Real-time PCR detection and quantification of vector trichodorid nematodes and Tobacco rattle virus

This report describes a novel diagnostic method for virus-vector trichodorid nematodes and associated Tobacco rattle virus (TRV) based on a real-time fluorogenic 5' nuclease PCR assay (TaqMan). Two independent primer/probe sets were designed targeting the 18S gene of the ribosomal cistron for the trichodorid species, Paratrichodorus pachydermus and Trichodorus similis. Assays using purified plasmid DNA containing clones of the 18S region and genomic DNA extracted from individuals from both nematode species displayed high specificity as no cros s-reaction was observed between the species or with two non-target trichodorid species Paratrichodorus anemones and Trichodorus primitivus. Relative quantification of target DNA present in unknown samples was performed by comparison of the fluorescence signals of the samples to those obtained from plasmid standard dilutions. Three primer/probe sets were also used to target TRV; one set for RNA1 and the two other sets for RNA2 of specific isolates (TRV-PpK20 and TRV-TpO1). Detection of both trichodorid species and TRV RNA1 and RNA2 from a single sample was achieved and field samples were used to demonstrate the potential of this assay to provide rapid, accurate and sensitive molecular information in relation to risk assessment in the field.     

Genome sequence of the enterobacterial phytopathogen Erwinia carotovora subsp. atroseptica and characterization of virulence factors

The bacterial family Enterobacteriaceae is notable for its well studied human pathogens, including Salmonella, Yersinia, Shigella, and Escherichia spp. However, it also contains several plant pathogens. We report the genome sequence of a plant pathogenic enterobacterium, Erwinia carotovora subsp. atroseptica (Eca) strain SCRI1043, the causative agent of soft rot and blackleg potato diseases. Approximately 33% of Eca genes are not shared with sequenced enterobacterial human pathogens, including some predicted to facilitate unexpected metabolic traits, such as nitrogen fixation and opine catabolism. This proportion of genes also contains an overrepresentation of pathogenicity determinants, including possible horizontally acquired gene clusters for putative type IV secretion and polyketide phytotoxin synthesis. To investigate whether these gene clusters play a role in the disease process, an arrayed set of insertional mutants was generated, and mutations were identified. Plant bioassays showed that these mutants were significantly reduced in virulence, demonstrating both the presence of novel pathogenicity determinants in Eca, and the impact of functional genomics in expanding our understanding of phytopathogenicity in the Enterobacteriaceae.     

Psychometric Properties of the Greek Version of FCV-19S

As COVID-19 delivered a devastating blow around Europe, unprecedented mental health issues have arisen. This study outlines the Greek translation and validation of the Fear of COVID-19 Scale (FCV-19S) in the general population. A sample of 3029 Greek-speaking individuals completed the FCV-19S, as part of a large online survey reporting on the aftermath of the coronavirus on the psychological health of Greeks. A total of 2970 participants completed all items without missing values, and their responses were included in the statistical analysis. The confirmatory factor analysis suggested an acceptable model fit. The internal consistency measured by Cronbach’s alpha = .87 for the whole scale. All hypothesized correlations were as expected, confirming the construct validity of the scale. The results of the analyses suggest that the psychometric properties of the Greek FCV-19S are sufficient.

     

Yeast two-hybrid study of tobacco rattle virus coat protein and 2b protein interactions

Summary. The specificity of the interaction between the coat protein (CP) and 2b nematode-transmission helper protein of two isolates, PpK20 and PaY4, of Tobacco rattle virus (TRV) that differ in their transmission characteristics was investigated. Yeast two-hybrid experiments identified a central domain of the CP that promoted CP:CP interaction but inhibited CP:2b interaction. Deletion of a conserved coiled-coil domain from the 2b protein prevented its interaction with CP, whereas deletion of N- and C-terminal domains of the 2b protein greatly enhanced its interaction with CP. A C-terminal flexible domain of the CP was also shown to be important for interaction with the 2b protein. However, this domain was not sufficient to direct isolate-specific interaction of these proteins either in yeast or via a chimeric TRV in planta. Although these two TRV isolates are both transmitted by a shared vector trichodorid nematode, Paratrichodorus pachydermus, the CP from isolate PpK20 did not interact with the 2b protein from isolate PaY4, and vice versa, suggesting that in the field mixed infections of TRV are unlikely to cause promiscuous transmission by alternative trichodorid nematode species.     

