A retrospective evaluation of supine pelvic radiography image quality using centring points and anatomical axial rotation, including reliability of measurements (ARLEX-P STUDY).

INTRODUCTION: Pelvic radiographs are commonly used for the investigation of a variety of conditions. Comparison between examinations requires a consistent radiographic technique but variations in image quality and radiographic centring points are frequently reported in the literature. The aim of this study was to establish the amount of variation in the radiographic centring point (RCP) and pelvic axial rotation (PAR), with a secondary aim of reporting the reliability of such measures. METHODS: Using a previously acquired imaging archive, 633 adult pelvis/hip radiographs were identified on a Picture Archiving and Communication System (PACS). Radiographs with bilateral prostheses, evidence of acute pelvic trauma, projections acquired on a stretcher/trolley and those demonstrating large discontinuity between the detector and X-ray field centre were excluded. To determine centring point variation (+ values denote superior variations) and axial rotation multiple measurements were obtained from each radiograph. A video was used to train five observers and each of these reviewed ten random cases to determine inter- and intra-rater reliability. One of the five observers then performed the measurements on all remaining radiographs. RESULTS: Following exclusions 380 radiographs were evaluated. The median (IQR) RCP deviation from the inter-acetabular line was +22 (+2 to +43) mm where both iliac crests were present and -29 (-45 to -12) mm where they were not. Eleven (3%) cases demonstrate RCP variation from the midline of greater than 25 mm (no bias towards the left or right side). The median (IQR) PAR was 0.0 (-1.5 to 1.4) degrees with greater variance in PAR for male participants (p = 0.004). Almost 60% of inter-rater ICC measurements were categorised as excellent, good or moderate. CONCLUSION: Variations in RCP and PAR exist when evaluating a sample of routinely acquired pelvis radiographs. Some initial factors, such as sex and sub-examination type (full pelvis [XPEL] or low centred pelvis [XHIPB]) have been identified as having a statistical affect on variability. Further research and methods to standardise radiographic techniques is required and must be multidimensional in nature. IMPLICATIONS FOR PRACTICE: Selection of radiographic technique, including RCP, appears to influence components of the pelvis radiograph. Given the increasing clinical requirements for pelvic radiography further standardisation alongside individual optimisation is warranted.

Chest CT in patients with shortness of breath: Comparing high pitch CT and conventional CT on respiratory artefacts and dose.

INTRODUCTION: To investigate chest respiratory artefact reduction using High Pitch Dual Source Computed Tomography (HPCT) compared to conventional CT (CCT) in symptomatic patients with shortness of breath. METHODS: Forty patients were prospectively examined on a second-generation Dual Source scanner. They were randomly divided into two groups: twenty patients underwent an experimental HPCT protocol and twenty control cases CCT protocol. Respiratory artefacts were evaluated using an ordinal score (0, 1 and 2) assigned by two readers with five and thirty years of experience. A qualitative assessment was performed using two categorical groups, group 1 = acceptable and group 2 = unacceptable. Dose Length Product (DLP) was compared. RESULTS: The two groups showed a statistical difference in artefacts reduction (p < 0.0001). HPCT demonstrated no artefacts in 82% of cases, while CCT showed no artefacts in 39% of cases. DLP showed no statistical differences (p = 0.6) with mean = 266.9 for HPCT and mean = 282.65 for CCT. HPCT provides high table speed in the z-direction allowing a high temporal resolution, which reduces respiratory artefacts during free-breathing acquisition. Despite the use of two x-ray tubes, the HPCT did not increase the dose to the patient but provided the highest images quality. CONCLUSIONS: In the emergency setting, HPCTs have been critical for achieving good image quality in uncooperative patients. IMPLICATIONS FOR PRACTICE: Acute respiratory failure is a common emergency department presentation, and the choice of high-speed acquisition CT may increase image quality.

Modifications to mobile chest radiography technique during the COVID-19 pandemic - implications of X-raying through side room windows.

