% showed stable integration of the fusion gene as evidenced by GUS, polymerase chain reaction (PCR) and Southern blot analyses. Twenty-four plants were positive for GUS showing either tissue-specific expression or blue spots in at least one plant part. The progeny of 15 T 0 plants indicated Mendelian inheritance pattern of segregation for single-copy integration. The tissue-specific GUS expression patterns were more or less similar in both T 0 and corresponding T 1 progeny plants. We present the differential patterns of GUS expression identified in the putative promoter-tagged transgenic lines in the present communication.

Modified binary plant transformation vectors with the wild-type gene encoding NPTII.

The defective gene encoding neomycin phosphotransferase (NPTII) present in the binary plasmid vector, pBin19, was replaced with the wild-type (wt) gene. Plasmid vectors analogous to pBin19, pBI121 and pBI101 were constructed carrying the gene encoding the wt NPTII enzyme activity.

A bifunctional fusion between beta-glucuronidase and neomycin phosphotransferase: a broad-spectrum marker enzyme for plants.

We have used an in vivo selection approach to isolate a gene encoding a bifunctional fusion peptide between Escherichia coli beta-glucuronidase (GUS) and neomycin phosphotransferase II (NPT-II) from transposon Tn5 in the NH2-GUS::NPT-II-COOH configuration. The fused gene is predicted to encode a fusion peptide 885 amino acids long, and was shown in E. coli to synthesize a 97-kDa GUS+ NPT-II+ gene product. Gel-filtration chromatography suggested that, while the native GUS may be active as a dimer and NPT-II as a monomer, the elution profile of the fusion protein is consistent with that of a trimer. The fusion marker has been produced and defined in transgenic Nicotiana tabacum plants, where both the chimeric gene and the gene product were stable. The bifunctional gene enabled direct KmR selection at the callus stage and enzymatic or histochemical assessment of the steady-state production of GUS activity in regenerated plants. In addition to allowing structure-function determination for the GUS and NPT-II domains of the fusion peptide, the gus::npt-II gene simplifies vector constructs where both marker domains are desired.

Development of an efficient Agrobacterium-mediated transformation system for Brassica carinata.

Shoot organogenesis and plant regeneration were readily achieved from cotyledonary petioles and hypocotyls of Brassica carinata. These explants were used for Agrobacterium-mediated transformation. A construct containing the selectable marker genes, neomycin phosphotransferase II, phosphinothricin acetyl transferase and the reporter gene ß-glucuronidase, under the control of a tandem 35S promoter, was used for transformation. Although transformation was achieved with both cotyledonary petioles and hypocotyls, cotyledonary petioles responded best, with 30-50% of the explants producing GUS-positive shoots after selection on 25 mg/l kanamycin. Direct selection on L-phosphinothricin also produced resistant shoots but at a lower frequency (1-2%).

The promoter of aBrassica napus polygalacturonase gene directs pollen expression ofß-glucuronidase in transgenicBrassica plants.

A 647-bp 5'-flanking fragment obtained from genomic clone Sta 44G(2) belonging to a family of polygalacturonase genes expressed inBrassica napus pollen was fused to theß-glucuronidase (GUS) marker gene. This fusion construct was introduced intoB. napus plants viaAgrobacterium tumefaciens transformation. Analysis of the transgenicB. napus plants revealed that this promoter fragment is sufficient to direct GUS expression specifically in the anther and that GUS activity increases in pollen during maturation.

Optimally Toothed Apertures for Reduced Diffraction.

We model diffraction errors found when using toothed apertures [L. P. Boivin, Reduction of diffraction errors in radiometry by means of toothed apertures, Appl. Opt. 17, 3323-3328 (1978)]. Using toothed (cf. circular) apertures minimizes diffraction by inducing destructive interference within the diffracted signal. Since diffraction effects can be quite complicated, their over-all reduction may help limit uncertainties in, say calibrations. Our analysis yields three principles to guide design of nonlimiting (baffle) apertures which minimize diffrac tion. We performed detailed diffraction calculations within scalar (Kirchoff) diffraction theory, using parallel-computing resources at the National Institute of Standards and Technology.

Development of a Bolometer Detector System for the NIST High Accuracy Infrared Spectrophotometer.

A bolometer detector system was developed for the high accuracy infrared spectrophotometer at the National Institute of Standards and Technology to provide maximum sensitivity, spatial uniformity, and linearity of response covering the entire infrared spectral range. The spatial response variation was measured to be within 0.1 %. The linearity of the detector output was measured over three decades of input power. After applying a simple correction procedure, the detector output was found to deviate less than 0.2 % from linear behavior over this range. The noise equivalent power (NEP) of the bolometer system was 6 × 10-12 [Formula: see text] at the frequency of 80 Hz. The detector output 3 dB roll-off frequency was 200 Hz. The detector output was stable to within ± 0.05 % over a 15 min period. These results demonstrate that the bolometer detector system will serve as an excellent detector for the high accuracy infrared spectrophotometer.

Erratum: Statistical Interpretation of Key Comparison Reference Value and Degrees of Equivalence.

[This corrects the article on p. 439 in vol. 108.].

Cryogenic Blackbody Calibrations at the National Institute of Standards and Technology Low Background Infrared Calibration Facility.

The Low Background Infrared Calibration Facility (LBIR) at the National Institute of Standards and Technology has been in operation for calibration measurements of the radiant power emitted from infrared radiation (IR) sources, such as cryogenic blackbodies, for more than 2 years. The IR sources are sent to NIST by customers from industry, government, and university laboratories. An absolute cryogenic radiometer is used as the standard detector to measure the total radiant power at its aperture. The low background is provided by a closed cycle helium refrigeration system that maintains the inner parts of the calibration chamber at 20 K. The radiance temperature of the blackbody is deduced from the measured power and compared with the blackbody temperature sensor data. The calibration procedures and data analysis are illustrated using the measurements of a typical blackbody.

