Detección de patógenos asociados a la enfermedad punta morada en los cultivos de papa y tomate en Guatemala / Detection of pathogens associated to potato purple top disease in potato and tomato crops in Guatemala

La punta morada es una enfermedad que afecta la producción de algunas especies de solanáceas como la papa y el tomate, causando enrollamiento en las puntas de las hojas con una marcada coloración morada, decaimiento temprano de la planta y en la papa se observa tuberización aérea. Como patógenos asociados a la enfermedad se consideran al fitoplasma BLTVA y la bacteria Candidatus Liberibacter solanacearum. Dada la similitud en la sin-tomatología foliar que generan ambos patógenos, es difícil precisar cuál de ellos está implicado en la enfermedad. En Guatemala, existen reportes de la sintomatología típica de punta morada en las principales zonas productoras de papa y tomate, desconociéndose el agente asociado. La investigación determinó cuál de los dos patógenos reportados está asociados a la enfermedad en 12 municipios productores de papa y/o tomate en el país. Se realizaron ampli-ficaciones de ADN con cebadores específicos para cada patógeno asociado a la enfermedad. Por la alta incidencia del fitoplasma BLTVA en las muestras de papa (73.9%), en comparación a C. Liberibacter solanacearum (26%), este es considerado como el patógeno asociado más importante en papa. En las muestras de tomate, la incidencia del fitoplasma BLTVA (29.8%) y C. Liberibacter solanacearum del (27.6%) fue similar. Además, sobresale el primer reporte de la detección del fitoplasma BLTVA afectando el cultivo de tomate en Guatemala. Se sugiere un monitoreo constante, mediante métodos moleculares, para un diagnóstico certero y establecer medidas de manejo de la enfermedad para evitar su diseminación hacia zonas aún no afectadas.

The potato purple top is a disease that affects the production of some solanaceous species such as potatoes and tomatoes, causing curl at the tips of the leaves with a marked purple coloration, early decay of the plant, and aerial tuberization is observed in the potato. BLTVA phytoplasma and Candidatus Liberibacter solanacearum are considered as pathogens associated with the disease. Given the similarity in foliar symptoms generated by both pathogens, it is difficult to determine which one is involved in the disease. There are reports of the typical potato purple top symptoms in the main potato and tomato producing areas in Guatemala, being unknown the associated agent. The research determined which of the two reported pathogens is associated with the disease in 12 potatoes and/or tomato producing areas in the country. We performed DNA amplification with specific primers for each disease-associated pathogen. Due to the high incidence of BLTVA phytoplasma in potato samples (73.9%), com-pared to C. liberibacter solanacearum (26%), this is considered the most important associated pathogen in potatoes. In tomato samples, the incidence of BLTVA phytoplasma (29.8%) and C. liberibacter solanacearum (27.6%) was similar. Besides, the first report of the detection of the BLTVA phytoplasma affecting tomato cultivation in Gua-temala stands out. Using molecular methods, constant monitoring is suggested for an accurate diagnosis and to establish management measures for the disease to prevent its spread to areas not yet affected.

Correction: The effect of a programme to improve men's sedentary time and physical activity: The European Fans in Training (EuroFIT) randomised controlled trial.

[This corrects the article DOI: 10.1371/journal.pmed.1002736.].

The effect of a programme to improve men's sedentary time and physical activity: The European Fans in Training (EuroFIT) randomised controlled trial.

