Differences in Gene Expression of Pear Selections Showing Leaf Curling or Leaf Reddening Symptoms Due to Pear Decline Phytoplasma.

While host gene expression has been related to symptoms associated with different phytoplasma diseases, it is unknown why some phytoplasmas are associated with different symptoms in genotypes of the same plant species. Pear tree selections showed symptoms of either leaf reddening (selection 8824-1) or leaf curling (selection 9328-1) associated with pear decline (PD) phytoplasma presence. PD populations were similar in leaves and shoots of the two selections, but in the roots, populations were significantly lower in selection 8824-1 than in 9328-1, indicating greater resistance. For host carbohydrate metabolism gene expression in PD-infected tissues, significant up-regulation in selection 8824-1 was observed for a sucrose synthase gene in leaves and an acid invertase gene in leaves and roots. These features have been associated with localized higher sugar levels in phytoplasma-infected tissues, and thus may be related to leaf reddening. For host stress/defense response gene expression in PD-infected tissues, significant up-regulation of a phenylalanine ammonia lyase gene was observed in PD-infected shoots of both selections; however, up-regulation of alcohol dehydrogenase gene in shoots, a chitinase gene in all tissues and a phloem protein 2 gene in roots was only observed for selection 8824-1. These changes indicate greater triggered innate immunity in roots associated with lower PD populations and leaf reddening. Leaf reddening may be related to changes in gene expression associated with increased sugar levels in leaves and stronger immune responses in several tissues, while leaf curling may be due to water stress resulting from dysfunctional root associated with higher PD populations in the roots.

Increase of Albinistic Hosts Caused by Gut Parasites Promotes Self-Transmission.

Paranosema locustae is a gut parasite that has been applied widely in the control of grasshoppers in many parts of the world. Usually, P. locustae is transmitted horizontally via passive modes under natural conditions but in the current study, a positive transmission strategy of P. locustae was demonstrated. First, infection by P. locustae resulted in the cuticula of infected Locusta migratoria nymphs to become lighter in color: normally only a small proportion of locusts are pale with most either being partly or mostly black; but locusts infected with P. locustae became pale. And it was found that the change to pale occurred even among uninfected black and partly black nymphs reared with infected locusts. The eumelanin of the thorax and abdomen of infected individuals decreased significantly, as did the level of dopamine. In addition, there was a decrease in phenol oxidase activity and the expression of henna and pale, which are involved in the synthesis of cuticle melanin, decreased. What is the ecological significance of this increase in light-colored hosts caused by P. locustae? We discovered that light-colored locusts were more susceptible to the microsporidian pathogen than dark-colored individuals were, because of their weaker melanization. Phenol oxidase activity in pale locusts was lower than that of black locusts, but the serpin expression level of pale locusts was higher than that of black individuals. When examined for infection, it was found that initially uninfected nymphs had picked up P. locustae infections indicating that infections are readily passed from one pale locust to another. The infection rate of healthy locusts reared with light-colored locusts infected with P. locustae was 100% which was more than with black-colored ones. The increase in albinistic locusts clearly promoted the prevalence of P. locustae in the total population. In conclusion, these results elucidated a new strategy of positive self-transmission in P. locustae. Importance: Mother Nature always grants wisdom to her creatures and feeds them carefully. This wisdom is particularly apparent in the relationships between two interacting species. In this study, our team focused on the interaction between L. migratoria and P. locustae. In a previous study, it was found that L. migratoria isolate infected individuals, reducing avoiding the spread of P. locustae, in a previous study. The solitary, pale individuals infected by P. locustae were left behind as locust groups marched ahead, leading to a kind of behavioral immunity in the insects. Here, we reported that P. locustae promotes pigmentation loss in L. migratoria, causing a larger proportion of light-colored individuals, and these lighter individuals which possessed weaker immunity against pathogens. This strategy is advantageous to P. locustae, as it promotes its propagation and spread. These extraordinary abilities of L. migratoria and P. locustae have accumulated over millennia of years of interaction.