INTRODUCTION: Modifications to common radiographic techniques have resulted from the challenges presented by the COVID-19 pandemic. Reports exist regarding the potential benefits of undertaking mobile radiography through side room windows. The aim of this study was to evaluate the impact on image quality and exposure factors when undertaking such examinations. METHODS: A phantom based study was undertaken using a digital X-ray room. Control acquisitions, using a commercially available image quality test tool, were performed using standard mobile chest radiography acquisition factors. Image quality (physical and visual), incidence surface air kerma (ISAK), Exposure Index (EI) and Deviation Index (DI) were recorded. Image quality and radiation dose were further assessed for two additional (experimental) scenarios, where a side room window was located immediately adjacent to the exit port of the light beam diaphragm. The goal of experimental scenario one was to modify exposure factors to maintain the control ISAK. The goal of experimental scenario two was to modify exposure factors to maintain the control EI and DI. Dose and image quality data were compared between the three scenarios. RESULTS: To maintain the pre-window (control) ISAK (76 µGy), tube output needed a three-fold increase (90 kV/4 mAs versus 90 kV/11.25 mAs). To maintain EI/DI a more modest increase in tube output was required (90 kV/8 mAs/ISAK 54 µGy). Physical and visual assessments of spatial resolution and signal-to-noise ratio were indifferent between the three scenarios. There was a slight statistically significant reduction in contrast-to-noise ratio when imaging through the glass window (2.3 versus 1.4 and 1.2; P = 0.005). CONCLUSION: Undertaking mobile X-ray examinations through side room windows is potentially feasible but does require an increase in tube output and is likely to be limited by minor reductions in image quality. IMPLICATIONS FOR PRACTICE: Mobile examinations performed through side room windows should only be used in limited circumstances and future clinical evaluation of this technique is warranted.

First Report of Septoria sp. Infecting Stevia rebaudiana in France and Screening of Stevia rebaudiana Genotypes for Host Resistance.

Stevia rebaudiana, for which cultivation is on the increase worldwide, accumulates acaloric intense sweeteners called steviol glycosides (SGs) in its leaves. Yields can be affected by Septoria leaf spot (SLS) caused by Septoria spp. The objectives of the research were (1) to morphologically and genetically characterize five isolates of Septoria sp. found for the first time from outbreaks of Septoria in stevia fields in Southwestern France and (2) to screen S. rebaudiana germplasm from diverse origins through an automated inoculation method using one of the isolates. Multilocus sequence typing grouped the five isolates obtained from symptomatic plants, closely related to Septoria lycopersici and Septoria apiicola. The response to Septoria sp. of 10 genotypes from different origins was assessed for disease severity (DS), either by visually scoring the symptomatic portion of the whole plants or the portion of symptomatic foliar area (PLSA) determined by image analysis, and the area under the disease progress curve (AUDPC) calculated on the basis of the disease severity rating taken 12, 15, 18, and 21 days after inoculation. No genotypes with complete resistance were identified. Moderately susceptible genotypes "Gawi" and "Esplac1" exhibited only 10 to 15% of symptomatic part on whole plant and the slowest disease development. They could be distinguished from highly susceptible ones "E8", "C", and "E161718" exhibiting up to 40% of symptomatic part on whole plant. The variability of response to Septoria sp. that exists in S. rebaudiana opens up the field of breeding strategies for the development of new cultivars for sustainable and organic S. rebaudiana production.

Anatomical and/or pathological predictors for the "incorrect" classification of red dot markers on wrist radiographs taken following trauma.

OBJECTIVE: To establish the prevalence of red dot markers in a sample of wrist radiographs and to identify any anatomical and/or pathological characteristics that predict "incorrect" red dot classification. METHODS: Accident and emergency (A&E) wrist cases from a digital imaging and communications in medicine/digital teaching library were examined for red dot prevalence and for the presence of several anatomical and pathological features. Binary logistic regression analyses were run to establish if any of these features were predictors of incorrect red dot classification. RESULTS: 398 cases were analysed. Red dot was "incorrectly" classified in 8.5% of cases; 6.3% were "false negatives" ("FNs")and 2.3% false positives (FPs) (one decimal place). Old fractures [odds ratio (OR), 5.070 (1.256-20.471)] and reported degenerative change [OR, 9.870 (2.300-42.359)] were found to predict FPs. Frykman V [OR, 9.500 (1.954-46.179)], Frykman VI [OR, 6.333 (1.205-33.283)] and non-Frykman positive abnormalities [OR, 4.597 (1.264-16.711)] predict "FNs". Old fractures and Frykman VI were predictive of error at 90% confidence interval (CI); the rest at 95% CI. CONCLUSION: The five predictors of incorrect red dot classification may inform the image interpretation training of radiographers and other professionals to reduce diagnostic error. Verification with larger samples would reinforce these findings. ADVANCES IN KNOWLEDGE: All healthcare providers strive to eradicate diagnostic error. By examining specific anatomical and pathological predictors on radiographs for such error, as well as extrinsic factors that may affect reporting accuracy, image interpretation training can focus on these "problem" areas and influence which radiographic abnormality detection schemes are appropriate to implement in A&E departments.