BACKGROUND: Reducing sitting time as well as increasing physical activity in inactive people is beneficial for their health. This paper investigates the effectiveness of the European Fans in Training (EuroFIT) programme to improve physical activity and sedentary time in male football fans, delivered through the professional football setting. METHODS AND FINDINGS: A total of 1,113 men aged 30-65 with self-reported body mass index (BMI) ≥27 kg/m2 took part in a randomised controlled trial in 15 professional football clubs in England, the Netherlands, Norway, and Portugal. Recruitment was between September 19, 2015, and February 2, 2016. Participants consented to study procedures and provided usable activity monitor baseline data. They were randomised, stratified by club, to either the EuroFIT intervention or a 12-month waiting list comparison group. Follow-up measurement was post-programme and 12 months after baseline. EuroFIT is a 12-week, group-based programme delivered by coaches in football club stadia in 12 weekly 90-minute sessions. Weekly sessions aimed to improve physical activity, sedentary time, and diet and maintain changes long term. A pocket-worn device (SitFIT) allowed self-monitoring of sedentary time and daily steps, and a game-based app (MatchFIT) encouraged between-session social support. Primary outcome (objectively measured sedentary time and physical activity) measurements were obtained for 83% and 85% of intervention and comparison participants. Intention-to-treat analyses showed a baseline-adjusted mean difference in sedentary time at 12 months of -1.6 minutes/day (97.5% confidence interval [CI], -14.3-11.0; p = 0.77) and in step counts of 678 steps/day (97.5% CI, 309-1.048; p < 0.001) in favor of the intervention. There were significant improvements in diet, weight, well-being, self-esteem, vitality, and biomarkers of cardiometabolic health in favor of the intervention group, but not in quality of life. There was a 0.95 probability of EuroFIT being cost-effective compared with the comparison group if society is willing to pay £1.50 per extra step/day, a maximum probability of 0.61 if society is willing to pay £1,800 per minute less sedentary time/day, and 0.13 probability if society is willing to pay £30,000 per quality-adjusted life-year (QALY). It was not possible to blind participants to group allocation. Men attracted to the programme already had quite high levels of physical activity at baseline (8,372 steps/day), which may have limited room for improvement. Although participants came from across the socioeconomic spectrum, a majority were well educated and in paid work. There was an increase in recent injuries and in upper and lower joint pain scores post-programme. In addition, although the five-level EuroQoL questionnaire (EQ-5D-5L) is now the preferred measure for cost-effectiveness analyses across Europe, baseline scores were high (0.93), suggesting a ceiling effect for QALYs. CONCLUSION: Participation in EuroFIT led to improvements in physical activity, diet, body weight, and biomarkers of cardiometabolic health, but not in sedentary time at 12 months. Within-trial analysis suggests it is not cost-effective in the short term for QALYs due to a ceiling effect in quality of life. Nevertheless, decision-makers may consider the incremental cost for increase in steps worth the investment. TRIAL REGISTRATION: International Standard Randomised Controlled Trials, ISRCTN-81935608.

The feasibility, acceptability and preliminary testing of a novel, low-tech intervention to improve pre-hospital data recording for pre-alert and handover to the Emergency Department.

BACKGROUND: Poor communication during patient handover is recognised internationally as a root cause of a significant proportion of preventable deaths. Data used in handover is not always easily recorded using ambulance based tablets, particularly in time-critical cases. Paramedics have therefore developed pragmatic workarounds (writing on gloves or scrap paper) to record these data. However, such practices can conflict with policy, data recorded can be variable, easily lost and negatively impact on handover quality. METHODS: This study aimed to measure the feasibility and acceptability of a novel, low tech intervention, designed to support clinical information recording and delivery during pre-alert and handover within the pre-hospital and ED setting. A simple pre and post-test design was used with a historical control. Eligible participants included all ambulance clinicians based at one large city Ambulance Station (n = 69) and all nursing and physician staff (n = 99) based in a city Emergency Department. RESULTS: Twenty five (36%) ambulance clinicians responded to the follow-up survey. Most felt both the pre-alert and handover components of the card were either 'useful-very useful' (n = 23 (92%); and n = 18 (72%) respectively. Nineteen (76%) used the card to record clinical information and almost all (n = 23 (92%) felt it 'useful' to 'very useful' in supporting pre-alert. Similarly, 65% (n = 16) stated they 'often' or 'always' used the card to support handover. For pre-alert information there were improvements in the provision of 8/11 (72.7%) clinical variables. ​ Results from the post-test survey measuring ED staff (n = 37) perceptions of handover demonstrated small (p < 0.05) improvements in handover in 3/5 domains measured. CONCLUSION: This novel low-tech intervention was highly acceptable to ambulance clinician participants, improving their data recording and information exchange processes. However, further well conducted studies are required to test the impact of this intervention on information exchange during pre-alert and handover.

