Contrast medium injection protocols for coronary CT angiography: should contrast medium volumes be tailored to body weight or body surface area?

AIM: To compare the uniformity and image quality between contrast media injection protocols adjusted for patient body weight (BW) versus body surface area (BSA) during coronary computed tomography (CT) angiography (CCTA). MATERIALS AND METHODS: Consecutive patients (n=489) with suspected coronary artery disease were randomised prospectively to one of two CCTA protocols. In the BW protocol (n=245), patients received individualised iodine delivery rates (≤50 kg: 1 g/s; 51-60 kg: 1.2 g/s; 61-70 kg: 1.4 g/s; 71-80 kg: 1.6 g/s; 81-90 kg: 1.8 g/s; 91-100 kg: 2 g/s; >100 kg: 2.2 g/s). In the BSA protocol (n=244), patients received 9,600 mg iodine/m2 of contrast medium over 12 seconds. Attenuation and image noise were measured. Signal-to-noise ratio and contrast-to-noise ratio were calculated. Image quality was scored. Attenuation was assessed for correlation with BW and BSA using linear regression. RESULTS: There were no statistically significant differences in mean arterial attenuation (396.8±47.6 versus 395.8±42.2 HU, p=0.804; 95% confidence interval: -7 to 9), image noise (25.2±5.8 versus 25.5±5.4 HU; p=0.549), signal-to-noise ratio (16.7±4.4 versus 16.6±3.6; p=0.902), contrast-to-noise ratio (25.1±5.8 versus 25.8±7.4; p=0.258) or image quality scores (4.1±0.9 versus 4±0.9; p=0.770) between the BW and BSA protocols. There was no correlation between BW and aortic attenuation or between BSA and aortic attenuation (p=0.324 and 0.932, respectively). CONCLUSION: The average contrast media attenuation and image quality was comparable between BW-adjusted protocol and BSA-adjusted protocol.

A Global Perspective on the Population Structure and Reproductive System of Phyllosticta citricarpa.

The citrus pathogen Phyllosticta citricarpa was first described 117 years ago in Australia; subsequently, from the summer rainfall citrus-growing regions in China, Africa, and South America; and, recently, the United States. Limited information is available on the pathogen's population structure, mode of reproduction, and introduction pathways, which were investigated by genotyping 383 isolates representing 12 populations from South Africa, the United States, Australia, China, and Brazil. Populations were genotyped using seven published and eight newly developed polymorphic simple-sequence repeat markers. The Chinese and Australian populations had the highest genetic diversities, whereas populations from Brazil, the United States, and South Africa exhibited characteristics of founder populations. The U.S. population was clonal. Based on principal coordinate and minimum spanning network analyses, the Chinese populations were distinct from the other populations. Population differentiation and clustering analyses revealed high connectivity and possibly linked introduction pathways between South Africa, Australia, and Brazil. With the exception of the clonal U.S. populations that only contained one mating type, all the other populations contained both mating types in a ratio that did not deviate significantly from 1:1. Although most populations exhibited sexual reproduction, linkage disequilibrium analyses indicated that asexual reproduction is important in the pathogen's life cycle.

Comparing Avocado, Swamp Bay, and Camphortree as Hosts of Raffaelea lauricola Using a Green Fluorescent Protein (GFP)-Labeled Strain of the Pathogen.

Raffaelea lauricola, a fungal symbiont of the ambrosia beetle Xyleborus glabratus, causes laurel wilt in members of the Lauraceae plant family. North American species in the family, such as avocado (Persea americana) and swamp bay (P. palustris), are particularly susceptible to laurel wilt, whereas the Asian camphortree (Cinnamomum camphora) is relatively tolerant. To determine whether susceptibility is related to pathogen colonization, a green fluorescent protein-labeled strain of R. lauricola was generated and used to inoculate avocado, swamp bay, and camphortree. Trees were harvested 3, 10, and 30 days after inoculation (DAI), and disease severity was rated on a 1-to-10 scale. By 30 DAI, avocado and swamp bay developed significantly more severe disease than camphortree (mean severities of 6.8 and 5.5 versus 1.6, P < 0.003). The extent of xylem colonization was recorded as the percentage of lumena that were colonized by the pathogen. More xylem was colonized in avocado than camphortree (0.9% versus 0.1%, P < 0.03) but colonization in swamp bay (0.4%) did not differ significantly from either host. Although there were significant correlations between xylem colonization and laurel wilt severity in avocado (r = 0.74), swamp bay (r = 0.82), and camphortree (r = 0.87), even severely affected trees of all species were scarcely colonized by the pathogen.

