Development and application of a nonradioactive nucleic acid hybridization system for simultaneous detection of four potato pathogens

cDNA clones of potato virus X (PVXcp strain), potato virus Y (PVYo strain), potato leaf roll virus (PLRV) and potato spindle tuber viroid (PSTV) were used separately or combined for the detection of the corresponding RNAs in extracts of infected plants. A general method for the rapid preparation of RNA extracts without use of organic solvents (i.e. phenol) was developed for this purpose. Plant extracts from a range of field, artificially inoculated germplasm genotypes, micro-propagated and protoplast samples, as well as vector insect extracts, were dot-blotted onto nylon or nitrocellulose membranes, subjected to sandwich nucleic acid hybridization with non-labelled specific single-stranded DNA probes followed by a biotin-labelled second step hybridization probe. Each probe was virus-specific but not strain-specific. Healthy or non-related plant extracts developed very faint or no signals. Sensitivity was tested by slot-blot hybridization. Detection levels were between 1.5 to 6 pg of viral nucleic acids and between 20 to 50 times more sensitive than standard double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA). The assay developed was tested with material that was prepared for processing in the field (combination of fresh sap with extraction solution) and tested under simple laboratory conditions for detection. It was also successfully employed for screening of germplasm for virus resistance, detection of pathogens in vector insects, plantlets grown in vitro and in more sophisticated quantitative determinations of viral replication in artificially inoculated plants and protoplasts.     

Reframing of an addiction via hypnotherapy: a case presentation

A chemically dependent man was treated using hypnotherapy and related psychotherapeutic techniques. I started by taking a psychosocial history and then introduced such interventions as pattern interruption and symbolic task assignments to establish initial sobriety. The majority of the sessions focused on age regressing the patient to events correlating to drug and alcohol abuse. During these events I introduced myself via hypnosis as "the voice from the future" to redefine the events and extract the useful learnings. With a new-found positive self-image, the patient was hypnotically age progressed to review future scenes. In each scene he successfully abstained from drug and alcohol use. The patient remained drug and alcohol free during treatment and the 6-month and one-year follow-ups.     

The association between the etiology of a spinal cord injury and time to mortality in the United States: A 44-year investigation

Objective: To determine the association between spinal cord injury (SCI) etiology categories and mortality, and examine the association between etiology sub-categories and mortality.

Design: Prospective cohort study.

Setting: Model Systems and Shriners Hospital SCI units.

Participants: Data were analyzed from 42,627 cases in the SCI Model System Collaborative Survival Study Database from 1973 to 2017. Those with SCI etiologies categorized as vehicular, violent, sports, falls, pedestrian, and medical were included.

Interventions: Not applicable.

Outcome Measure: Time to mortality after SCI.

Results: Relative to the sports related etiology category, those with medical, pedestrian, violence, falls, and vehicular related SCIs had a 2.00 (95% confidence intervals (CIs): 1.79–2.24), 1.57 (CIs: 1.34–1.83), 1.54 (CIs: 1.41–1.68), 1.35 (CIs: 1.25–1.45), and 1.26 (CIs: 1.17–1.35) higher hazard for mortality, respectfully. Persons with SCIs from automobile crashes had a 1.38 (CIs: 1.23–1.56) higher hazard for mortality, whereas those with SCIs from motorcycle crashes had a 1.21 (CIs: 1.04–1.39) higher hazard for mortality, relative to other etiologies within the vehicular category. Those with SCIs from diving had a 1.37 (CIs: 1.18–1.59) higher hazard for mortality relative to other etiologies within the sports category.

Conclusions: Injury etiology categories and certain sub-categories were associated with a higher risk for early mortality. Understanding how additional factors such as socioeconomic status, co-occurring injuries, medical co-morbidities, and environmental aspects interact with SCI etiologies may provide insights for how etiology of injury impacts survival. These findings may serve as a development for extending long-term life expectancy by informing SCI prevention programs and care post-injury.     

Analysis of junior doctor supervision in Australasian emergency departments

Background: Supervision of junior doctors in ED is vital but limited literature exists on how it is provided. Objective: To assess Australasian ED supervision and review regional legislature supervision requirements. Methods: Between December 2008 and June 2009 emails containing a link to a cross‐sectional survey were sent to Directors of Emergency Medicine Training in all Australasian ED accredited for advanced training. Non‐responding ED were subsequently contacted by telephone or email. Regional legislature supervision requirements were obtained from postgraduate medical councils. Results: A total of 103 (98.1%) of 105 ED participated. Senior review in person was mandatory in 43.2% of ED for patients of PGY1 (postgraduate year 1 doctors) and 6.1% of ED for patients of PGY2 (P < 0.001). Of ED without mandatory review, 13% had written guidelines detailing which patients required review. When ED consultants were on‐site, they most commonly provided supervision in 60.2% of ED and shared supervision equally with registrars in 35.7% of ED; when consultants were off‐site registrars most commonly provided supervision in 87.6% of ED. Fewer major regional/rural base hospitals had 24 h PGY3 or above supervision than major referral and urban district hospitals (82.6% vs 100% and 100%, P < 0.01). Regional legislature requirements varied with no universal guidelines. Conclusion: There are significant differences between supervision requirements for PGY1 and PGY2. A minority of ED in Australasia do not have 24 h supervision by PGY3 or higher. Few ED have written guidelines for supervising PGY1 and PGY2. The majority of registrar supervision occurs without consultant oversight. Legislature requirements for supervision in ED are variable between regions.     