Scintigraphic studies of rectal emptying in patients with constipation and defecatory difficulty

We prospectively evaluated 38 adult patients with chronic constipation with and without defecatory difficulties using a newly described scintigraphic test to measure rectal emptying and compared them to 20 healthy controls. All patients underwent anorectal manometry, and 30 who complained of infrequent defecation underwent a colonic transit study using radiopaque markers. Control subjects promptly evacuated both 100 ml and 200 ml artificial stool in a characteristic fashion, but three evacuated none of the 100-ml volume and two had no evacuation of the 200-ml stool (inhibited controls). Constipated patients exhibited three patterns of emptying: (1) normal emptying of both volumes (47%); (2) poor emptying of both volumes or inhibited defecation (29%); and (3) normal emptying of the 200-ml but abnormal evacuation of the 100-ml volume (24%). An abnormal expulsion pattern during manometry occurred in 21% of patients and was strongly associated with the inhibited defecation pattern. However, defecation patterns could not be predicted on the basis of age, gender, symptoms, duration of complaints, colonic transit, or other rectal manometric parameters. Although rectal scintigraphy has potential advantages as a diagnostic test in terms of quantitation and decreased radiation exposure, the inability of the test to distinguish patients with slow transit constipation and defecatory complaints makes the potential utility of this test of uncertain value in clinical and investigative settings.     

Psychological Evaluation of Patients with Severe Idiopathic Constipation: Which Instrument to Use

We prospectively evaluated 38 patients with chronic severe idiopathic constipation who failed to respond to conventional therapy. Colonic transit studies, the Hopkins Symptom Checklist (SCL-90-R), and the Minnesota Multiphasic Personality Inventory (MMPI) were completed. Colonic transit studies identified 23 patients with slow transit and 15 with normal transit. Patients with normal transit constipation scored higher than those with slow transit on every scale of both the SCL-90-R and MMPI, but the SCL-90-R appeared to detect greater differences between the two groups. We conclude that the SCL-90-R is the preferred psychological instrument to evaluate patients with chronic severe constipation who fail to respond to conventional therapy. It provides information similar to the MMPI with which to measure behavioral profiles of patients in an office setting, and it can be completed and scored more rapidly.     

Personality and peritraumatic dissociation in the prediction of PTSD in victims of road traffic accidents.

OBJECTIVES: To investigate the contribution of personality and peritraumatic dissociation in the development of posttraumatic stress disorder (PTSD). METHOD: Victims of road traffic accidents (RTA) were assessed within 2-4 weeks (Time 1) of the accident and again between 4 and 6 months (Time 2). The Eysenck Personality Questionnaire (EPQ) and Peritraumatic Dissociation Experience Questionnaire (PDEQ) were administered at Time 1 and posttraumatic stress symptoms were assessed at Time 2. RESULTS: 265 subjects were assessed at both time points, although neuroticism, psychoticism, and peritraumatic dissociation were significantly correlated with posttraumatic symptoms, only the personality dimensions were independent and significant predictors of subsequent PTSD in a logistic regression. CONCLUSIONS: Consistent with the literature personality measures, especially neuroticism, is associated with the development of PTSD. However, peritraumatic dissociation was not found to be an independent predictor of PTSD.     

The role of microorganisms of the wound in postop complications

Sensitivity of mostly wide-spread pathogenic microorganisms to modern antibiotics of the wide spectrum of action (Tienam, Mefoxin, Cifran), and to antibiotics, widely applied in everyday clinical practice was studied.