Disease Management in the Genomics Era-Summaries of Focus Issue Papers.

The genomics revolution has contributed enormously to research and disease management applications in plant pathology. This development has rapidly increased our understanding of the molecular mechanisms underpinning pathogenesis and resistance, contributed novel markers for rapid pathogen detection and diagnosis, and offered further insights into the genetics of pathogen populations on a larger scale. The availability of whole genome resources coupled with next-generation sequencing (NGS) technologies has helped fuel genomics-based approaches to improve disease resistance in crops. NGS technologies have accelerated the pace at which whole plant and pathogen genomes have become available, and made possible the metagenomic analysis of plant-associated microbial communities. Furthermore, NGS technologies can now be applied routinely and cost effectively to rapidly generate plant and/or pathogen genome or transcriptome marker sequences associated with virulence phenotypes in the pathogen or resistance phenotypes in the plant, potentially leading to improvements in plant disease management. In some systems, investments in plant and pathogen genomics have led to immediate, tangible benefits. This focus issue covers some of the systems. The articles in this focus issue range from overall perspective articles to research articles describing specific genomics applications for detection and control of diseases caused by nematode, viral, bacterial, fungal, and oomycete pathogens. The following are representative short summaries of the articles that appear in this Focus Issue .

Leaf proteome alterations in the context of physiological and morphological responses to drought and heat stress in barley (Hordeum vulgare L.).

The objective of this study was to identify barley leaf proteins differentially regulated in response to drought and heat and the combined stresses in context of the morphological and physiological changes that also occur. The Syrian landrace Arta and the Australian cultivar Keel were subjected to drought, high temperature, or a combination of both treatments starting at heading. Changes in the leaf proteome were identified using differential gel electrophoresis and mass spectrometry. The drought treatment caused strong reductions of biomass and yield, while photosynthetic performance and the proteome were not significantly changed. In contrast, the heat treatment and the combination of heat and drought reduced photosynthetic performance and caused changes of the leaf proteome. The proteomic analysis identified 99 protein spots differentially regulated in response to heat treatment, 14 of which were regulated in a genotype-specific manner. Differentially regulated proteins predominantly had functions in photosynthesis, but also in detoxification, energy metabolism, and protein biosynthesis. The analysis indicated that de novo protein biosynthesis, protein quality control mediated by chaperones and proteases, and the use of alternative energy resources, i.e. glycolysis, play important roles in adaptation to heat stress. In addition, genetic variation identified in the proteome, in plant growth and photosynthetic performance in response to drought and heat represent stress adaption mechanisms to be exploited in future crop breeding efforts.

Peroxysomal carnitine acetyl transferase influences host colonization capacity in Sclerotinia sclerotiorum.