Prevalence and significance of MEFV gene mutations in patients with gouty arthritis

Gouty arthritis is a chronic erosive autoinflammatory disease. Pyrin has anti-inflammatory effects in the regulation of inflammasome and is encoded by the MEFV gene. The relationship between different rheumatic diseases and the MEFV gene mutations was demonstrated. The aim of this study was to determine the frequency of MEFV gene mutations in patients with gouty arthritis and identify a possible correlation with disease phenotype. Ninety-three patients with gouty arthritis and 102 healthy controls, compatible with age, gender and ethnicity, were included in the study. MEFV gene mutations were investigated by PCR method. Out of 93 patients with gouty arthritis, 36 (38.7 %) showed MEFV gene mutations carriage, whereas 20.6 % in healthy control group. Distribution of mutations identified in patients with gouty arthritis was as; R202Q in 18 (19.3 %), E148Q in 5 (5.4 %), K695R in 4 (4.3 %), M680I in 2 (2.1 %), V726A in 2 (2.1 %), P369S in 2 (2.1 %), R408Q in 2 (2.1 %), M694 V in 1 (1.1 %), respectively. Three patients were identified with compound heterozygosity. Distribution of MEFV gene mutations carriage in healthy controls was; E148Q in 11 (10.7 %), M694 V in 2 (1.9 %), M694I in 1 (0.9 %), M680I in 2 (1.9 %), V726A in 1 (0.9 %), A744S in 1 (0.9 %), K695R in 2 (1.9 %), and P369S in 1 (0.9 %) patients, respectively. Higher MEFV gene mutations carrier frequency was observed in patients with gouty arthritis, compared with the control group (p = 0.009). Heterozygous R202Q was the most common mutation detected in patients with gouty arthritis, while heterozygous E148Q in healthy control group. Statistically significant difference was not detected between clinical findings of gouty arthritis and the MEFV gene mutations (p > 0.05). We determined higher prevalence of MEFV gene mutations in patients with gouty arthritis compared with the healthy control group. The most frequently detected mutation was heterozygous R202Q, whereas E148Q in healthy controls. High carriage rates of MEFV gene mutations in gouty arthritis suggest that it may play an important role in the pathogenesis of the disease and predisposition to the disease.     

Pentraxin 3 concentrations of the mothers with preterm premature rupture of membranes and their neonates,and early neonatal outcome

Background: Pentraxin 3 (PTX3) is an acute phase reactant which has been used to detect intra-amniotic infections (IAI) in pregnancy, but the prognostic value of PTX3 concentrations on neonates has not been studied. We aimed to investigate the relationship between maternal PTX3–neonatal PTX3 concentrations and early neonatal outcome.

Methods: The mothers diagnosed with preterm prelabor rupture of membranes (PPROM) (n?=?28) and their preterm infants (n?=?28) were included in the study. PTX3 concentrations were studied in plasma in the maternal peripheral blood and umbilical/peripheral vein in the neonates. The relationship between the mPTX3–nPTX3 concentrations and neonatal outcome were investigated using non-parametric tests and binary logistic regression analysis.

Results: The mean mPTX3 concentration was 10.35?±?7.82?μg/L. Ten (35.7%) of all mothers were within the normal range and 18 (64.3%) in high percentile (≥97.5 percentile). There was no relation between mPTX3 concentrations and clinical or histologic chorioamnionitis, latency of PPROM, and early neonatal outcome. Mean nPTX3 concentrations was 9.18?±?7.83?μg/L and high nPTX3 concentrations were detected in five (17.8%) neonates. nPTX3 concentrations were inversely correlated with gestational age and correlated with rate of intraventricular hemorrhage (IVH) and mortality. Neonates with high nPTX3 concentrations also have lowered APGAR scores, increased rate of respiratory distress syndrome, clinical sepsis, IVH, necrotizing enterocolitis and prolonged NICU stay.

Conclusion: High PTX3 concentrations of the newborns are associated with some worsened early neonatal outcome including lower gestational age at delivery, increased rate of IVH and mortality. Maternal PTX3 concentrations are not an adequate marker in defining clinical or histologic chorioamnionitis and early neonatal outcome.