Validation of a Novel Device to Measure and Provide Feedback on Sedentary Behavior.

PURPOSE: Pedometers, which enable self-monitoring of step counts, are effective in facilitating increases in physical activity. Similar devices which provide real-time feedback on sedentary (sitting) behavior are limited. This study aimed to develop and validate a novel device-the SitFIT-which could accurately measure and provide feedback on sedentary behavior and physical activity. METHODS: The SitFIT is a triaxial accelerometer, developed by PAL Technologies, which is worn in the front trouser pocket. This enables tracking of thigh inclination and therefore differentiation between sitting and upright postures, as well as tracking of step count. It has a display to provide user feedback. To determine the validity of the SitFIT for measuring sedentary behavior and step counts, 21 men, age 30 to 65 yr, with body mass index 26.6 ± 3.9 kg·m wore a SitFIT in a front trouser pocket and an activPAL accelerometer attached to their thigh for up to 7 d. Outputs from the SitFIT were compared with the activPAL, which was assumed to provide criterion standard measurements of sitting and step counts. RESULTS: Mean step counts were approximately 4% lower with the SitFIT than activPAL, with correlation between the two methods being very high (r = 0.98) and no obvious bias from the line of equality (regression line, y = 1.0035x + 418.35). Mean sedentary time was approximately 5% higher with the SitFIT than activPAL, correlation between methods was high (r = 0.84), and the equation of the regression line was close to the line of equality (y = 0.8728x + 38.445). CONCLUSIONS: The SitFIT has excellent validity for measurement of free-living step counts and sedentary time and therefore addresses a clear need for a device that can be used as a tool to provide feedback on sedentary behavior to facilitate behavior change.

Feasibility of a real-time self-monitoring device for sitting less and moving more: a randomised controlled trial.

OBJECTIVES: Time spent inactive and sedentary are both associated with poor health. Self-monitoring of walking, using pedometers for real-time feedback, is effective at increasing physical activity. This study evaluated the feasibility of a new pocket-worn sedentary time and physical activity real-time self-monitoring device (SitFIT). METHODS: Forty sedentary men were equally randomised into two intervention groups. For 4 weeks, one group received a SitFIT providing feedback on steps and time spent sedentary (lying/sitting); the other group received a SitFIT providing feedback on steps and time spent upright (standing/stepping). Change in sedentary time, standing time, stepping time and step count was assessed using activPAL monitors at baseline, 4-week follow-up (T1) and 12-week (T2) follow-up. Semistructured interviews were conducted after 4 and 12 weeks. RESULTS: The SitFIT was reported as acceptable and usable and seen as a motivating tool to reduce sedentary time by both groups. On average, participants reduced their sedentary time by 7.8 minutes/day (95% CI -55.4 to 39.7) (T1) and by 8.2 minutes/day (95% CI -60.1 to 44.3) (T2). They increased standing time by 23.2 minutes/day (95% CI 4.0 to 42.5) (T1) and 16.2 minutes/day (95% CI -13.9 to 46.2) (T2). Stepping time was increased by 8.5 minutes/day (95% CI 0.9 to 16.0) (T1) and 9.0 minutes/day (95% CI 0.5 to 17.5) (T2). There were no between-group differences at either follow-up time points. CONCLUSION: The SitFIT was perceived as a useful tool for self-monitoring of sedentary time. It has potential as a real-time self-monitoring device to reduce sedentary and increase upright time.