Evolution of Pseudomonas aeruginosa virulence in infected patients revealed in a Dictyostelium discoideum host model.

Pseudomonas aeruginosa can cause acute lung infections in intubated patients or chronic infections in patients with cystic fibrosis (CF). In both situations, P. aeruginosa accumulates specific mutations, in particular in the lasR quorum-sensing regulator gene. Using a Dictyostelium discoideum amoeba model, we assessed whether these mutations affect bacterial virulence. Among a collection of clinical isolates from 16 CF patients, initial isolates were fully virulent in 15 patients, but for late isolates collected several years later, virulence was decreased in eight patients. No significant correlation between genetic inactivation of lasR and decreased virulence was observed. Among strains isolated from ten colonized intubated patients, all initial isolates were fully virulent. Despite the accumulation of lasR-inactivating mutations in strains collected over a 3-week period, no decrease in virulence was observed in eight of 10 patients. In one intubated patient, the virulent initial strain was replaced a few days later with a different, less virulent, strain. We observed a gradual decrease in bacterial virulence in only one intubated patient. We conclude that adaptation of P. aeruginosa to chronically infected CF patients can lead to a slow and gradual loss of virulence, as measured in a Dictyostelium model system. However, loss of virulence is not caused predominantly by mutations in lasR. During short-term colonization of intubated patients for up to 20 days, a decrease in virulence was exceptional, despite the accumulation of lasR mutations.

The determinant of potyvirus ability to overcome the RTM resistance of Arabidopsis thaliana maps to the N-terminal region of the coat protein.

In Arabidopsis thaliana Columbia (Col-0) plants, the restriction of Tobacco etch virus (TEV) long-distance movement involves at least three dominant RTM (restricted TEV movement) genes named RTM1, RTM2, and RTM3. Previous work has established that, while the RTM-mediated resistance is also effective against other potyviruses, such as Plum pox virus (PPV) and Lettuce mosaic virus (LMV), some isolates of these viruses are able to overcome the RTM mechanism. In order to identify the viral determinant of this RTM-resistance breaking, the biological properties of recombinants between PPV-R, which systemically infects Col-0, and PPV-PSes, restricted by the RTM resistance, were evaluated. Recombinants that contain the PPV-R coat protein (CP) sequence in an RTM-restricted background are able to systemically infect Col-0. The use of recombinants carrying chimeric CP genes indicated that one or more PPV resistance-breaking determinants map to the 5' half of the CP gene. In the case of LMV, sequencing of independent RTM-breaking variants recovered after serial passages of the LMV AF199 isolate on Col-0 plants revealed, in each case, amino acid changes in the CP N-terminal region, close to the DAG motif. Taken together, these findings demonstrate that the potyvirus CP N-terminal region determines the outcome of the interaction with the RTM-mediated resistance.

High-resolution mapping and chromosome landing at the root-know nematode resistance locus Ma from Myrobalan plum using a large-insert BAC DNA library.

The Ma gene for root-knot nematode (RKN)resistance from Myrobalan plum (Prunus cerasifera L.)confers a complete-spectrum and a heat-stable resistance to Meloidogvne spp., conversely to Mi-I from tomato,which has a more restricted spectrum and a reduced efficiency at high temperature. This gene was identified from a perennial self-incompatible near-wild rootstock species and lies in cosegregation with the SCAR marker SCAFLP2 on the Prunus linkage group 7 in a 2.3 cM interval between the SCAR SCAL19 and SSR pchgms6 markers. We initiated a map-based cloning of Ma and report here the strategy that rapidly led to fine mapping and direct chromosome landing at the locus. Three pairs of bulks, totaling 90 individuals from half-sibling progenies derived from the Ma-heterozygous resistant accession P.2175, were constructed using mapping data, and saturation of the Ma region was performed by bulked segregant analysis (BSA) of 320 AFLP primer pair combinations. The closest three AFLP markers were transformed into codominant SCARs or CAPS designatedSCAFLP3, SCAFLP4 and SCAFLP5. By completing the mapping population up to 1,332 offspring from P.2175,Ma and SCAFLP2 were mapped in a 0.8 cM interval between SCAFLP3 and SCAFLP4. A large-insert bacterial artificial chromosome (BAC) DNA library of P.2175,totaling 30,720 clones with a mean insert size of 145 kb and a 14-15x Prunus haploid genome coverage was constructed and used to land on the Ma spanning interval with few BAC clones. As P.2175 is heterozygous for the gene, we constructed the resistant and susceptible physical contigs by PCR screening of the library with codominant markers. Additional microsatellite markers were then designed from BAC subcloning or BAC end sequencing. In the resistant contig, a single 280 kb BAC clone was shown to carry the Ma gene; this BAC contains two flanking markers on each side of the gene as well as two cosegregating markers. These results should allow future cloning of the Ma gene in this perennial species.