In lower eukaryotes, the glyoxylate cycle allows cells to utilize two-carbon compounds when simple sugars are not available. In filamentous fungi, glyoxylate metabolism is coupled with ß-oxidation of fatty acids, and both are localized to ubiquitous eukaryotic organelles called peroxisomes. Acetyl coenzyme A (acetyl-CoA) produced during ß-oxidation is transported via the cytosol into mitochondria for further metabolism. A peroxisomal-specific pathway for acetyl-CoA transport requiring peroxisomal carnitine acetyl transferase (CAT) activity has been identified in Magnaporthe grisea peroxisomes. Here, we report that a Sclerotinia sclerotiorum ortholog of the M. grisea peroxisomal CAT-encoding gene Pth2 (herein designated Ss-pth2) is required for virulence-associated host colonization. Null (ss-pth2) mutants, obtained by in vitro transposon mutagenesis, failed to utilize fatty acids, acetate, or glycerol as sole carbon sources for growth. Gene expression analysis of these mutants showed altered levels of transcript accumulation for glyoxylate cycle enzymes. Ss-pth2 disruption also affected sclerotial, apothecial, and appressorial development and morphology, as well as oxalic acid accumulation when cultured with acetate or oleic acid as sole carbon nutrient sources. Although mutants were able to penetrate and initially colonize host tissue, subsequent colonization was impaired. Genetic complementation with the wild-type Ss-pth2 restored wild-type virulence phenotypes. These findings suggest an essential role in S. sclerotiorum for the peroxisomal metabolic pathways for oxalic acid synthesis and host colonization.

Multi-factor regulation of pectate lyase secretion by Colletotrichum gloeosporioides pathogenic on avocado fruits.

Tissue alkalinization during Colletotrichum gloeosporioides attack enhances the expression of PELB, which encodes pectate lyase (PL), and PL secretion, which is considered essential for full virulence. We studied the regulation of PL secretion by manipulation of C. gloeosporioides PELB. PELB was down-regulated by knocking out PAC1, which encodes the PacC transcription factor that regulates gene products with pH-sensitive activities. We functionally characterized a PACC gene homologue, PAC1, from C. gloeosporioides wild-type (WT) Cg-14 and two independent deletion strains, Deltapac1(372)and Deltapac1(761). Loss-of-function PAC1 mutants showed 85% reduction of PELB transcript expression, delayed PL secretion and dramatically reduced virulence, as detected in infection assays with avocado fruits. In contrast, PELB was up-regulated in the presence of carbon sources such as glucose. When glucose was used as a carbon source in the medium for the WT strain and the Deltapac1 mutant at pH 6.0, PELB transcript expression and PL secretion were activated. Other sugars, such as sucrose and fructose (but not galactose), also activated PELB expression. These results suggest that the pH-regulated response is only part of a multi-factor regulation of PELB, and that sugars are also needed to promote the transition from quiescent to active necrotrophic development by the pathogen.

Effect of ammonia production by Colletotrichum gloeosporioides on pelB activation, pectate lyase secretion, and fruit pathogenicity.

The accumulation of ammonia and associated tissue alkalinization predispose avocado fruit to attack by Colletotrichum gloeosporioides. Secretion of ammonia by C. gloeosporioides in the presence of KNO3 was induced by decreasing the pH from 7.0 to 4.0. When the fungus was grown at pH 4.0 or 6.0 in the absence of a nitrogen source, ammonia did not accumulate, and neither pelB (encoding pectate lyase) transcription nor pectate lyase secretion was detected. Under these nitrogen starvation conditions, only transcriptional activation of areA, which encodes the global nitrogen regulator, was detected. pelB transcription and pectate lyase secretion were both detected when C. gloeosporioides was grown at pH 6.0 in the presence of ammonia accumulated from different nitrogen sources. The early accumulation of ammonia induced early pelB expression and pectate lyase secretion. As the external pH increased from 4.0 to 6.0, transcripts of pac1, the C. gloeosporioides pacC homolog, also could be detected. Nit mutants of C. gloeosporioides, which cannot utilize KNO3 as a nitrogen source, did not secrete ammonia, alkalinize the medium, or secrete pectate lyase. If Nit mutants were grown at pH 6.0 in the presence of glutamate, then pectate lyase secretion was induced. Infiltration of 0.1 M ammonium hydroxide at pH 10 into ripening avocado fruits enhanced the activation of quiescent infection and symptom development by C. gloeosporioides. These results suggest that ambient pH alkalinization resulting from ammonia accumulation and the availability of ammonia as a nitrogen source independently regulate pelB expression, pectate lyase secretion, and virulence of C. gloeosporioides. These data suggest that alkalinization during C. gloeosporioides infection is important for its transformation from the quiescent biotrophic stage to the necrotrophic stage of fungal colonization in the fruit host.