Study protocol of European Fans in Training (EuroFIT): a four-country randomised controlled trial of a lifestyle program for men delivered in elite football clubs.

BACKGROUND: Lifestyle interventions targeting physical activity, sedentary time and dietary behaviours have the potential to initiate and support behavioural change and result in public health gain. Although men have often been reluctant to engage in such lifestyle programs, many are at high risk of several chronic conditions. We have developed an evidence and theory-based, gender sensitised, health and lifestyle program (European Fans in Training (EuroFIT)), which is designed to attract men through the loyalty they feel to the football club they support. This paper describes the study protocol to evaluate the effectiveness and cost-effectiveness of the EuroFIT program in supporting men to improve their level of physical activity and reduce sedentary behaviour over 12 months. METHODS: The EuroFIT study is a pragmatic, two-arm, randomised controlled trial conducted in 15 football clubs in the Netherlands, Norway, Portugal and the UK (England). One-thousand men, aged 30 to 65 years, with a self-reported Body Mass Index (BMI) ≥27 kg/m(2) will be recruited and individually randomised. The primary outcomes are objectively-assessed changes in total physical activity (steps per day) and total sedentary time (minutes per day) at 12 months after baseline assessment. Secondary outcomes are weight, BMI, waist circumference, resting systolic and diastolic blood pressure, cardio-metabolic blood biomarkers, food intake, self-reported physical activity and sedentary time, wellbeing, self-esteem, vitality and quality of life. Cost-effectiveness will be assessed and a process evaluation conducted. The EuroFIT program will be delivered over 12 weekly, 90-minute sessions that combine classroom discussion with graded physical activity in the setting of the football club. Classroom sessions provide participants with a toolbox of behaviour change techniques to initiate and sustain long-term lifestyle changes. The coaches will receive two days of training to enable them to create a positive social environment that supports men in engaging in sustained behaviour change. DISCUSSION: The EuroFIT trial will provide evidence on the effectiveness and cost-effectiveness of the EuroFIT program delivered by football clubs to their male fans, and will offer insight into factors associated with success in making sustained changes to physical activity, sedentary behaviour, and secondary outcomes, such as diet. ISRCTN: 81935608 . Registered 16 June 2015.

Analysis of wild-species introgressions in tomato inbreds uncovers ancestral origins.

BACKGROUND: Decades of intensive tomato breeding using wild-species germplasm have resulted in the genomes of domesticated germplasm (Solanum lycopersicum) being intertwined with introgressions from their wild relatives. Comparative analysis of genomes among cultivated tomatoes and wild species that have contributed genetic variation can help identify desirable genes, such as those conferring disease resistance. The ability to identify introgression position, borders, and contents can reveal ancestral origins and facilitate harnessing of wild variation in crop breeding. RESULTS: Here we present the whole-genome sequences of two tomato inbreds, Gh13 and BTI-87, both carrying the begomovirus resistance locus Ty-3 introgressed from wild tomato species. Introgressions of different sizes on chromosome 6 of Gh13 and BTI-87, both corresponding to the Ty-3 region, were identified as from a source close to the wild species S. chilense. Other introgressions were identified throughout the genomes of the inbreds and showed major differences in the breeding pedigrees of the two lines. Interestingly, additional large introgressions from the close tomato relative S. pimpinellifolium were identified in both lines. Some of the polymorphic regions were attributed to introgressions in the reference Heinz 1706 genome, indicating wild genome sequences in the reference tomato genome. CONCLUSIONS: The methods developed in this work can be used to delineate genome introgressions, and subsequently contribute to development of molecular markers to aid phenotypic selection, fine mapping and discovery of candidate genes for important phenotypes, and for identification of novel variation for tomato improvement. These universal methods can easily be applied to other crop plants.

Evaluation of an activity monitor for the objective measurement of free-living physical activity in children with cerebral palsy.