Microsatellite genetic linkage maps of myrobalan plum and an almond-peach hybrid--location of root-knot nematode resistance genes.

Inheritance and linkage studies were carried out with microsatellite [or simple sequence repeat (SSR)] markers in a F(1) progeny including 101 individuals of a cross between Myrobalan plum ( Prunus cerasifera Ehrh) clone P.2175 and the almond (Prunus dulcis Mill.)-peach ( Prunus persica L. Batsch) hybrid clone GN22 ["Garfi" (G) almond x "Nemared" (N) peach]. This three-way interspecific Prunus progeny was produced in order to associate high root-knot nematode (RKN) resistances from Myrobalan and peach with other favorable traits for Prunus rootstocks from plum, peach and almond. The RKN resistance genes, Ma from the Myrobalan plum clone P.2175 and R(MiaNem) from the 'N' peach, are each heterozygous in the parents P.2175 and GN22, respectively. Two hundred and seventy seven Prunus SSRs were tested for their polymorphism. One genetic map was constructed for each parent according to the "double pseudo-testcross" analysis model. The Ma gene and 93 markers [two sequence characterized amplified regions (SCARs), 91 SSRs] were placed on the P.2175 Myrobalan map covering 524.8 cM. The R(MiaNem) gene, the Gr gene controlling the color of peach leaves, and 166 markers (one SCAR, 165 SSRs) were mapped to seven linkage groups instead of the expected eight in Prunus. Markers belonging to groups 6 and 8 in previous maps formed a single group in the GN22 map. A reciprocal translocation, already reported in a G x N F(2), was detected near the Gr gene. By separating markers from linkage groups 6 and 8 from the GN22 map, it was possible to compare the eight homologous linkage groups between the two maps using the 68 SSR markers heterozygous in both parents (anchor loci). All but one of these 68 anchor markers are in the same order in the Myrobalan plum map and in the almond-peach map, as expected from the high level of synteny within Prunus. The Ma and R(MiaNem)genes confirmed their previous location in the Myrobalan linkage group 7 and in the GN22 linkage group 2, respectively. Using a GN22 F(2) progeny of 78 individuals, a microsatellite map of linkage group 2 was also constructed and provided additional evidence for the telomeric position of R(MiaNem) in group 2 of the Prunus genome.

A set of simple-sequence repeat (SSR) markers covering the Prunus genome.

A set of 109 microsatellite primer pairs recently developed for peach and cherry have been studied in the almond x peach F(2) progeny previously used to construct a saturated Prunus map containing mainly restriction fragment length polymorphism markers. All but one gave amplification products, and 87 (80%) segregated in the progeny and detected 96 loci. The resulting Prunus map contains a total of 342 markers covering a total distance of 522 cM. The approximate position of nine additional simple sequence repeats (SSRs) was established by comparison with other almond and peach maps. SSRs were placed in all the eight linkage groups of this map, and their distribution was relatively even, providing a genome-wide coverage with an average density of 5.4 cM/SSR. Twenty-four single-locus SSRs, highly polymorphic in peach, and each falling within 24 evenly spaced approximately 25-cM regions covering the whole Prunus genome, are proposed as a 'genotyping set' useful as a reference for fingerprinting, pedigree and genetic analysis of this species.

Development of microsatellite markers in peach [ Prunus persica (L.) Batsch] and their use in genetic diversity analysis in peach and sweet cherry ( Prunus avium L.).