Comparative performance of modern digital mammography systems in a large breast screening program.

PURPOSE: To compare physical measures pertaining to image quality among digital mammography systems utilized in a large breast screening program. To examine qualitatively differences in these measures and differences in clinical cancer detection rates between CR and DR among sites within that program. METHODS: As part of the routine quality assurance program for screening, field measurements are made of several variables considered to correlate with the diagnostic quality of medical images including: modulation transfer function, noise equivalent quanta, d' (an index of lesion detectability) and air kerma to allow estimation of mean glandular dose. In addition, images of the mammography accreditation phantom are evaluated. RESULTS: It was found that overall there were marked differences between the performance measures of DR and CR mammography systems. In particular, the modulation transfer functions obtained with the DR systems were found to be higher, even for larger detector element sizes. Similarly, the noise equivalent quanta, d', and the phantom scores were higher, while the failure rates associated with low signal-to-noise ratio and high dose were lower with DR. These results were consistent with previous findings in the authors' program that the breast cancer detection rates at sites employing CR technology were, on average, 30.6% lower than those that used DR mammography. CONCLUSIONS: While the clinical study was not large enough to allow a statistically powered system-by-system assessment of cancer detection accuracy, the physical measures expressing spatial resolution, and signal-to-noise ratio are consistent with the published finding that sites employing CR systems had lower cancer detection rates than those using DR systems for screening mammography.

Biological activity of the tzs gene of nopaline Agrobacterium tumefaciens GV3101 in plant regeneration and genetic transformation.

Agrobacterium tumefaciens has been widely used in plant genetic transformation. Hormone-encoding genes residing in the T-DNA region have been removed, resulting in disarmed Agrobacterium strains that are used in various transformation experiments. Nopaline Agrobacterium strains, however, carry another hormone gene, trans-zeatin synthesizing (tzs), that codes for trans-zeatin in the virulence region of the tumor-inducing plasmids. We investigated the activity and function of the tzs gene of a nopaline Agrobacterium sp. strain GV3101 in plant in vitro regeneration. Leaf explants of tobacco and Nicotiana benthamiana co-cultured with strain GV3101 exhibited active shoot regeneration in media without added plant growth regulators. On medium without plant growth regulators, transgenic shoots were also induced from explants co-cultured with GV3101 containing a binary vector. Enzyme-linked immunosorbent assay showed that cell-free extracts of Agrobacterium sp. strain GV3101 culture contained the trans-zeatin at 860 ng/liter. Polymerase chain reaction using tzs-specific primers showed that the tzs gene was present in strain GV3101 but not in other Agrobacterium strains. The study showed that the tzs gene in GV3101 was actively expressed, and that trans-zeatin produced in the Agrobacterium strain can promote plant shoot regeneration.

Measured and calculated K-fluorescence effects on the MTF of an amorphous-selenium based CCD x-ray detector.

PURPOSE: Theoretical reasoning suggests that direct conversion digital x-ray detectors based upon photoconductive amorphous-selenium (a-Se) could attain very high values of the MTF (modulation transfer function) at spatial frequencies well beyond 20 cycles mm(-1). One of the fundamental factors affecting resolution loss, particularly at x-ray energies just above the K-edge of selenium (12.66 keV), is the K-fluorescence reabsorption mechanism, wherein energy can be deposited in the detector at locations laterally displaced from the initial x-ray interaction site. This paper compares measured MTF changes above and below the Se K-edge of a CCD based a-Se x-ray detector with theoretical expectations. METHODS: A prototype 25 µm sampling pitch (Nyquist frequency = 20 cycles mm(-1), 200 µm thick a-Se layer based x-ray detector, utilizing a specialized CCD readout device (200 × 400 area array), was used to make edge images with monochromatic x-rays above and below the K-edge of Se. A vacuum double crystal monochromator, exposed to polychromatic x-rays from a synchrotron, formed the monochromatic x-ray source. The monochromaticity of the x-rays was 99% or better. The presampling MTF was determined using the slanted edge method. The theory modeling the MTF performance of the detector includes the basic x-ray interaction physics in the a-Se layer as well as effects related to the operation of the CCD and charge trapping at a blocking layer present at the CCD/a-Se interface. RESULTS: The MTF performance of the prototype a-Se CCD was reduced from the theoretical value prescribed by the basic Se x-ray interaction physics, principally by the presence of a blocking layer. Nevertheless, the K-fluorescence reduction in the MTF was observed, approximately as predicted by theory. For the CCD prototype detector, at five cycles mm(-1), there was a 14% reduction of the MTF, from a value of 0.7 below the K-edge of Se, to 0.6 just above the K-edge. CONCLUSIONS: The MTF of an a-Se x-ray detector has been measured using monochromatic x-rays above and below the K-edge of selenium. The MTF is poorer above the K-edge by an amount consistent with theoretical expectations.