OBJECTIVE: To explore the use of an activity monitor (AM) to objectively characterize free-living physical activity (F-LPA) in children with mobility impairment resulting from cerebral palsy (CP). DESIGN: First, a validation study compared outcomes from the AM with video evidence. Second, multiday F-LPA was characterized. Relationships between laboratory measures and F-LPA were explored. SETTING: The evaluation study was conducted in a laboratory environment. F-LPA monitoring was conducted in the participants' free-living environment. PARTICIPANTS: Convenience sample of ambulatory children (N=15; 11 boys, 4 girls) aged 5 to 17 years with CP undergoing gait analysis. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Accuracy of the AM for sitting/lying time, upright time, stepping time, and strides taken. Daily volumes of F-LPA of children with CP. RESULTS: AM outcomes in comparison with video-based analysis were (mean ± SD) 97.4%±2.7%, 101.1%±1.5%, 99.5%±6.6%, 105.6%±15.8%, and 103.8%±10.1% for sitting/lying time, upright time, standing time, stepping time, and stride count, respectively. Participants' daily F-LPA demonstrated considerable variation: mean standing time ± SD, 2.33±.96h/d; mean stepping time ± SD, 1.68±.86h/d; mean steps per day ± SD, 8477±4528; and mean sit-to-stand transitions per day ± SD, 76±49. Laboratory-measured cadence and mobility level were related to F-LPA, but not directly. CONCLUSIONS: The AM demonstrated excellent ability to determine sitting/lying and upright times in children with CP. Stepping time and stride count had lower levels of agreement with video-based analysis but were comparable to findings in previous studies. Crouch gait and toe walking had an adverse effect on outcomes. The F-LPA data provided additional information on children's performance not related to laboratory measures, demonstrating the added value of using this objective measurement technique.

Validity of method to quantify transtibial amputees' free-living prosthetic wearing times and physical activity levels when using suction suspension sockets.

Prostheses are prescribed to restore the mobility of people with amputated lower limbs. Monitoring the prosthesis wearing times and physical activity of prosthesis users would provide invaluable information regarding rehabilitation progress and suitability of the prosthesis. The validation of a method to determine wearing times and physical activity state, as well as strides taken, of amputees wearing suction suspension sockets is reported. Eight participants with transtibial amputation were fitted with custom-made suction sockets. Analysis algorithms were used to automatically characterize physical activity based on the pressure at the socket's relief valve. The algorithms were validated in a laboratory-based protocol that included walking, stair climbing, standing, sitting, donning, and doffing. Intraclass correlation coefficient (2,1) values of >0.98 were achieved with mean differences of - 2.0%, 0.3%, 1.3%, and 0.7% for agreement between "off," "static," and "dynamic" times and stride count, respectively, as determined by the analysis algorithms and a concurrent video analysis. This study demonstrates that an interpretation of the pressure at the pressure-relief valve of suction suspension sockets can be used to determine wearing times and activity state.

The effectiveness of behavioural interventions in the primary prevention of hepatitis C amongst injecting drug users: a randomised controlled trial and lessons learned.

