Genome Organization of Four Brazilian Xanthomonas albilineans Strains Does Not Correlate with Aggressiveness.

An integrative approach combining genomics, transcriptomics, and cell biology is presented to address leaf scald disease, a major problem for the sugarcane industry. To gain insight into the biology of the causal agent, the complete genome sequences of four Brazilian Xanthomonas albilineans strains with differing virulence capabilities are presented and compared to the GPEPC73 reference strain and FJ1. Based on the aggressiveness index, different strains were compared: Xa04 and Xa11 are highly aggressive, Xa26 is intermediate, and Xa21 is the least, while, based on genome structure, Xa04 shares most of its genomic features with Xa26, and Xa11 share most of its genomic features with Xa21. In addition to presenting more clustered regularly interspaced short palindromic repeats (CRISPR) clusters, four more novel prophage insertions are present than the previously sequenced GPEPC73 and FJ1 strains. Incorporating the aggressiveness index and in vitro cell biology into these genome features indicates that disease establishment is not a result of a single determinant factor, as in most other Xanthomonas species. The Brazilian strains lack the previously described plasmids but present more prophage regions. In pairs, the most virulent and the least virulent share unique prophages. In vitro transcriptomics shed light on the 54 most highly expressed genes among the 4 strains compared to ribosomal proteins (RPs), of these, 3 outer membrane proteins. Finally, comparative albicidin inhibition rings and in vitro growth curves of the four strains also do not correlate with pathogenicity. In conclusion, the results disclose that leaf scald disease is not associated with a single shared characteristic between the most or the least pathogenic strains. IMPORTANCE An integrative approach is presented which combines genomics, transcriptomics, and cell biology to address leaf scald disease. The results presented here disclose that the disease is not associated with a single shared characteristic between the most pathogenic strains or a unique genomic pattern. Sequence data from four Brazilian strains are presented that differ in pathogenicity index: Xa04 and Xa11 are highly virulent, Xa26 is intermediate, and Xa21 is the least pathogenic strain, while, based on genome structure, Xa04 shares with Xa26, and Xa11 shares with X21 most of the genome features. Other than presenting more CRISPR clusters and prophages than the previously sequenced strains, the integration of aggressiveness and cell biology points out that disease establishment is not a result of a single determinant factor as in other xanthomonads.

Vaginal displacement during course of adjuvant radiation for cervical cancer: results from a prospective IG-IMRT study.

OBJECTIVE: To compare internal target volume (ITV) generated using population-based displacements (ITV_study) with empty and full bladder scan fusion (ITV_EBFB) for organ-at-risk (OAR) doses during adjuvant intensity-modulated radiation therapy (IMRT) for cervical cancer. METHODS: From January 2011 to October 2012, patients undergoing IMRT were included. CT simulation was carried out after inserting vault markers. Planning target volume (PTV)_EBFB received 50 Gy per 25 fractions. Pre-treatment megavoltage CT (MVCT) was performed. MVCTs were registered using bony landmarks with Day 1 MVCT. Displacement of the centre of mass of markers was measured along each axis. Directional ITV was calculated using mean ± 2 standard deviations (SDs) (ITV_study). Replanning was performed using PTV study, and OAR doses were compared with PTV_EBFB using Wilcoxon test. RESULTS: A total of 348/386 data sets were evaluable for 16 patients. The median vaginal displacement was 1.2 mm (SD, 1.3 mm), 4.0 mm (SD, 3.5 mm) and 2.8 mm (SD, 3.3 mm) in the mediolateral, superoinferior and anteroposterior directions, respectively. The ITV margins were 4.1, 10.3 and 10.6 mm. ITV_study and ITV_EBFB were 115.2 cm(3) (87.7-152.2 cm(3)) and 151 cm(3) (95.7-277.1 cm(3)) (p < 0.0001), respectively. PTV_study and PTV_EBFB were 814 and 881 cm(3) (p < 0.0001), respectively. Median doses to the bladder were lower with the PTV_study (46.2 Gy vs 43.2 Gy; p = 0.0001), and a similar trend was observed in the volume of the small bowel receiving 40 Gy (68.2 vs 60.1 cm(3); p = 0.09). CONCLUSION: Population-based PTV margins can lead to reduction in OAR doses. ADVANCES IN KNOWLEDGE: Population-based ITV may reduce OAR doses while executing adjuvant IMRT for cervical cancer.

Catechol-O-methyltransferase val158met genotype determines effect of reboxetine on emotional memory in healthy male volunteers.