Pi-interaction tuning of the active site properties of metalloproteins.

The influence of pi-interactions with a His ligand have been investigated in a family of copper-containing redox metalloproteins. The Met16Phe and Met16Trp pseudoazurin, and Leu12Phe spinach and Leu14Phe Phormidium laminosum plastocyanin variants possess active-site pi-contacts between the introduced residue and His81 and His87/92 respectively. The striking overlap of the side chain of Phe16 in the Met16Phe variant and that of Met16 in wild type pseudoazurin identifies that this position provides an important second coordination sphere interaction in both cases. His-ligand protonation and dissociation from Cu(I) occurs in the wild type proteins resulting in diminished redox activity, providing a [H(+)]-driven switch for regulating electron transfer. The introduced pi-interaction has opposing effects on the pKa for the His ligand in pseudoazurin and plastocyanin due to subtle differences in the pi-contact, stabilizing the coordinated form of pseudoazurin whereas in plastocyanin protonation and dissociation is favored. Replacement of Pro36, a residue that has been suggested to facilitate structural changes upon His ligand protonation, with a Gly, has little effect on the pKa of His87 in spinach plastocyanin. The mutations at Met16 have a significant influence on the reduction potential of pseudoazurin. Electron self-exchange is enhanced, whereas association with the physiological partner, nitrite reductase, is only affected by the Met16Phe mutation, but kcat is halved in both the Met16Phe and Met16Trp variants. Protonation of the His ligand is the feature most affected by the introduction of a pi-interaction.

The dependence of the modulation transfer function on the blocking layer thickness in amorphous selenium x-ray detectors.

Blocking layers are used to reduce leakage current in amorphous selenium detectors. The effect of the thickness of the blocking layer on the presampling modulation transfer function (MTF) and on dark current was experimentally determined in prototype single-line CCD-based amorphous selenium (a-Se) x-ray detectors. The sampling pitch of the detectors evaluated was 25 microm and the blocking layer thicknesses varied from 1 to 51 microm. The blocking layers resided on the signal collection electrodes which, in this configuration, were used to collect electrons. The combined thickness of the blocking layer and a-Se bulk in each detector was approximately 200 microm. As expected, the dark current increased monotonically as the thickness of the blocking layer was decreased. It was found that if the blocking layer thickness was small compared to the sampling pitch, it caused a negligible reduction in MTF. However, the MTF was observed to decrease dramatically at spatial frequencies near the Nyquist frequency as the blocking layer thickness approached or exceeded the electrode sampling pitch. This observed reduction in MTF is shown to be consistent with predictions of an electrostatic model wherein the image charge from the a-Se is trapped at a characteristic depth within the blocking layer, generally near the interface between the blocking layer and the a-Se bulk.

Lag and ghosting in a clinical flat-panel selenium digital mammography system.

We present measurements of lag and ghosting in a FDA-approved digital mammography system that uses a dielectric/selenium based detector structure. Lag is the carryover of signal from a previous image, whereas ghosting is the reduction of sensitivity caused by previous exposure history of the detector. Data from six selenium units were acquired. For the type of selenium detector tested, and under typical clinical usage conditions, the lag was as high as 0.15% of source signal and the ghosting could be as high as 15%. The amount of lag and ghosting varied from unit to unit. Results were compared with data acquired on a phosphor-based full-field digital mammography system. Modifications in the technology of the selenium detectors appear to have resulted in a marked decrease in both lag and ghosting effects in more recent systems.