AIM: To develop and evaluate the comparative effectiveness of behavioural interventions of enhanced prevention counselling (EPC) and simple educational counselling (SEC) in reducing hepatitis C viral (HCV) infection in sero-negative injecting drug users (IDU). DESIGN: Randomised controlled trial (RCT) of EPC intervention in comparison with simple educational counselling (SEC). SETTING SPECIALISED: Drug services in London and Surrey, United Kingdom. PARTICIPANTS AND MEASUREMENTS: Ninety five IDUs were recruited and randomised to receive EPC (n = 43) or SEC (n = 52). Subjects were assessed at baseline using the Addiction Severity Index (ASI), the Injecting Risk Questionnaire (IRQ), and Drug Injecting Confidence Questionnaire (DICQ). The primary outcome was measured by the rate of sero-conversion at 6 months and 12 months from baseline and by the ASI, IRQ and DICQ at 6 months from baseline. Hepatitis C testing was undertaken by the innovative test of the dried blood spot (DBS) test which increased the rate of testing by 4 fold compared to routine blood testing. FINDINGS SEVENTY: Eighty two subjects (82%) out of the 95 recruited were followed up at 6 months and 62 (65%) were followed up at 12 months. On the primary outcome measure of the rate of seroconversion, 8 out of 62 patients followed-up at twelve months seroconverted, three in the EPC group and five in the SEC group, indicating incidence rates of 9.1 per 100 person years for the EPC group, 17.2 per 100 person years for the SEC group, and 12.9 per 100 person years for the cohort as a whole. Analysis of the secondary outcome measures on alcohol use, risk behaviour, psychological measures, quality of life, showed no significant differences between the EPC and the SEC groups. However, there were significant changes on a number of measures from baseline values indicating positive change for both groups. CONCLUSION: We were not able to prove the efficacy of EPC in comparison with SEC in the prevention of hepatitis C in IDUs. This was related to low recruitment and retention rates of the participants. Moreover there was a low adherence rate to EPC. The study provided the benefits of developing and introducing behavioural interventions of the EPC and SEC and the DBS screening for Hepatitis C. Moreover the main lessons learnt were that piloting of a new intervention is a crucial first step before conducting pragmatic RCTs of psychological interventions in the field of addiction; that an infrastructure and culture for psychosocial interventions is needed to enable applied research in the service environment, and research funding is needed for enabling the recruitment of dedicated trained therapists for the delivery of these interventions.

Identification of Begomoviruses Infecting Crops and Weeds in Belize.

Plants including pepper, red kidney bean, squash, string bean and tomato, as well as weeds with viral symptoms were collected from five districts in Belize over a three year period with the aim of determining the diversity of the begomoviruses present. Sixty five percent of the samples screened via DNA hybridization produced signals indicative of begomovirus infection. Subsequent PCR amplifications and nucleotide sequence analyses revealed the presence of four begomoviruses in Belize. Pepper golden mosaic virus and Tomato mottle virus-[Flo] were found associated with tomato and sweet pepper and the former was also isolated from hot pepper. Merremia mosaic virus was found infecting hot pepper, sweet pepper and the weed species Euphorbia heterophylla. Euphorbia mosaic virus-[Yucatan Peninsula] was found in hot pepper and Euphorbia. This is the first report of the identification of begomoviruses in Belize.

Gastrointestinal symptoms in patients with end-stage renal disease undergoing treatment by hemodialysis or peritoneal dialysis.

OBJECTIVES: The aim of this study was to determine the prevalence of gastrointestinal symptoms in patients with end-stage renal disease undergoing hemodialysis (HD) or peritoneal dialysis (PD) treatment. METHODS: Patients undergoing HD or chronic ambulatory PD in the Department of Renal Medicine of our hospital were asked to complete a locally validated Rome II questionnaire. The same questionnaire was also administered to age- and gender-matched medical outpatients without renal failure, and community subjects. Patients on HD and their outpatient controls also completed the Hospital Anxiety and Depression Scale. RESULTS: A total of 148 patients with end-stage renal disease (HD 100, PD 48) completed the study. Their results were compared with those of 148 age- and gender-matched medical outpatients without renal failure and 148 randomly selected community controls. Patients on dialysis treatment were more likely to have abdominal pain, use laxatives, and report symptoms consistent with functional vomiting and irritable bowel syndrome (IBS) (70%, 55%, 16%, and 25%) than age- and gender-matched medical outpatients without renal failure (21%, 11%, 1%, and 6%) and community controls (16%, 4%, 0%, and 8%). Among HD patients and their outpatient controls, the differences appeared to be unrelated to anxiety or depression. CONCLUSIONS: Abdominal pain, laxative use, functional vomiting, and IBS were more common in uremic individuals than in controls without renal failure. Our data have not shown whether these differences are due to the uremic state itself, or, alternatively, its treatment.