External pH and nitrogen source affect secretion of pectate lyase by Colletotrichum gloeosporioides.

Accumulation of ammonia and associated tissue alkalinization predispose fruit to attack by Colletotrichum gloeosporioides: As the external pH increases from 4.0 to 6.0, pectate lyase (PL) and other extracellular proteins are secreted and accumulate. At pH 4.0 neither pelB (encoding PL) transcription nor PL secretion were detected; however, they were detected as the pH increased. Nitrogen assimilation also was required for PL secretion at pH 6.0. Both inorganic and organic nitrogen sources enhanced PL secretion at pH 6.0, but neither was sufficient for PL secretion at pH 4.0. Sequence analysis of the 5' upstream region of the pelB promoter revealed nine putative consensus binding sites for the Aspergillus transcription factor PacC. Consistent with this result, the transcript levels of pac1 (the C. gloeosporioides pacC homologue) and pelB increased in parallel as a function of pH. Our results suggest that the ambient pH and the nitrogen source are independent regulatory factors for processes linked to PL secretion and virulence of C. gloeosporioides.

Longitudinal study of health maintenance organization efficiency.

Longitudinal studies looking at health maintenance organization (HMO) efficiency in the United States have not been performed before this study--previously there have only been small inter-group studies comparing efficiencies. The objectives here were twofold: to analyse HMO efficiency longitudinally, using a data envelopment analysis (DEA) model; and to compare the various types of HMOs and their efficiency using DEA. A cohort of HMOs was followed longitudinally and a number of variables measuring efficiency were studied. Data were derived from the HMO database survey (HCIA Inc.) for 1993-1997. Thirty-six HMOs were followed over a five-year period, and baseline input and output variables were collected. These measures are proxies for efficiency using the DEA methodology. Over all, using the DEA model it was demonstrated that HMOs tended to improve efficiency over time. Independent Practice Association HMOs appeared to show the highest level of improved efficiency. HMO type, profit status, federal eligibility and age were predictive variables for efficiency. Results of this study demonstrate that HMOs are improving efficiency longitudinally, and in general, certain specific models are longitudinally more efficient than others. Moreover, attributes of efficient HMOs have been identified. For policy-makers looking at efficiency, this information can be used to determine which HMO models are more appropriate when trying to achieve cost containment and effectiveness.

Comparison of AIDS progression and survival in persons with pulmonary versus extrapulmonary tuberculosis in Los Angeles.

The objective of this research was to compare the demographics, acquired immune deficiency syndrome (AIDS) progression, and survival in persons with AIDS with pulmonary tuberculosis (PTB) versus extrapulmonary tuberculosis (EPTB), because there are limited population-based data on this topic. A population-based longitudinal study with 3 years of follow-up was performed. Data were collected every 6 months from medical records of persons with AIDS and TB treated at private and public medical facilities throughout Los Angeles County (LAC). Participants included a population-based sample of 216 persons with AIDS and PTB and 166 persons with AIDS and EPTB (including 113 persons with both PTB and EPTB), with an AIDS diagnosis reported in 1993. Compared to persons with AIDS with PTB, persons with AIDS and EPTB were 2.2 times more likely to be Latino than white (95% confidence intervals [CIs]: 1.2, 4.0) and 1.7 times more likely to be foreign-born (95% CIs: 1.1, 2.5). Compared to persons with AIDS with PTB, persons with AIDS and EPTB had similar antiretroviral and PCP prophylaxis use; lower CD4 counts at time of AIDS diagnosis (p = 0.0004); no differences in CD4 counts over the total follow-up period (p = 0.4); higher rates of total opportunistic infections (OIs) (incidence density ratio [IDR] = 2.0; 95% CIs: 1.6, 2.4); and comparable survival curves (p = 0.07). Persons with AIDS and EPTB had a more complicated medical course with lower CD4 counts at time of AIDS diagnosis and more OIs over the follow-up period than persons with AIDS and PTB, however the survival profiles for the two groups were comparable.