BACKGROUND: Catechol-O-methyltransferase (COMT) metabolizes catecholamines in the prefrontal cortex (PFC). A common polymorphism in the COMT gene (COMT val158met) has pleiotropic effects on cognitive and emotional processing. The met allele has been associated with enhanced cognitive processing but impaired emotional processing relative to the val allele. METHODS: We genotyped healthy, white men in relation to the COMT val158met polymorphism. They were given a single 4 mg dose of the selective noradrenaline reuptake inhibitor (NRI) reboxetine or placebo in a randomized, double-blind between-subjects model and then completed an emotional memory task 2 hours later. RESULTS: We included 75 men in the study; 41 received reboxetine and 34 received placebo. In the placebo group, met/met carriers did not demonstrate the usual memory advantage for emotional stimuli that was observed in val carriers. Reboxetine restored this emotional enhancement of memory in met/met carriers, but had no significant effect in val carriers. LIMITATIONS: We studied only men, thus limiting the generalizability of our findings. We also relied on self-reported responses to screening questions to establish healthy volunteer status, and in spite of the double-blind design, participants were significantly better than chance at identifying their intervention allocation. CONCLUSION: Emotional memory is impaired in healthy met homozygotes and selectively improved in this group by reboxetine. This has potential translational implications for the use of reboxetine, which is currently licensed as an antidepressant in several countries, and edivoxetine, a new selective NRI currently in development.

Knee muscle strength and visual acuity are the most important modifiable predictors of falls in patients after hip fracture surgery: a prospective study.

Our purpose was to identify risk factors for falls among older adults who had recently undergone hip fracture surgery. The subjects in this study were 69 older adults (aged 65 years or more) who had sustained a hip fracture and were admitted to an orthopedic rehabilitation ward after surgery. Potential fall risk factors were assessed using the physiological profile assessment, timed-up-and-go test, berg balance test, and activities-specific balance confidence scale at discharge from the hospital. Each individual was followed for a period of 6 months to obtain information on the incidence of falls. Receiver operating characteristic curves were constructed to determine the optimal cutoff score for each potential risk factor identified. Multivariate logistic regression was then used to identify the significant predictors of falls and their odds ratios (ORs). During the 6-month follow-up period, 10 of the 69 patients experienced one or more falls. The results showed that fallers were older than nonfallers (p=0.009). Fallers also had poorer performance in the high-contrast visual acuity test (p=0.015) and lower knee flexor (p=0.021) and knee extensor (p=0.005) muscle strength values. Multivariate logistic regression analysis showed that high-contrast visual acuity (cutoff score Z=-2.280, OR=6.14, 95% CI 1.13-33.29, p=0.035) and knee extensor muscle strength (cutoff score Z=-1.835, OR=4.81, 95% CI 1.04-22.33, p=0.045) were predictors of falls. Poor visual acuity and knee muscle weakness are modifiable predictors of falls and should be the key target areas in fall-prevention programs for older adults with hip fractures.

Effect of perfusate Ca2+ on the relation between metabolism and mechanical performance in the rat heart.

Langendorf perfused rat hearts (n = 25) were used to study the effects of changes in perfusate Ca2+ concentration ([Ca2+p]) on the relation between metabolism and mechanical performance with either glucose or pyruvate as the exogenous substrate. Increased [Ca2+p] (from 1.3 to 3.9 mM) produced an increase (243 +/- 38%) in left ventricular developed pressure regardless of the substrate. With glucose as the substrate, the NADH fluorescence intensity increased by 11.8 +/- 1.2% (n = 17) relative to control indicating a more reduced state of the respiratory chain. Increasing [Ca2+p] in the pyruvate perfused heart produced the expected NADH oxidation (-6.2 +/- 1.1%; n = 8). Hence the change in NADH fluorescence associated with increased [Ca2+p] is substrate dependent. The data show that, with glucose as the substrate but not with pyruvate, increases in [Ca2+p] increase the availability of reducing equivalents to the respiratory chain above the level necessary to compensate for the increased demand resulting from the greater contractile performance.

Effect of hemorrhagic shock on the performance, O2-consumption, and ultrastructure of isolated rat hearts.

Rat hearts were isolated from control animals anesthetized and sham treated for 2.5 hours and following 2.5 hours of hemorrhagic hypotension and they were perfused by modified Langendorff technique. Hearts isolated following hemorrhagic hypotension exhibited increased coronary resistance, depressed left ventricular mechanical performance, and significantly increased sensitivity to threefold elevation of perfusate Ca2+ (from 1.3 to 3.9 mM). Electron microscopic examination showed increased permeability of the plasma membrane for lanthanum, zonal lesions of contractile filaments, damage of mitochondria, and dilation of T-tubules and sarcoplasmic reticulum. Experimental results indicate that myocardial damage is of hypoxic/ischemic origin during the procedure used as a model for shock. Mechanical failure of postoligemic hearts may be caused by the combination of zonal lesions of the myofilaments, damage of energy production, and impairment of cellular calcium metabolism.