Statistical Interpretation of Key Comparison Reference Value and Degrees of Equivalence.

Key comparisons carried out by the Consultative Committees (CCs) of the International Committee of Weights and Measures (CIPM) or the Bureau International des Poids et Mesures (BIPM) are referred to as CIPM key comparisons. The outputs of a statistical analysis of the data from a CIPM key comparison are the key comparison reference value, the degrees of equivalence, and their associated uncertainties. The BIPM publications do not discuss statistical interpretation of these outputs. We discuss their interpretation under the following three statistical models: nonexistent laboratory-effects model, random laboratory-effects model, and systematic laboratory-effects model.

Excretion studies of nitrofurantoin and nitrofurantoin with deglycyrrhizinated liquorice.

Nitrofurantoin and nitrofurantoin with liquorice were given to healthy volunteers and patients suffering from urinary tract infections. The excretion rates of the drug, colony counts and side effects were studied in patients and excretion rates in the volunteers. The excretion rates of the drug were significantly higher in patients receiving the drug with liquorice and also side effects were minimal. There was no significant difference in the excretion rates of the drug with addition of liquorice in healthy volunteers.

Experimental measurements and noise analysis of a cryogenic radiometer.

A cryogenic radiometer device, intended for use as part of an electrical-substitution radiometer, was measured at low temperature. The device consists of a receiver cavity mechanically and thermally connected to a temperature-controlled stage through a thin-walled polyimide tube which serves as a weak thermal link. With the temperature difference between the receiver and the stage measured in millikelvin and the electrical power measured in picowatts, the measured responsivity was 4700 K/mW and the measured thermal time constant was 14 s at a stage temperature of 1.885 K. Noise analysis in terms of Noise Equivalent Power (NEP) was used to quantify the various fundamental and technical noise contributions, including phonon noise and Johnson-Nyquist noise. The noise analysis clarifies the path toward a cryogenic radiometer with a noise floor limited by fundamental phonon noise, where the magnitude of the phonon NEP is 6.5 fW/√Hz for the measured experimental parameters.

Overexpression of human N-myristoyltransferase utilizing a T7 polymerase gene expression system.

Myristoyl CoA:protein N-myristoyltransferase catalyzes the addition of myristate to the amino-terminal glycine residue of a number of eukaryotic proteins. The gene encoding human N-myristoyltransferase (hNMT) was cloned into the overexpression vector pT7-7 which utilizes the T7 RNA polymerase gene expression system. The hNMT enzyme was purified to near homogeneity with more than 95% recovery using a single-step purification method involving SP-Sepharose fast flow column chromatography. The specific activity of the purified NMT was 220 nmol/min/mg of protein in the presence of oncoprotein-derived peptide substrate pp60src. The hNMT exhibited an apparent molecular weight of 49 kDa on SDS-polyacrylamide gel electrophoresis. Antibodies to Escherichia coli-expressed hNMT specifically recognize hNMT from crude bacterial lysates. The over-expressed hNMT was homogeneous and showed enzyme activity.

Genomic organization of human myristoyl-CoA: protein N-myristoyltransferase-1.

Myristoylation is a biochemical modification of proteins in which the lipid myristate becomes covalently bound to various cellular, viral, and oncoproteins catalyzed by a monomeric enzyme myristoyl-CoA:protein N-myristoyltransferase (NMT). This modification is important for the biological activity of several proteins, especially the regulation of several oncoproteins involved in various types of cancers. Complementary DNA encoding human NMT-1 (hNMT-1) has been previously reported; however, the genomic organization of hNMT-1 has not been available. Attempts to amplify genomic fragments corresponding to hNMT-1 cDNA sequence yielded only one fragment. We have searched databases using both the cDNA and sequence of one of the intron sequence and this identified a human BAC clone sequence from chromosome 17. Alignment of hNMT-1 cDNA coding information on human chromosome 17 resulted in the complete structural identity of 23,960 bp of the hNMT-1 gene. The hNMT-1 gene is composed of 11 exons and 10 introns with consensus GT/AG boundaries. Finally, we show that 140 bp from the 5' end of recently reported full-length cDNA of hNMT-1 was not part of this genomic region raising the possibility for posttranscriptional modification in generating larger transcripts likely by trans splicing. Further, the availability of this genomic sequence will assist in unraveling the molecular basis for several observed NMT isoforms.

Expression of human N-myristoyltransferase in Escherichia coli. Comparison with N-myristoyltransferases expressed in different tissues.

Myristoyl CoA:protein N-myristoyltransferase catalyzes the addition of myristate to the amino-terminal glycine residue of a number of eukaryotic proteins. Escherichia coli transformed with human NMT expression construct produced high levels of N-myristoyltransferase. Using the combination of ammonium sulfate precipitation, chromatography on SP-Sepharose fast flow and fast protein liquid chromatography on Mono-S, the enzyme was purified more than 100 fold with 40% yield. The hNMT fusion protein exhibited an apparent molecular weight of 53 kDa on SDS-polyacrylamide gel electrophoresis. Upon cleavage by the Enterokinase [(Asp)4-Lys], the hNMT exhibited an apparent molecular mass of 49 kDa without loss of catalytic activity. The hNMT activity could be greatly activated severalfold with the use of Tris, SDS, ethanol and acetonitrile. The catalytic activity of hNMT was potently inhibited in a concentration dependent manner by NIP71, a bovine brain NMT inhibitory protein with a half maximal inhibition of 31.0 nM. The E. coli expressed hNMT was homogeneous and showed enzyme activity.