T lymphocytes conditioned with Interferon beta induce membrane and soluble VCAM on human brain endothelial cells.

Vascular cell adhesion molecule (VCAM)-1 plays a critical role in mediating inflammatory cell adhesion and migration. Factors regulating the expression of membrane (m)VCAM and its cleaved counterpart soluble (s)VCAM are poorly understood. We previously demonstrated that serum sVCAM levels are increased in multiple sclerosis (MS) patients treated with interferon beta 1b (IFNbeta1b), which correlated with a reduction in gadolinium enhancing lesions on magnetic resonance imaging. However, subsequent studies have shown that IFNbeta does not directly induce VCAM expression on endothelial cells. We demonstrate here that co-culture with IFNbeta-conditioned T cells induces mVCAM on human brain endothelial cells (HBEC). Further, rapid shedding of sVCAM occurs, which mirrors the response after in vivo IFNbeta treatment. The VCAM induction is mediated partially through tumor necrosis factor (TNF)alpha and can be abrogated by sTNF receptor. VCAM could also be induced on astroglioma lines using IFNbeta-conditioned T cells, which suggests the effect is not specific for HBEC. Kinetic studies demonstrated an increase in the sVCAM to mVCAM ratio over time, which may contribute to the ultimate therapeutic effect of IFNbeta in patients. These data have important implications for understanding the events occurring at the blood brain barrier in vivo, and for determining the mechanism of action of IFNbeta in MS.

pH signaling in Sclerotinia sclerotiorum: identification of a pacC/RIM1 homolog.

Sclerotinia sclerotiorum acidifies its ambient environment by producing oxalic acid. This production of oxalic acid during plant infection has been implicated as a primary determinant of pathogenicity in this and other phytopathogenic fungi. We found that ambient pH conditions affect multiple processes in S. sclerotiorum. Exposure to increasing alkaline ambient pH increased the oxalic acid accumulation independent of carbon source, sclerotial development was favored by acidic ambient pH conditions but inhibited by neutral ambient pH, and transcripts encoding the endopolygalacturonase gene pg1 accumulated maximally under acidic culture conditions. We cloned a putative transcription factor-encoding gene, pac1, that may participate in a molecular signaling pathway for regulating gene expression in response to ambient pH. The three zinc finger domains of the predicted Pac1 protein are similar in sequence and organization to the zinc finger domains of the A. nidulans pH-responsive transcription factor PacC. The promoter of pac1 contains eight PacC consensus binding sites, suggesting that this gene, like its homologs, is autoregulated. Consistent with this suggestion, the accumulation of pac1 transcripts paralleled increases in ambient pH. Pac1 was determined to be a functional homolog of PacC by complementation of an A. nidulans pacC-null strain with pac1. Our results suggest that ambient pH is a regulatory cue for processes linked to pathogenicity, development, and virulence and that these processes may be under the molecular regulation of a conserved pH-dependent signaling pathway analogous to that in the nonpathogenic fungus A. nidulans.

Mobile chest radiography: improving image quality.

Mobile examinations present special challenges yet require the same diagnostic quality as exams performed in the radiology department. This article describes a study designed to identify common errors in mobile chest radiography. The authors identify 3 areas of particular concern: rotation, anatomy not centered to the film and distortion. Possible solutions are suggested and discussed.

Has the mammography quality standards act affected the mammography quality in North Carolina?

OBJECTIVE: The United States Food and Drug Administration implemented federal regulations governing mammography under the Mammography Quality Standards Act (MQSA) of 1992. During 1995, its first year in implementation, we examined the impact of the MQSA on the quality of mammography in North Carolina. MATERIALS AND METHODS: All mammography facilities were inspected during 1993-1994, and again in 1995. Both inspections evaluated mean glandular radiation dose, phantom image evaluation, darkroom fog, and developer temperature. Two mammography health specialists employed by the North Carolina Division of Radiation Protection performed all inspections and collected and codified data. RESULTS: The percentage of facilities that met quality standards increased from the first inspection to the second inspection. Phantom scores passing rate was 31.6% versus 78.2%; darkroom fog passing rate was 74.3% versus 88.5%; and temperature difference passing rate was 62.4% versus 86.9%. CONCLUSION: In 1995, the first year that the MQSA was in effect, there was a significant improvement in the quality of mammography in North Carolina. This improvement probably resulted from facilities' compliance with federal regulations.