Marker-Assisted Selection of Tomato Genotypes with the I-2 Gene for Resistance to Fusarium oxysporum f. sp. lycopersici Race 2.

Fusarium vascular wilt, caused by Fusarium oxysporum f. sp. lycopersici, affects tomatoes worldwide. Development of resistant varieties of tomato would constitute an economically and environmentally sound approach for the management of this disease. Resistance genes to F. oxysporum f. sp. lycopersici race 1 (I-1 gene) and race 2 (I-2 gene) were mapped to chromosome 11. The I-2 gene cluster includes one functional copy and six nonfunctional homologs of the I-2 gene. This report describes the design of primers based on the functional gene copy and the development of a multiplex polymerase chain reaction (PCR)-based method that has the ability to differentiate I-2 genotypes from genotypes without the I-2 gene. In these trials, 39 of the 40 genotypes tested with known reactions to race 2 gave the expected results. The only exception was the cultivar Plum Crimson carrying the I-3 gene for resistance, which confers resistance to F. oxysporum f. sp. lycopersici races 1, 2, and 3. This method was validated in three countries and by bioassays.

Gait impairments in a group of patients with incomplete spinal cord injury and their relevance regarding therapeutic approaches using functional electrical stimulation.

The purpose of this study is to determine the most important impairments affecting the gait pattern of the incomplete spinal cord injury (SCI) patient and the potential impact of their treatment. The study consists of two parts. Firstly, a survey amongst 16 professionals was done to find out the impact of the impairments in incomplete-SCI patients. Secondly, gait data from 21 individuals were analyzed to determine the most common impairments. Frequently observed and relevant impairments were: inadequate hip extension (occurrence 76%), limited hip flexion (52%), limited knee flexion (71%), excess of plantar flexion (76%), and impaired foot contact (52%). In conclusion, for gait improvement in incomplete spinal cord injured patients not only ankle movements must be treated, which is done frequently, but also hip extension/flexion and knee flexion are important for gait restoration. The impact on the gait and potential solutions of these impairments are discussed.

Immunodiagnosis of Prune dwarf virus using antiserum produced to its recombinant coat protein.

Certification represents the first line of defense against fruit tree viruses. For certification or surveys dealing with large number of samples, ELISA is still considered the technique of choice and requires a continuous supply of good quality antibodies. Prune dwarf virus (PDV) is among the major viruses affecting stone fruits; it belongs to the genus Ilarvirus named so for its isometric labile particles. Recombinant DNA technology was investigated for production of PDV antiserum to avoid labile virus purification and virus maintenance problems. The PDV coat protein gene (CP) was cloned into a protein expression bacterial plasmid vector which allowed a good level of expression of up to 2mg native protein/L culture. The recombinant PDV CP was injected into rabbits and the crude antiserum was successfully used in indirect ELISA at dilutions of up to 1:5000 to detect PDV in infected leaf samples. Similar results were obtained in dot blot immunoassays (DBIA). The antibodies were used in double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) and results were comparable to a reference commercial kit. The crude antiserum was efficiently used for coating ELISA plates, thereby reducing test costs.

Functional electric stimulation to augment partial weight-bearing supported treadmill training for patients with acute incomplete spinal cord injury: A pilot study.