We report the sequence of 41 primer pairs of microsatellites from a CT-enriched genomic library of the peach cultivar 'Merrill O'Henry'. Ten microsatellite-containing clones had sequences similar to plant coding sequences in databases and could be used as markers for known functions. For microsatellites segregating at least in one of the two Prunus F(2) progenies analyzed, it was possible to demonstrate Mendelian inheritance. Microsatellite polymorphism was evaluated in 27 peach and 21 sweet cherry cultivars. All primer pairs gave PCR-amplification products on peach and 33 on cherry (80.5%). Six PCR-amplifications revealed several loci (14.6%) in peach and eight (19.5%) in sweet cherry. Among the 33 single-locus microsatellites amplified in peach and sweet cherry, 13 revealed polymorphism both in peach and cherry, 19 were polymorphic only on peach and one was polymorphic only on cherry. The number of alleles per locus ranged from 1 to 9 for peach and from 1 to 6 on sweet cherry with an average of 4.2 and 2.8 in peach and sweet cherry, respectively. Cross-species amplification was tested within the Prunus species: Prunus avium L. (sweet cherry and mazzard), Prunus cerasus L. (sour cherry), Prunus domestica L. (European plum), Prunus amygdalus Batsch. (almond), Prunus armeniaca L. (apricot), Prunus cerasifera Ehrh. (Myrobalan plum). Plants from other genera of the Rosaceae were also tested: Malus (apple) and Fragaria (strawberry), as well as species not belonging to the Rosaceae: Castanea (chestnut tree), Juglans (walnut tree) and Vitis (grapevine). Six microsatellites gave amplification on all the tested species. Among them, one had an amplified region homologous to sequences encoding a MADS-box protein in Malus x domestica. Twelve microsatellites (29.3%) were amplified in all the Rosaceae species tested and 31 (75.6%) were amplified in all the six Prunus species tested. Thirty three (80.5%), 18 (43.9%) and 13 (31.7%) gave amplification on chestnut tree, grapevine and walnut tree, respectively.

Candidate genes and QTLs for sugar and organic acid content in peach [ Prunus persica (L.) Batsch].

The identification of genes involved in variation of peach fruit quality would assist breeders in creating new cultivars with improved fruit quality. Major genes and quantitative trait loci (QTLs) for physical and chemical components of fruit quality have already been detected, based on the peach [ Prunus persica (L.) Batsch] cv. Ferjalou Jalousia((R)) (low-acid peach) x cv. Fantasia (normally-acid nectarine) F(2) intraspecific cross. Our aim was to associate these QTLs to structural genes using a candidate gene/QTL approach. Eighteen cDNAs encoding key proteins in soluble sugar and organic acid metabolic pathways as well as in cell expansion were isolated from peach fruit. A single-strand conformation polymorphism strategy based on specific cDNA-based primers was used to map the corresponding genes. Since no polymorphism could be detected in the Ferjalou Jalousia((R)) x Fantasia population, gene mapping was performed on the almond [ Prunus amygdalus ( P. dulcis)] cv. Texas x peach cv. Earlygold F(2) interspecific cross from which a saturated map was available. Twelve candidate genes were assigned to four linkage groups of the peach genome. In a second step, the previous QTL detection was enhanced by integrating anchor loci between the Ferjalou Jalousia((R)) x Fantasia and Texas x Earlygold maps and data from a third year of trait assessment on the Ferjalou Jalousia((R)) x Fantasia population. Comparative mapping allowed us to detect a candidate gene/QTL co-location. It involved a cDNA encoding a vacuolar H(+)-pyrophosphatase ( PRUpe;Vp2) that energises solute accumulation, and QTLs for sucrose and soluble solid content. This preliminary result may be the first step in the future development of marker-assisted selection for peach fruit sucrose and soluble solid content.

Localization of the Rh50-like protein to the contractile vacuole in Dictyostelium.

The human Rhesus (Rh) family consists of three polytopic membrane proteins present at the cell surface of red blood cells. Although Rh proteins are essential for the expression of the blood group system their biological function remains unclear. In this study, the gene encoding a protein homologous to Rh50 in Dictyostelium discoideum was sequenced. The Rh50-like protein was localized to the contractile vacuole, the organelle responsible for maintenance of osmotic equilibrium within the cell. However, Rh50-like-deficient mutants in which the Rh50-like gene was disrupted did not appear to exhibit a phenotype related to osmoregulation. Nevertheless, these mutants may provide a valuable tool for studying the function of the rhesus protein.