Anatomic bases for anterior spinal surgery: surgical anatomy of the cervical vertebral body and disc space.

Twenty adult cadaveric cervical spines were sectioned longitudinally through the midline to display longitudinal sections of the vertebral bodies and disc spaces from C3 to T1. Computer-assisted anatomic images were obtained for measurements of the disc spaces and vertebral bodies. Anteroposterior (AP) depth gradually increased from 16.56 +/- 2.21 mm at C3 to 19.32 +/- 2.30 mm at C7. Greater values of AP depth at the inferior endplate were found at C5 (20.75 +/- 2.87 mm) and C6 (20.56 +/- 2.31 mm) compared with the values at C3 (18.26 +/- 1.82 mm), C4 (19.27 +/- 2.88 mm) and C7 (19.21 +/- 3.22 mm). The AP depth at the superior endplate was greater than that at the inferior endplate. The height of the disc space was found to be lowest at the posterior disc space from C2-3 to C7-T1 (2.95 +/- 0.86 mm at C2-3, 2.78 +/- 0.93 mm at C3-4, 2.45 +/- 0.79 mm at C4-5, 2.92 +/- 0.64 mm at C5-6, 2.46 +/- 0.59 mm at C6-7, 2.93 +/- 1.05 mm at C7-T1), when compared to the height of the disc space at the anterior disc space from C2-3 to C7-T1 (4.07 +/- 0.85 mm at C2-3, 4.34 +/- 1.18 mm at C3-4, 3.95 +/- 1.37 mm at C4-5, 3.55 +/- 1.37 mm at C5-6, 3.55 +/- 0.76 mm at C6-7, 3.67 +/- 1.17 mm at C7-T1). The mid-axis of the disc space was situated at approximately 3 mm above the anterior midpoint of the annulus fibrosus at the level of the lower cervical spine. To reach the posterior portion of the disc space from the anterior midpoint of the annulus fibrosus, a 5 degrees cephalad angulation of the drill relative to the mid-axis of the disc space is necessary. All these original data from cadavers may be helpful during anterior approach for discectomy, vertebrectomy and anterior screw-plate placement.

Cyclooxygenase-1 in human Alzheimer and control brain: quantitative analysis of expression by microglia and CA3 hippocampal neurons.

Epidemiological and clinical studies suggest that nonsteroidal anti-inflammatory drugs (NSAIDs) that inhibit cyclooxygenase (COX) slow the progression and delay the onset of Alzheimer disease (AD). Two isoforms of cyclooxygenase have been identified. Although much effort has recently been focused on the inducible COX-2 isoform, little is known about COX-1 expression in human brain. We report that COX-1 message and immunoreactivity are localized to human hippocampal CA3 and CA4 neurons, granular neurons in neocortical layer IV, and occasional cortical pyramidal neurons. Quantitative in situ hybridization showed no differences between COX-1 mRNA levels in control and AD CA3 hippocampal neurons. COX-1 immunoreactivity was also present in microglial cells in gray and white matter in all brain regions examined. COX-1 appeared to be expressed in microglial cells regardless of their activation state as determined by HLA-DR immunostaining. However, COX-1 immunopositive microglia were found in association with Abeta plaques, and the density of COX-1 immunopositive microglia in AD fusiform cortex was increased. This pattern suggests an overall increase of COX-1 expression in AD. Currently used NSAIDs inhibit both isoforms of cyclooxygenase. The present study shows that COX-1 is widely expressed in human brain, and raises the possibility that COX-1 may contribute to CNS pathology.