The parsley plastocyanin-turnip cytochrome f complex: a structurally distorted but kinetically functional acidic patch.

In general, inter-protein electron transfer proceeds via the formation of transient complexes. The initial stage of the interaction between plastocyanin (PCu) and cytochrome f (cyt f ) from plants is mediated by complementary electrostatics. Given the diffuse nature of its acidic patch, parsley PCu is an atypical example of a plant PCu. The interaction of this PCu with turnip cyt f was investigated by stopped-flow kinetics, NMR spectroscopy and protein-docking simulations. We show that, despite the altered acidic patch, parsley PCu is as efficient as spinach PCu in accepting electrons from cyt f, over the physiological range of ionic strength. At high ionic strength, the rate constant for the reaction of cyt f with parsley PCu is twice that of the spinach protein. This difference in reactivity is attributed to variations in the hydrophobic patch of parsley PCu. The results of NMR studies and protein-docking simulations indicate that parsley PCu and its spinach analogue adopt different orientations in their complexes with cyt f.

Risks to the aquatic ecosystem from the application of Metarhizium anisopliae for locust control in Australia.

Laboratory tests of Metarhizium anisopliae var acridum Driver & Milner, at a dose of 1.3 x 10(6) conidia ml-1, had no adverse effects on nymphs of mayfly, Ulmerophlebia sp or 8-week-old fry of the rainbow fish, Melanotaenia duboulayi Castelnau. This dose was toxic to the cladoceran, Ceriodaphnia dubia Richard, causing 100% mortality in 48 h. When this test was repeated at doses of up to 6.7 x 10(3) conidia ml-1, there was only 5% mortality after 192 h. Spraying of artificial water sources with a very high dose of the fungus as an aqueous spray resulted in 80-130 conidia ml-1 at 15 cm depth in the first 24 h after spraying. The conidia rapidly settled out and were absent from the top 15 cm layer of water after about 50 h. A similar experiment using the oil formulation as used in field control resulted in a 2- to 20-fold lower level of conidia in the water. Finally, sampling actual water sources in spray areas revealed a very low level of contamination of the water, with a maximum mean level of 29 conidia ml-1 in the first 24 h after treatment. Thus the level of conidia likely to enter water during control campaigns is a small fraction of that required to kill cladocerans, the only sensitive non-target organism tested. It is concluded that the biopesticide is very unlikely to pose any hazard to aquatic organisms.

The clinical significance of anterior horn meniscal tears diagnosed on magnetic resonance images.

We assessed the accuracy of magnetic resonance imaging in detecting clinically significant lesions of the anterior horn of the meniscus by reviewing 947 consecutive knee magnetic resonance imaging reports. Of these, 76 (8%) indicated a tear of the anterior horn of the medial or lateral meniscus. Thirty-one of these 76 patients underwent a subsequent arthroscopic examination, and their operative reports were reviewed. The 45 patients who were not examined arthroscopically were contacted and interviewed for clinical follow-up. Among the 31 patients who underwent arthroscopic examination, 8 anterior horn tears were noted in the predicted area (26% true-positive results), 23 patients had intact anterior horns (74% false-positive results), and 18 had normal intact menisci in all zones. Of the 45 patients who did not undergo arthroscopic surgery, 6 had isolated anterior horn tears reported on magnetic resonance imaging, and 5 of the 6 were asymptomatic at follow-up. The other 39 patients had multiple pathologic conditions noted on the magnetic resonance imaging report and continued to report knee pain at the follow-up interview. Increased signal intensity at the anterior horn of the meniscus seen on magnetic resonance imaging commonly does not represent a clinically significant lesion. We recommend correlation with the physical examination when interpreting this "positive" finding on knee magnetic resonance imaging examinations.