OBJECTIVE: To study the application of partial weight-bearing (PWB) supported treadmill gait training augmented by functional electric stimulation (FES) in subjects with acute incomplete spinal cord injury (SCI). DESIGN: Before-after crossover trial with control (A) and intervention (B) periods. SETTING: Physiotherapy (PT) department of a spinal injuries unit in Scotland. PARTICIPANTS: Fourteen inpatients with acute incomplete SCI with American Spinal Injury Association class C or D injury. INTERVENTION: Training consisted of treadmill walking with PWB support augmented by FES. Subjects walked on the treadmill for up to 25 minutes a day, 5 days a week for 4 weeks. The intervention was compared with a 4-week control period in which standard PT was given. MAIN OUTCOME MEASURES: Overground walking endurance and speed, cadence, stride length, and observational gait analysis and walking speed, distance, and percentage PWB support on the treadmill. RESULTS: A greater increase in overground walking endurance was achieved after the intervention (AB group mean, 72.2m; confidence interval [CI], 39.8-104.6m; BA group mean, 63.8m; CI, -10.2 to 137.9m), as compared with after standard PT (AB group mean, 38.4m; CI, 1.8-75.0m; BA group mean, 60.1m; CI, 9.2-110.9m). A similar pattern was observed for overground walking speed. CONCLUSIONS: This pilot study indicated that PWB supported treadmill training with FES had a positive effect on overground gait parameters and could potentially accelerate gait training in subjects with incomplete SCI. A larger randomized trial is required to substantiate these findings.

A new method of using heart rate to represent energy expenditure: the Total Heart Beat Index.

OBJECTIVES: To develop a new method of representing energy expenditure using heart rate and to determine its reproducibility compared with the criterion standard of oxygen cost. DESIGN: Repeated-measures design. SETTING: University gait analysis laboratory and gymnasium at 2 spinal injury units. PARTICIPANTS: Twenty unimpaired adults and 17 subjects with spinal cord injury (SCI). INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Heart rate and oxygen consumption were measured on 20 unimpaired adults walking under controlled steady-state and nonsteady-state conditions. New methods of estimating energy expenditure by using heart rate were compared with oxygen consumption, oxygen cost, and Physiological Cost Index (PCI). Nine subjects with SCI, walking with and without functional electric stimulation, were assessed to determine use of these new measures with this group. Sensitivity to change of the new measurement techniques was investigated in 10 subjects with SCI, comparing wheelchair pushing to walking. RESULTS: The Total Heart Beat Index (THBI) was developed as a new index, calculated by dividing the total heartbeats during activity by distance traveled. High repeatability was found under steady-state and nonsteady-state conditions (intraclass correlation coefficients,.893-.995). Sensitivity to change in activity level was also shown. CONCLUSIONS: The THBI is a simple parameter to calculate from continuous heart rate data and provides a reproducible alternative to gas analysis and the PCI.

Tomato yellow leaf curl virus in the Dominican Republic: Characterization of an Infectious Clone, Virus Monitoring in Whiteflies, and Identification of Reservoir Hosts.

ABSTRACT Epidemics of tomato yellow leaf curl disease (TYLCD) in the Dominican Republic in the early to mid-1990s resulted in catastrophic losses to processing tomato production. As part of an integrated management approach to TYLCD, the complete nucleotide sequence of a full-length infectious clone of an isolate of Tomato yellow leaf curl virus (TYLCV) from the Dominican Republic (TYLCV-[DO]) was determined. The TYLCV-[DO] genome was nearly identical in sequence (>97%) and genome organization to TYLCV isolates from Israel and Cuba. This established that TYLCV-[DO] is a bonafide TYLCV isolate (rather than a recombinant virus, such as isolates from Israel [Mild], Portugal, Japan, and Iran), and provided further evidence for the introduction of the virus from the eastern Mediterranean. A reduction in the incidence of TYLCV in the northern and southern processing tomato production areas of the Dominican Republic has been associated with the implementation of a mandatory 3-month whitefly host-free period (including tomato, common bean, cucurbits, eggplant, and pepper). Monitoring TYLCV levels in whiteflies, by polymerase chain reaction with TYLCV-specific primers, established that the incidence of TYLCV decreased markedly during the host-free period, and then gradually increased during the tomato-growing season. In contrast, TYLCV persisted in whiteflies and tomato plants in an area in which the host-free period was not implemented. Surveys for TYLCV reservoir hosts, conducted to identify where TYLCV persists during the host-free period, revealed symptomless infections in a number of weed species. The implications of these findings for TYLCV management in the Dominican Republic are discussed.